AU5671899A - Collagen type i and type iii hemostatic compositions for use as a vascular sealant and wound dressing - Google Patents
Collagen type i and type iii hemostatic compositions for use as a vascular sealant and wound dressing Download PDFInfo
- Publication number
- AU5671899A AU5671899A AU56718/99A AU5671899A AU5671899A AU 5671899 A AU5671899 A AU 5671899A AU 56718/99 A AU56718/99 A AU 56718/99A AU 5671899 A AU5671899 A AU 5671899A AU 5671899 A AU5671899 A AU 5671899A
- Authority
- AU
- Australia
- Prior art keywords
- composition
- collagen
- collagen type
- type iii
- iii
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 239000000203 mixture Substances 0.000 title claims description 149
- 108010069502 Collagen Type III Proteins 0.000 title claims description 86
- 102000001187 Collagen Type III Human genes 0.000 title claims description 86
- 108010022452 Collagen Type I Proteins 0.000 title claims description 84
- 102000012422 Collagen Type I Human genes 0.000 title claims description 84
- 239000000565 sealant Substances 0.000 title claims description 63
- 229940096422 collagen type i Drugs 0.000 title claims description 59
- 230000002792 vascular Effects 0.000 title claims description 28
- 230000002439 hemostatic effect Effects 0.000 title description 21
- 102000008186 Collagen Human genes 0.000 claims description 115
- 108010035532 Collagen Proteins 0.000 claims description 115
- 229920001436 collagen Polymers 0.000 claims description 113
- 238000000034 method Methods 0.000 claims description 60
- 230000001070 adhesive effect Effects 0.000 claims description 51
- 239000000853 adhesive Substances 0.000 claims description 46
- 210000001519 tissue Anatomy 0.000 claims description 44
- 239000000178 monomer Substances 0.000 claims description 34
- 108010010803 Gelatin Proteins 0.000 claims description 27
- 229920000159 gelatin Polymers 0.000 claims description 27
- 235000019322 gelatine Nutrition 0.000 claims description 27
- 235000011852 gelatine desserts Nutrition 0.000 claims description 27
- 239000003795 chemical substances by application Substances 0.000 claims description 24
- 239000008273 gelatin Substances 0.000 claims description 24
- 108010049003 Fibrinogen Proteins 0.000 claims description 18
- 102000008946 Fibrinogen Human genes 0.000 claims description 18
- 230000008569 process Effects 0.000 claims description 18
- 108010073385 Fibrin Proteins 0.000 claims description 17
- 102000009123 Fibrin Human genes 0.000 claims description 17
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 claims description 17
- 229950003499 fibrin Drugs 0.000 claims description 17
- 229940012952 fibrinogen Drugs 0.000 claims description 17
- 238000004519 manufacturing process Methods 0.000 claims description 17
- 230000000379 polymerizing effect Effects 0.000 claims description 17
- 238000004132 cross linking Methods 0.000 claims description 15
- 108090000623 proteins and genes Proteins 0.000 claims description 15
- 108090000190 Thrombin Proteins 0.000 claims description 14
- 229960004072 thrombin Drugs 0.000 claims description 14
- 108010071289 Factor XIII Proteins 0.000 claims description 10
- 229940012444 factor xiii Drugs 0.000 claims description 9
- 229920001184 polypeptide Polymers 0.000 claims description 9
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 9
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 9
- 102000015225 Connective Tissue Growth Factor Human genes 0.000 claims description 8
- 108010039419 Connective Tissue Growth Factor Proteins 0.000 claims description 8
- 102000004079 Prolyl Hydroxylases Human genes 0.000 claims description 5
- 108010043005 Prolyl Hydroxylases Proteins 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 5
- 108010088751 Albumins Proteins 0.000 claims description 4
- 102000009027 Albumins Human genes 0.000 claims description 4
- 206010052428 Wound Diseases 0.000 description 36
- 208000027418 Wounds and injury Diseases 0.000 description 34
- 238000006116 polymerization reaction Methods 0.000 description 22
- 210000001367 artery Anatomy 0.000 description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 20
- 239000000515 collagen sponge Substances 0.000 description 18
- 241000283690 Bos taurus Species 0.000 description 17
- 239000000463 material Substances 0.000 description 17
- 208000032843 Hemorrhage Diseases 0.000 description 15
- 238000012360 testing method Methods 0.000 description 15
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 13
- 230000000740 bleeding effect Effects 0.000 description 13
- 238000007789 sealing Methods 0.000 description 13
- 239000003106 tissue adhesive Substances 0.000 description 12
- 239000000306 component Substances 0.000 description 11
- 229920000642 polymer Polymers 0.000 description 11
- 208000010110 spontaneous platelet aggregation Diseases 0.000 description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 10
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- 210000004027 cell Anatomy 0.000 description 10
- 241001465754 Metazoa Species 0.000 description 9
- 210000004204 blood vessel Anatomy 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 238000011282 treatment Methods 0.000 description 9
- 230000029663 wound healing Effects 0.000 description 9
- 210000004369 blood Anatomy 0.000 description 8
- 239000008280 blood Substances 0.000 description 8
- 230000023597 hemostasis Effects 0.000 description 8
- 210000000952 spleen Anatomy 0.000 description 8
- 238000001356 surgical procedure Methods 0.000 description 8
- 230000015572 biosynthetic process Effects 0.000 description 7
- 239000000975 dye Substances 0.000 description 7
- 239000000499 gel Substances 0.000 description 7
- 239000002874 hemostatic agent Substances 0.000 description 7
- 239000008188 pellet Substances 0.000 description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 230000009102 absorption Effects 0.000 description 6
- 238000010521 absorption reaction Methods 0.000 description 6
- 238000002583 angiography Methods 0.000 description 6
- 238000002399 angioplasty Methods 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 6
- 230000001965 increasing effect Effects 0.000 description 6
- 230000004048 modification Effects 0.000 description 6
- 238000012986 modification Methods 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 6
- 239000003431 cross linking reagent Substances 0.000 description 5
- 239000003814 drug Substances 0.000 description 5
- 210000001105 femoral artery Anatomy 0.000 description 5
- -1 for example Proteins 0.000 description 5
- 238000009472 formulation Methods 0.000 description 5
- 239000003102 growth factor Substances 0.000 description 5
- 230000035876 healing Effects 0.000 description 5
- 230000001976 improved effect Effects 0.000 description 5
- 238000011534 incubation Methods 0.000 description 5
- 208000014674 injury Diseases 0.000 description 5
- 102000004169 proteins and genes Human genes 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000009281 ultraviolet germicidal irradiation Methods 0.000 description 5
- 241000235058 Komagataella pastoris Species 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 239000003242 anti bacterial agent Substances 0.000 description 4
- 229940030225 antihemorrhagics Drugs 0.000 description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 4
- 239000012620 biological material Substances 0.000 description 4
- 238000005119 centrifugation Methods 0.000 description 4
- 229940038469 collagen hemostat Drugs 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 4
- 229940079593 drug Drugs 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 210000003128 head Anatomy 0.000 description 4
- 229940106780 human fibrinogen Drugs 0.000 description 4
- 239000001301 oxygen Substances 0.000 description 4
- 229910052760 oxygen Inorganic materials 0.000 description 4
- 210000002381 plasma Anatomy 0.000 description 4
- 210000004623 platelet-rich plasma Anatomy 0.000 description 4
- 238000010188 recombinant method Methods 0.000 description 4
- 150000003839 salts Chemical class 0.000 description 4
- 239000002002 slurry Substances 0.000 description 4
- 210000004872 soft tissue Anatomy 0.000 description 4
- 210000002435 tendon Anatomy 0.000 description 4
- 238000009736 wetting Methods 0.000 description 4
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 3
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 3
- 206010053567 Coagulopathies Diseases 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108010080379 Fibrin Tissue Adhesive Proteins 0.000 description 3
- 241000700159 Rattus Species 0.000 description 3
- 208000025865 Ulcer Diseases 0.000 description 3
- 238000004026 adhesive bonding Methods 0.000 description 3
- 239000000443 aerosol Substances 0.000 description 3
- 230000002776 aggregation Effects 0.000 description 3
- 238000004220 aggregation Methods 0.000 description 3
- 238000004873 anchoring Methods 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 229940096423 bovine collagen type i Drugs 0.000 description 3
- 239000001110 calcium chloride Substances 0.000 description 3
- 229910001628 calcium chloride Inorganic materials 0.000 description 3
- 229910001424 calcium ion Inorganic materials 0.000 description 3
- 230000035602 clotting Effects 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 238000002316 cosmetic surgery Methods 0.000 description 3
- 230000007812 deficiency Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 3
- 229910000397 disodium phosphate Inorganic materials 0.000 description 3
- 235000019800 disodium phosphate Nutrition 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 210000002950 fibroblast Anatomy 0.000 description 3
- 239000003292 glue Substances 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 230000028709 inflammatory response Effects 0.000 description 3
- 239000003999 initiator Substances 0.000 description 3
- 210000003734 kidney Anatomy 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 210000000056 organ Anatomy 0.000 description 3
- 230000001590 oxidative effect Effects 0.000 description 3
- 239000011546 protein dye Substances 0.000 description 3
- 230000004044 response Effects 0.000 description 3
- 229920001059 synthetic polymer Polymers 0.000 description 3
- 238000002560 therapeutic procedure Methods 0.000 description 3
- 229940075469 tissue adhesives Drugs 0.000 description 3
- 231100000397 ulcer Toxicity 0.000 description 3
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 description 2
- 108010039627 Aprotinin Proteins 0.000 description 2
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 2
- 102000000503 Collagen Type II Human genes 0.000 description 2
- 108010041390 Collagen Type II Proteins 0.000 description 2
- 208000005189 Embolism Diseases 0.000 description 2
- 102000016359 Fibronectins Human genes 0.000 description 2
- 108010067306 Fibronectins Proteins 0.000 description 2
- SXRSQZLOMIGNAQ-UHFFFAOYSA-N Glutaraldehyde Chemical compound O=CCCCC=O SXRSQZLOMIGNAQ-UHFFFAOYSA-N 0.000 description 2
- 206010018852 Haematoma Diseases 0.000 description 2
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 102000057297 Pepsin A Human genes 0.000 description 2
- 108090000284 Pepsin A Proteins 0.000 description 2
- 241000235648 Pichia Species 0.000 description 2
- 108010050808 Procollagen Proteins 0.000 description 2
- 208000002847 Surgical Wound Diseases 0.000 description 2
- 102000018594 Tumour necrosis factor Human genes 0.000 description 2
- 108050007852 Tumour necrosis factor Proteins 0.000 description 2
- 239000003146 anticoagulant agent Substances 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 229960004405 aprotinin Drugs 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000005540 biological transmission Effects 0.000 description 2
- 229910052791 calcium Inorganic materials 0.000 description 2
- 239000011575 calcium Substances 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 210000002808 connective tissue Anatomy 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 238000002405 diagnostic procedure Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 229940049370 fibrinolysis inhibitor Drugs 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000000834 fixative Substances 0.000 description 2
- 108020001507 fusion proteins Proteins 0.000 description 2
- 102000037865 fusion proteins Human genes 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 229960002897 heparin Drugs 0.000 description 2
- 229920000669 heparin Polymers 0.000 description 2
- 238000000265 homogenisation Methods 0.000 description 2
- 229920002674 hyaluronan Polymers 0.000 description 2
- 229960003160 hyaluronic acid Drugs 0.000 description 2
- 239000005457 ice water Substances 0.000 description 2
- 238000002513 implantation Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- ZPNFWUPYTFPOJU-LPYSRVMUSA-N iniprol Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@H]2CSSC[C@H]3C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC=4C=CC=CC=4)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=4C=CC=CC=4)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC2=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]2N(CCC2)C(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N2[C@@H](CCC2)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N2[C@@H](CCC2)C(=O)N3)C(=O)NCC(=O)NCC(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N1)C(C)C)[C@@H](C)O)[C@@H](C)CC)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 ZPNFWUPYTFPOJU-LPYSRVMUSA-N 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002350 laparotomy Methods 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000011587 new zealand white rabbit Methods 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 239000001254 oxidized starch Substances 0.000 description 2
- 235000013808 oxidized starch Nutrition 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 230000035515 penetration Effects 0.000 description 2
- 229940111202 pepsin Drugs 0.000 description 2
- KHIWWQKSHDUIBK-UHFFFAOYSA-N periodic acid Chemical compound OI(=O)(=O)=O KHIWWQKSHDUIBK-UHFFFAOYSA-N 0.000 description 2
- 238000007539 photo-oxidation reaction Methods 0.000 description 2
- 230000004962 physiological condition Effects 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 229910000679 solder Inorganic materials 0.000 description 2
- 229960003766 thrombin (human) Drugs 0.000 description 2
- LCPVQAHEFVXVKT-UHFFFAOYSA-N 2-(2,4-difluorophenoxy)pyridin-3-amine Chemical compound NC1=CC=CN=C1OC1=CC=C(F)C=C1F LCPVQAHEFVXVKT-UHFFFAOYSA-N 0.000 description 1
- FDSYTWVNUJTPMA-UHFFFAOYSA-N 2-[3,9-bis(carboxymethyl)-3,6,9,15-tetrazabicyclo[9.3.1]pentadeca-1(15),11,13-trien-6-yl]acetic acid Chemical compound C1N(CC(O)=O)CCN(CC(=O)O)CCN(CC(O)=O)CC2=CC=CC1=N2 FDSYTWVNUJTPMA-UHFFFAOYSA-N 0.000 description 1
- SLXKOJJOQWFEFD-UHFFFAOYSA-N 6-aminohexanoic acid Chemical compound NCCCCCC(O)=O SLXKOJJOQWFEFD-UHFFFAOYSA-N 0.000 description 1
- 102100033312 Alpha-2-macroglobulin Human genes 0.000 description 1
- BSYNRYMUTXBXSQ-UHFFFAOYSA-N Aspirin Chemical compound CC(=O)OC1=CC=CC=C1C(O)=O BSYNRYMUTXBXSQ-UHFFFAOYSA-N 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 102000004506 Blood Proteins Human genes 0.000 description 1
- 108010017384 Blood Proteins Proteins 0.000 description 1
- 206010006797 Burns first degree Diseases 0.000 description 1
- 206010006802 Burns second degree Diseases 0.000 description 1
- 206010006803 Burns third degree Diseases 0.000 description 1
- 102000030523 Catechol oxidase Human genes 0.000 description 1
- 108010031396 Catechol oxidase Proteins 0.000 description 1
- 206010052114 Conjunctival bleb Diseases 0.000 description 1
- 208000028006 Corneal injury Diseases 0.000 description 1
- 229920001651 Cyanoacrylate Polymers 0.000 description 1
- GUBGYTABKSRVRQ-WFVLMXAXSA-N DEAE-cellulose Chemical compound OC1C(O)C(O)C(CO)O[C@H]1O[C@@H]1C(CO)OC(O)C(O)C1O GUBGYTABKSRVRQ-WFVLMXAXSA-N 0.000 description 1
- 206010015150 Erythema Diseases 0.000 description 1
- 108010088842 Fibrinolysin Proteins 0.000 description 1
- 206010016717 Fistula Diseases 0.000 description 1
- 206010018691 Granuloma Diseases 0.000 description 1
- 241000700721 Hepatitis B virus Species 0.000 description 1
- 208000034693 Laceration Diseases 0.000 description 1
- PHSRRHGYXQCRPU-AWEZNQCLSA-N N-(3-oxododecanoyl)-L-homoserine lactone Chemical compound CCCCCCCCCC(=O)CC(=O)N[C@H]1CCOC1=O PHSRRHGYXQCRPU-AWEZNQCLSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010029113 Neovascularisation Diseases 0.000 description 1
- 208000005890 Neuroma Diseases 0.000 description 1
- 101100401106 Neurospora crassa (strain ATCC 24698 / 74-OR23-1A / CBS 708.71 / DSM 1257 / FGSC 987) met-7 gene Proteins 0.000 description 1
- 206010030113 Oedema Diseases 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 229940122791 Plasmin inhibitor Drugs 0.000 description 1
- 102000013566 Plasminogen Human genes 0.000 description 1
- 108010051456 Plasminogen Proteins 0.000 description 1
- 102000004179 Plasminogen Activator Inhibitor 2 Human genes 0.000 description 1
- 108090000614 Plasminogen Activator Inhibitor 2 Proteins 0.000 description 1
- 229920001744 Polyaldehyde Polymers 0.000 description 1
- 108010015078 Pregnancy-Associated alpha 2-Macroglobulins Proteins 0.000 description 1
- 208000004210 Pressure Ulcer Diseases 0.000 description 1
- 102000016611 Proteoglycans Human genes 0.000 description 1
- 108010067787 Proteoglycans Proteins 0.000 description 1
- 206010061481 Renal injury Diseases 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 1
- 239000004826 Synthetic adhesive Substances 0.000 description 1
- 208000031737 Tissue Adhesions Diseases 0.000 description 1
- 108060008539 Transglutaminase Proteins 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 208000018756 Variant Creutzfeldt-Jakob disease Diseases 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 108010006886 Vitrogen Proteins 0.000 description 1
- 238000012084 abdominal surgery Methods 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 229960001138 acetylsalicylic acid Drugs 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000012190 activator Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000013466 adhesive and sealant Substances 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 102000003801 alpha-2-Antiplasmin Human genes 0.000 description 1
- 108090000183 alpha-2-Antiplasmin Proteins 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 229960002684 aminocaproic acid Drugs 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 229940127090 anticoagulant agent Drugs 0.000 description 1
- 229940127219 anticoagulant drug Drugs 0.000 description 1
- 229910001422 barium ion Inorganic materials 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 239000012503 blood component Substances 0.000 description 1
- 210000001124 body fluid Anatomy 0.000 description 1
- 239000010839 body fluid Substances 0.000 description 1
- 208000005881 bovine spongiform encephalopathy Diseases 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229960005069 calcium Drugs 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000012292 cell migration Effects 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000017455 cell-cell adhesion Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000013043 chemical agent Substances 0.000 description 1
- 238000007385 chemical modification Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000000038 chest Anatomy 0.000 description 1
- 230000015271 coagulation Effects 0.000 description 1
- 238000005345 coagulation Methods 0.000 description 1
- 230000037319 collagen production Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 210000004087 cornea Anatomy 0.000 description 1
- 238000002586 coronary angiography Methods 0.000 description 1
- 238000007887 coronary angioplasty Methods 0.000 description 1
- 208000029078 coronary artery disease Diseases 0.000 description 1
- 210000004351 coronary vessel Anatomy 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 229940072645 coumadin Drugs 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 238000009295 crossflow filtration Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- NLCKLZIHJQEMCU-UHFFFAOYSA-N cyano prop-2-enoate Chemical class C=CC(=O)OC#N NLCKLZIHJQEMCU-UHFFFAOYSA-N 0.000 description 1
- 239000000824 cytostatic agent Substances 0.000 description 1
- 230000001085 cytostatic effect Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002939 deleterious effect Effects 0.000 description 1
- 238000009795 derivation Methods 0.000 description 1
- 239000003599 detergent Substances 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000002183 duodenal effect Effects 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 231100000321 erythema Toxicity 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000001815 facial effect Effects 0.000 description 1
- 229960002089 ferrous chloride Drugs 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 230000035557 fibrillogenesis Effects 0.000 description 1
- 239000003527 fibrinolytic agent Substances 0.000 description 1
- 230000003176 fibrotic effect Effects 0.000 description 1
- 230000003890 fistula Effects 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 238000007429 general method Methods 0.000 description 1
- 238000002682 general surgery Methods 0.000 description 1
- 210000004013 groin Anatomy 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 229940005809 human factor xiii Drugs 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 230000009851 immunogenic response Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000010348 incorporation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 239000012678 infectious agent Substances 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- NMCUIPGRVMDVDB-UHFFFAOYSA-L iron dichloride Chemical compound Cl[Fe]Cl NMCUIPGRVMDVDB-UHFFFAOYSA-L 0.000 description 1
- 208000037806 kidney injury Diseases 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000002406 microsurgery Methods 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 210000004165 myocardium Anatomy 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 230000000399 orthopedic effect Effects 0.000 description 1
- 239000007800 oxidant agent Substances 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 210000003200 peritoneal cavity Anatomy 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 239000008177 pharmaceutical agent Substances 0.000 description 1
- 239000008194 pharmaceutical composition Substances 0.000 description 1
- 239000008363 phosphate buffer Substances 0.000 description 1
- 230000036470 plasma concentration Effects 0.000 description 1
- 229940012957 plasmin Drugs 0.000 description 1
- 239000002806 plasmin inhibitor Substances 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 239000002985 plastic film Substances 0.000 description 1
- 229920006255 plastic film Polymers 0.000 description 1
- 229920000058 polyacrylate Polymers 0.000 description 1
- 239000003505 polymerization initiator Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000001323 posttranslational effect Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 239000003380 propellant Substances 0.000 description 1
- 108020001775 protein parts Proteins 0.000 description 1
- 229940024999 proteolytic enzymes for treatment of wounds and ulcers Drugs 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000009103 reabsorption Effects 0.000 description 1
- 230000035484 reaction time Effects 0.000 description 1
- 230000001172 regenerating effect Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000009877 rendering Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 231100000241 scar Toxicity 0.000 description 1
- 239000003566 sealing material Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 1
- CHQMHPLRPQMAMX-UHFFFAOYSA-L sodium persulfate Substances [Na+].[Na+].[O-]S(=O)(=O)OOS([O-])(=O)=O CHQMHPLRPQMAMX-UHFFFAOYSA-L 0.000 description 1
- AKHNMLFCWUSKQB-UHFFFAOYSA-L sodium thiosulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=S AKHNMLFCWUSKQB-UHFFFAOYSA-L 0.000 description 1
- 235000019345 sodium thiosulphate Nutrition 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 125000001424 substituent group Chemical group 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000003685 thermal hair damage Effects 0.000 description 1
- 229960000103 thrombolytic agent Drugs 0.000 description 1
- 238000007483 tonsillectomy Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- QQJLHRRUATVHED-UHFFFAOYSA-N tramazoline Chemical compound N1CCN=C1NC1=CC=CC2=C1CCCC2 QQJLHRRUATVHED-UHFFFAOYSA-N 0.000 description 1
- 229960001262 tramazoline Drugs 0.000 description 1
- GYDJEQRTZSCIOI-LJGSYFOKSA-N tranexamic acid Chemical compound NC[C@H]1CC[C@H](C(O)=O)CC1 GYDJEQRTZSCIOI-LJGSYFOKSA-N 0.000 description 1
- 229960000401 tranexamic acid Drugs 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 102000003601 transglutaminase Human genes 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 239000002753 trypsin inhibitor Substances 0.000 description 1
- 210000003454 tympanic membrane Anatomy 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- 238000007631 vascular surgery Methods 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- PJVWKTKQMONHTI-UHFFFAOYSA-N warfarin Chemical compound OC=1C2=CC=CC=C2OC(=O)C=1C(CC(=O)C)C1=CC=CC=C1 PJVWKTKQMONHTI-UHFFFAOYSA-N 0.000 description 1
- 238000003466 welding Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/10—Polypeptides; Proteins
- A61L24/102—Collagen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/32—Proteins, polypeptides; Degradation products or derivatives thereof, e.g. albumin, collagen, fibrin, gelatin
- A61L15/325—Collagen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/043—Mixtures of macromolecular materials
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/10—Polypeptides; Proteins
- A61L24/104—Gelatin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B17/00—Surgical instruments, devices or methods, e.g. tourniquets
- A61B17/00491—Surgical glue applicators
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B17/00—Surgical instruments, devices or methods, e.g. tourniquets
- A61B17/0057—Implements for plugging an opening in the wall of a hollow or tubular organ, e.g. for sealing a vessel puncture or closing a cardiac septal defect
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Surgery (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Toxicology (AREA)
- Dermatology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Hematology (AREA)
- Materials Engineering (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Pharmacology & Pharmacy (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Materials For Medical Uses (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Description
WO 00/09018 PCTIUS99/18095 5 COLLAGEN TYPE I AND TYPE III HEMOSTATIC COMPOSITIONS FOR USE AS A VASCULAR SEALANT AND WOUND DRESSING 10 CROSS REFERENCE TO RELATED APPLICATIONS This application is a continuation-in-part of U.S. application provisional no. 15 60/095,977 FIELD OF THE INVENTION The present invention is directed to polymerized recombinant type I and/or type III collagen-based compositions and combinations thereof, including gelatin-based 20 compositions, for medical use as sealants and wound dressings. The present invention is further directed to the preparation of such compositions. These compositions are useful as sealants in a variety of medical applications, including vascular plug type devices, wound closure devices, tendon wraps for preventing the formation of adhesion following surgical procedures, and dressings for use to treat incisions, seeping wounds, and the like, 25 and as medical adhesives for bonding tissues. In a further aspect of the present invention, the compositions include agents which induce wound healing or provide additional beneficial characteristics desired in a tissue sealant. More particularly, the compositions of the present invention are can be used as vascular sealants. 30 BACKGROUND OF THE INVENTION Mechanical, Chemical, Synthetic and Autologous Adhesion Techniques. The ability to bond biological tissues is an important area of investigation for biomedical researchers. Attempts to provide desired adhesion through purely mechanical bonding have proven to be neither convenient nor permanent. (Buonocore, M., Adhesion in 35 Biological Systems, R. S. Manly, ed., Academic Press, New York, 1970, Chap. 15.) For example, the conventional methods of choice to close incisions in soft tissue following WO 00/09018 PCT/US99/18095 2 5 surgery, injury, and the like have been sutures and staples. These techniques and methods are limited by, for example, tissue incompatibility with sutures or staples which may cause painful and difficult to treat fistulas granulomas and neuromas. Mechanical means can also be limited to being purely adhesive, and thus not fully satisfactory as compared to sealants, applied to close wounds that are bleeding or seeping, etc. Sutures 10. and staples may also tend to cut through weak parenchymatous or poorly vascularized tissue. Sutures can leave behind a tract which can allow for leakage of fluids and organisms. Sutures can be further problematic in that the needle for any suture is larger than the thread attached to it and the needle tract is thus larger than can be filled by the thread used to form the sutures. 15 In addition, limits are imposed by the manual and visual dexterity required on the part of the surgeon and the excessive amount of time needed for the use of sutures or staples in microsurgeries. Furthermore, the joints in the gaps between staples or sutures, even when properly applied, the staples or sutures are inherently weak or may structurally weaken over time and leak. 20 Several investigators have worked on laser closure of wounds. (See, e.g., Abergel, R.P. et al. (1986) J. Am. Acad. Dermatol. 14(5):810-814; Cespanyi, E. et al. (1987) J. Surg. Res. 42(2):147-152; Oz, M.C. et al. (1989) Lasers Surg. Med. 9(3):248 253; Oz, M.C. et al. (1991) Am. Surg. 57(5):275-279; and Oz, M.C. et al. (1993) J. Clin. Laser Med. Surg. 11(3):123-126.) Early efforts concentrated on welding tissues using 25 lasers of different wavelengths applied directly to wound edges and investigating the microstructural basis of the tissue fusion thus produced. Researchers proposed that a homogenizing change in collagen with interdigitation of altered individual fibrils. (See, e.g., Schober, R. et al. (1986) Science 232(4756):1421-1422.) Investigators explored the idea of heating the collagen fibrils above a threshold level allowed for cross-linking. 30 However, the heat necessary to allow this reaction causes collateral thermal damage. This is undesirable as even a slight distortion in, for example, ocular tissue may have functional consequences. Also, in the event of laser weld failure, the edges of the tissues may be damaged by the original treatment and cannot be re-exposed to laser energy.
WO 00/09018 PCT/US99/18095 3 5 Further research attempted to enhance heat-activated cross-linking by placing a dye in the wound. It was reported that matching the absorbance of the dye with the laser wavelength achieved an adhesive effect with less laser power output and collateral thermal injury. (See, e.g., Chuck, R.S. et al. (1989) Lasers Surg. Med. 9(5):471-477; and Oz, M.C. et al. (1990) J. Vasc. Surg. 11(5):718-725.) Coupling the dye with a protein to 10 create a tissue "solder" was also investigated. The protein commonly used is fibrinogen, and, in particular, autologous fibrinogen, which is used to avoid problems of the transfer of viral diseases through the use of blood components from pool donors. In previous applications, fibrinogen has been obtained as a fraction of whole blood, contains other blood elements, such as clotting factors. Application of such a protein-dye mixture in 15 various animal models proved to be an improvement to dye alone. (See, e.g., Moazami, N. et al. (1990) Arch. Surg. 125(11):1452-1454; and Oz et al. (1990).) However, direct application in humans was prevented due to the need to isolate the necessary protein fibrinogen from the patient prior to the procedure to avoid risks of infection from donor plasma. Other proteins, for example, albumin, were unsatisfactory substitutes as welds of 20 comparable strength were not achieved. Comparisons of protein-dye applications and sutured closures show that the protein-dye applications produce less of an inflammatory response and result in greater collagen production, greater mean peak stress at rupture, and better cosmesis. (See, e.g., Wider, T.M. et al. (1991) Plast. Reconstr. Surg. 88(6):1018-1025.) Ophthalmologic 25 applications of such a tissue solder have included the sealing of conjunctival blebs, sclerostomy, closure of retinectomies, and thermokeratoplasty. (See, e.g., Fink, A.J. et al. (1986) Am. J. Ophthalmol. 101(6):695-699; and Latina, M.A. et al. (1990) Arch. Ohthalmol. 108(12):1745-1750.) Due to the deficiencies and limitations of mechanical means, such as the above 30 mentioned sutures, staples, and laser techniques, efforts were made to develop synthetic polymers, such as, for example, cyanoacrylates, as biomedical adhesives and sealants. These plastic materials, however, induce inflammatory tissue reactions. In addition, the ability of these materials to establish permanent bonding under physiological conditions has yet to be fully realized.
WO 00/09018 PCT/US99/18095 4 5 The known toxicity associated with synthetic adhesives has led to investigators to the development of biologically-derived adhesives as bonding materials. Fibrin-based glues, for example, have commanded considerable attention. (See, e.g., Epstein, G. H. et al. Ann. Otol. Rhinol. Laryngol. 95: 40-45 (1986); Kram, H. B et al. Arch. Surg. 119: 1309-1311 (1984); Scheele, J. et al. Surgery 95: 6-12 (January 1984); and Siedentop, K. 10 H. et al. Laryngoscooe 93: 1310-1313 (1983) for general discussion of fibrin adhesives.) Commercial fibrin tissue adhesives are derived from human plasma and thus pose potential health risks such as adverse immunogenic reactions and transmission of infectious agents, such as, for example, Hepatitis B virus. Moreover, the bond strength imparted by such adhesives are relatively weak compared to collagen adhesives. (See, for 1 5 example, De Toledo, A. R. et al. Assoc. for Res. in Vision and Ophthalmology, Annual Meeting Abstract, Vol. 31, 317 (1990).) Accordingly, there is a need for safe and effective biologically compatible tissue adhesives for biomedical applications. More recently, combination products have been devised for use as tissue adhesives and sealants. The use of a combination of three separately prepared substances, 20 human fibrinogen cryoprecipitate, thrombin in the presence of calcium ion, and Factor XIII concentrate, to obtain a glue for application in skin graft applications, myringoplasty, repair of dural defects, hemeostatis after tonsillectomy, and tracheoplasty has been described. (See, Staindl (Ann. Otol (1979) 88:413-418).) In this same time frame, Immuno-AG, Vienna, Austria, began producing and commercializing a two 25 component "fibrin seal" system, wherein one component contains highly concentrated human fibrinogen, Factor XIII, and other human plasma protein, prepared from pooled blood, and the other component supplies thrombin and calcium ion. The two components are added together in the presence of a fibrinolysis inhibitor. After application, coagulation and fibrin cross-linking occur. Eventually, the seal may lyse in the process of 30 healing of the wound or trauma which accompanies the reconstruction of the tissue. The development of an applicator device for this system which mixes and applies the two components of the system simultaneously has been described. (Redl, H., et al., "Biomaterials 1980," Winter, G. D., et al., eds. (1982), John Wiley & Sons, Ltd., at page 669-675.) These combination systems and their uses have been described widely. (See, WO 00/09018 PCT/US99/18095 5 5 e.g., Seelich, T., J Head and Neck Pathol (1982) 3:65-69; O'Connor, A. F., et al., Otolaryngol Head Neck Surg (1982) 90:347-348; Marquet, J., J Head and Neck Pathol (1982) 3:71-72; Thorson, G. K., et al., J Surg Oncol (1983) 24:221-223.) It has also been reported that the addition of barium ion to this fibrin glue system in the treatment of a bleeding duodenal sinus facilitates follow-up surveillance. (See, for example, McCarthy, 10 P. M., et al., Mayo Clin Pros (1987) 62:317-319; Portmann M., J Head and Neck Pathol (1982) 3:96; Panis, R., ibid., 94-95.) Efforts have recently focused on methods which seek to avoid health issues raised by the use of blood plasma-derived products in commercially available tissue adhesive products and systems. Attempts have been made to isolate an autologous counterpart of 15 the fibrinogen-containing component. (See, for example, Feldman, M. C., et al., Arch Otolaryngol-Head and Neck Surg (1988) 114:182-185; Feldman, M. C., et al., Arch Ophthalmol (1987) 105:963-967; Feldman, M. C., et. al., M J Otolog (1988) 9:302-305; Silberstein, L. E., et al., Transfusion (1988) 28:319-321.) Use of autologous fibrinogen preparations also have obvious limitations. 20 Vascular Sealants. One critical aspect of tissue adhesion is the sealing of wounds, and, in particular, vascular punctures and other vascular wounds resulting from, for example, surgery. For example, percutaneously accessing major vascular structures is a key step in a variety of diagnostic and therapeutic procedures, including Percutaneous Transluminal 25 Coronary Angioplasty (PTCA), Percutaneous Coronary Angiography, and Percutaneous Coronary Atherectomy. In a percutaneous intravascular procedure, access to the vascular space is generally obtained using the so-called Seldinger technique where, first, a hollow needle is used to create a puncture wound through the skin, the underlying muscle tissue, and the wall of a selected blood vessel, such as the femoral artery. Next, a guidewire is 30 inserted through the tubular needle until its distal end is located in the blood vessel, at which time the needle is stripped off of the guidewire and replaced with an introducer sheath and dilator. The introducer sheath typically includes a self-sealing hemostatic valve on its proximal end for sealing around the guidewire. The guidewire is then advanced into the vascular space through the introducer and directed to a preselected area WO 00/09018 PCT/US99/18095 6 5 of the vascular system. Once the guidewire is positioned, a catheter is advanced over the guidewire to the desired area. Once the procedure has been completed and the catheter and the introducer sheath are removed from the puncture site, there may be profuse bleeding, especially when the patient has been on anticoagulant therapy such as heparin, coumadin, aspirin, or 10 thrombolytic agents. The most common method used to prevent post-procedure bleeding at the access site involves the application of direct pressure to the perforation site until normal physiologic pathways have sealed the access site. There are several problems with this method. First, the pressure application technique may fail to prevent hemorrhage. Such a hemorrhage may be life-threatening or can lead to a large 1 5 hematoma. A large hematoma in the groin, for instance, may compromise the major nerve supply to the anterior lower extremity. Secondly, the pressure application technique extends the length of the in-hospital stay. For example, a PTCA may be completed in 2 to 3 hours, but the patient will typically be hospitalized for several additional hours or overnight to allow the access site 20 to seal physiologically. During this extended hospital stay the patient is required to stay immobile, often with a sand bag taped to the patient's thigh, such as in the case of femoral artery access. These and other complications are exacerbated where PTCA procedures are performed in elderly patients who commonly have arteries with reduced natural elasticity. 25 The access perforation in a relatively inelastic artery does not contract or shrink upon itself to the same extent as in an artery of normal elasticity. The resulting undeflected perforation is typically two to three times larger than an access perforation in a normal artery, further complicating the initiation of hemostasis and the normal physiologic sealing of the access site. 30 More than 500,000 PTCAs were performed worldwide in 1992 (Cowen Report, March 1993), and several times that number of other procedures requiring accessing major vascular structures percutaneously and were also performed. Thus, the increased length of in-hospital stay necessitated by the pressure application technique considerably increases the expense of procedures requiring such vascular access.
WO 00/09018 PCT/US99/18095 7 5 A technique that would allow faster and safer sealing of a vascular access site would save a significant amount of health care resources. Medical literature has addressed the problem of achieving hemostasis following removal of a percutaneously applied intravascular introducer in such uses as angiography or angioplasty by a number of divergent means. U.S. Patent No. 5,290,310 describes a device for delivering a 10 collagen plug subcutaneously against a penetration site in a wall of a blood vessel. An instrument containing a toroidal-shaped collagen plug within a barrel thereof is made to surround the exterior of a tubular introducer. The instrument includes a pusher mechanism for ejecting the collagen plug into the puncture wound and against the exterior wall of the blood vessel at the site of the puncture. This device relies upon a 15 collagen plug which is derived from animal sources and is therefore comprised primarily of heterotrimer collagen type I. U.S. Patent No. 5,129,882 also discloses a surgical implement for injecting a hemostatic agent in a puncture wound by routing the injection device through the lumen of the introducer sheath after it has been retracted sufficiently so that the distal end 20 thereof is no longer in the blood vessel. Deploying a plunger, the hemostatic agent is forced out of the instrument and against the exterior wall of the artery proximate the puncture wound. U.S. Patent Nos. 4,744,364, 4,852,974, 4,890,612, 5,021,059, and 5,222,974 each describe a method and apparatus for effecting hemostasis by inserting an anchoring 25 device through the puncture wound and into the blood vessel while using a filament attached to the anchoring device to inject an appropriate sealant into the wound. The anchoring device prevents entrance of the sealing material into the blood vessel and serves as an anchor and guide for addressing selected vessels. Still other devices for injecting a hemostatic agent into a puncture wound 30 following a vascular procedure are described in U.S. Patent Nos. 5,281,197, 4,838,280, 5,192,300, and 4,738,658 and in published European Patent Application 0 476 178A1. Collagen/Gelatin As A Biomaterial. Collagen, the major connective tissue protein in animals, possesses numerous characteristics not seen in synthetic polymers. Characteristics of collagen include good compatibility with living tissue, WO 00/09018 PCT/US99/18095 8 5 promotion of cell growth, and absorption and assimilation of implantations. (See, e.g., Shimizu, R. et Al. Biomat. Med. Dev. Art. Org., 5(1): 49-66 (1977).) These same characteristics are also true of gelatins, derivation products of collagens. Various applications of collagen as a biomaterial are being tested, for example, the use of collagens in dialysis membranes of artificial kidney, artificial cornea, vitreous 10 body, artificial skin and blood vessels, hemostatic agents, soft contact lesnes, and in surgery. (Sterzel, K. H. et al. Ameri. Soc. Artif. Int. Organs 17: 293 (1971), Rubin, A. L. et al. Nature 230: 120 (1971), and U.S. Pat. No. 4,581,030, Dunn, M. et al. Amer. Soc. Artif. Int. Organs 17: 421 (1971), Krajicek, M. et al. J. Surg. Res. 4, 290 (1964), U.S. Pat. No. 4,215,200, U.S. Pat. Nos. 4,264,155; 4,264,493; 4,349,470; 4,388,428; 4,452,925, 15 and 4,650,616, and Chvapil, M. et al. Int. Rev. Conn. Tiss. Res. 6: 1-61 (1973).) Natural collagen fibers, however, are basically insoluble in mature tissues because of covalent intermolecular cross-links that convert collagen into an infinite cross-linked network. Dispersal and solubilization of native collagen can be achieved by treatment with various proteolytic enzymes which disrupt the intermolecular bonds and remove 20 immunogenic non-helical end regions without affecting the basic rigid triple-helical structure which imparts the desired characteristics of collagen. (See, e.g., U.S. Pat. Nos. 3,934,852; 3,121,049; 3,131,130; 3,314,861; 3,530,037; 3,949,073; 4,233,360, and 4,488,911 for general methods for preparing purified soluble collagen.) Various methods and materials have been proposed for modifying collagen to 25 render it more suitable as biomedical adhesives. (See, e.g., De Toledo, A. R. et al. Assoc. for Res. in Vision and Ophthalmology, Annual Meeting Abstract, Vol. 31, 317 (1990); Lloyd et al., "Covalent Bonding of Collagen and Acrylic Polymers," American Chemical Society Symposium on Biomedical and Dental Applications of Polymers, Polymer Science and Technology, Vol. 14, Plenum Press (Gebelein and Koblitz eds.), New York, 30 1980, pp. 59-84; Shimizu et al., Biomat. Med. Dev. Art. Org., 5(1): 49-66 (1977); and Shimizu et al., Biomat. Med. Dev. Art. Org., 6(4): 375-391 (1978), for general discussion on collagen and synthetic polymers.) In many instances, the prior modified collagen based adhesives suffer from various deficiencies, including (1) cross linking/polymerization reactions that generate exothermic heat, (2) long reaction times, WO 00/09018 PCT/US99/18095 9 5 and (3) reactions that are inoperative in the presence of oxygen and physiological pH ranges. (See, eg., Lee M. L. et al. Adhesion in Biological Systems, R. S. Manly, ed., Academic Press, New York, 1970, Chap. 17.) Moreover, many of the prior modified collagen-based adhesives contain toxic materials, rendering them unsuitable for biomedical use. (See, for example, Buonocore, M. G. (1970) and U.S. Pat. No. 10 3,453,222.) Additionally, the use of collagen-based adhesives also presents immunological concerns as such adhesives have been derived from animal sources and typically bovine sources. Studies with respect to the use of such collagens as injectible devices have reported minor inflammatory responses. More recently, potential issues regarding the 1 5 transmission of disorders to humans related to bovine spongiform encephalopathy ("mad cow disease") have focused attention, especially in Europe, to limiting the use of animal, and particularly, bovine-sourced materials. Notwithstanding these deficiencies, certain collagen-based adhesives, reportedly having appropriate adhesive strength and utility in many medical applications, 20 particularly involving soft tissues, have been described. (See, e.g., U.S. Patent Nos. 5,219,895, 5,614,587, 5,582,834, 5,575,997, 5,354,336, and 4,600,574.) The literature identify the use of type I and type II in collagen-based adhesives wherein purified collagen types I and II are chemically modified to form monomers soluble at physiological conditions and polymerized to form compositions having adhesive and 25 sealant properties. Notably, the reports are limited to collagen-based adhesives composed of collagens derived from natural sources which represent a collagen mixture. For example, type I collagen as isolated from natural sources typically contains approximately 10-20% type III and other collagens, depending upon the tissue source used, and about 90-80% type I collagen. With respect to the "collagen type I" mixtures, 30 the literature further teaches only the use of collagen as an adhesive as a consequence of its structural characteristics, or, alternatively, the use of predominantly collagen type I heterotrimers, as compared to collagen type I homotrimers which have been implicated in the epithelial cell attachment. (See, e.g., Ghersi, et al., 1989, Eur. J. Cell Biol. 50:279 84)).
WO 00/09018 PCTIUS99/18095 10 5 Available reports do not refer to collagen type III, the unexpected hemostatic characteristics of type III collagen, or the use of recombinant collagens which would allow the first chemical modification step, as described in the art, to be avoided. In summary, there is a need in the art for compositions useful as sealants and wound dressings that permit faster and safer healing, that minimize the risks of infection 10 from donor, including non-human, sources, that increase convenience and permanence, that minimize demands on surgical resources and time, and that demonstrate superior biocompatibility. In addition, there is a need for biologically-derived adhesives that offer improved convenience and permanence over currently available formulations, and that promote less-invasive treatment, resulting in improved patient comfort and shorter time 15 under medical supervision. Additionally, such compositions would preferably offer improved bond strength. There is also a need for non-adhesive compositions that provide the above-named advantages of safer and more effective healing and that are biologically compatible. 20 SUMMARY OF THE INVENTION The present invention includes biologically compatible, collagen type III and/or type I products with sealant properties which can be formed using soluble recombinantly derived collagen type III and/or type I monomers or gelatin derived from collagen type III and/or type I monomers (the collagen and gelatin products are collectively hereinafter 25 referred to as "collagen") wherein said monomers are polymerized to form a collagen type III and/or type I composition having sealant properties. Preferably, the collagen is human and is derived using recombinant technology. Collagen type III was selected for its unexpectedly superior hemostatic characteristics, as compared to other collagen types. Collagen type I was selected for its structural characteristics, as well as for the hemostatic 30 properties of certain collagen type I forms (e.g., collagen type I homotrimers). The polymerization reaction may be initiated with an appropriate polymerization initiator such as a chemical oxidant, ultraviolet irradiation, a suitable oxidative enzyme, or atmospheric oxygen. Additionally, cross-linking agents including glutaraldehyde, dye mediated photooxidation , PEG and its derivatives, acyl azide, plyepoxy fixatives, WO 00/09018 PCT/US99/18095 11 5 oxidized starch (periodate) and water soluble carbodiimide ("WSC") may be used in the polymerization process to form a collagen composition having sealant properties. For purposes of optimizing the sealant and adhesive properties of the recombinant collagen product by optimizing the structural stability and the hemostatic characteristics of the product, the product is comprised of a combination of pure recombinant type I and 10 type III collagen The ratio of pure recombinant collagen type III to pure recombinant collagen type I (heterotrimer) is preferably about 30% and greater type III collagen to about 70% or less type I collagen (heterotrimer). More preferably, the ratio of pure recombinant type III collagen to pure recombinant type I collagen (heterotrimer) is about 30% to about 50% type III collagen to about 70% to about 50% type I collagen 15 (heterotrimer). Most preferably, the ratio of pure recombinant type III collagen to pure recombinant type I collagen (heterotrimer) is about 30% to about 40% type III collagen to about 70% to about 60% type I collagen (heterotrimer). With respect to compositions comprised of collagen type I homotrimer, the ratio of pure recombinant type I homotrimer to a combination of recombinant collagen type I 20 heterotrimer and recombinant collagen type III is about 90:10. More preferably, the ratio of pure recombinant type I homotrimer to a combination of recombinant collagen type I heterotrimer and recombinant collagen type III is about 75:25. Most preferably, the ratio of pure recombinant type I homotrimer to a combination of recombinant collagen type I heterotrimer and recombinant collagen type III is about 50:50. 25 It is the object of this invention to provide for a pure recombinant collagen type III tissue sealant, a pure recombinant type I tissue sealant, or a pure recombinant collagen type I and type III tissue sealant, free from other collagen types, having at least one of the following characteristics and capabilities: (i) Hemostasis. The sealant acts as a hemostatic barrier and reduces 30 the risk of serum, lymph, and liquid leakage. As collagen type III possesses inherently hemostatic properties, its use in a hemostatic device provides an improvement over known fibrin sealants. Collagen type I also possesses some hemostatic properties. (ii) Gluing. Due to its adhesive properties, the sealants of the present invention connect tissues by forming a strong joint between them and adapt uneven WO 00/09018 PCT/US99/18095 12 5 wound surfaces. The glueing effect is increased by a combination of agents, such as those described below, and collagen type III and/or collagen type I. (iii) Wound healing. The sealant promotes the growth of fibroblasts which, in combination with efficient hemostasis and adhesion between the wound surfaces provides for an improved healing process. (See also, Ghersi, et al., 1989, Eur. J. 10 Cell Biol. 50:279-284 (comparing characteristics of homotrimer and heterotrimer collagen type I).) The use of the present compositions as anti-adherence/wound healing compositions is expected to result in a normal (regenerative) tissue rather than scar tissue, i.e. optimal wound healing. Furthermore, such compositions also reduce the inflammatory response. 15 Accordingly, it is an object of the present invention to provide polymerized collagen type III and/or type I compositions as safe, effective biological adhesives with appropriate adhesive strength for biomedical applications, particularly those involving soft tissues. More specifically, the present invention is directed to compositions useful in sealing punctures and incisions in large blood vessels and the heart. The polymerized 20 materials may assume a number of sizes and shapes consistent with their intended biomedical applications, which include use in ophthalmology, plastic surgery, orthopedics, and cardiology. The vascular sealant compositions of the present invention, comprising collagen type III and/or I, may be used alone or in combination with a tissue sealant device, including, for example, the devices set forth in U.S. Patent Nos. 5,782,860 25 (issued July 21, 1998), 5,759,194 (issued June 2, 1998), and 5,728,132 (issued March 17, 1998). In another object of the invention, the collagen type III and/or type I composition is further comprised of agents which will confer additional desirable characteristics for a vascular sealant or wound dressing. For example, fibrin, fibrinogen, thrombin, calcium 30 ion, and Factor XIII may be included in the composition to better effect the formation of a three-dimensional network of polymerized collagen. In yet another object of the invention, the recombinant collagen type III composition incorporates a compound having wound healing capabilities. In one embodiment, the compound is connective tissue growth factor and is incorporated in the composition to effect slow-release of the WO 00/09018 PCTIUS99/18095 13 5 compound to the wound. In a second embodiment of the invention, the drug improves vascularization, for example, tumour necrosis factor, as described in U.S. Patent No. 4,808,402 (issued February 28, 1989). BRIEF DESCRIPTION OF THE DRAWINGS 10 Figure 1 shows SDS-PAGE analysis of recombinant type III collagen produced by pichia pastoris. Figure 2 shows data relating to the biocompatibility of recombinant type III collagen and a commercially available collagen hemostat. Figure 3 shows data relating to platelet aggregation experiments of recombinant 1 5 type III and bovine collagen type I. Figure 4 shows data relating to the bleeding time of spleen treated with recombinant collagen type III and bovine collagen type I. Figure 5 shows a SDS-PAGE analysis of bovine collagen I cross-linked with water soluble carbodiimide. 20 Figure 6 shows a SDS-Page analysis of recombinant collagen type III cross-linked with water soluble carbodiimide. DETAILED DESCRIPTION OF THE INVENTION It is understood that the present invention is not limited to the particular 25 methodology, protocols, cell lines, vectors, and reagents, etc., described herein, as these may vary. It is also to be understood that the terminology used herein is used for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention. It must be noted that as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural reference unless the context clearly 30 dictates otherwise. Thus, for example, a reference to "an antibody" is a reference to one or more antibodies and equivalents thereof known to those skilled in the art, and so forth. Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. Preferred methods, devices, and materials are described, although any 35 methods and materials similar or equivalent to those described herein can be used in the WO 00/09018 PCT/US99/18095 14 5 practice or testing of the present invention. All references cited herein are incorporated by reference herein in their entirety. Definitions As employed herein, the term "biologically compatible" refers to recombinant collagen type III and/or type I modified in accordance with the present invention (i.e., a 10 polymerized collagen type III recombinant product) which is incorporated or implanted into or placed adjacent to the biological tissue of a subject and more particularly, does not deteriorate appreciably over time or induce an immune response or deleterious tissue reaction after such incorporation or implantation or placement. As employed herein, the term "pure recombinant collagen type I" refers to 15 collagen type I manufactured by recombinant techniques which is substantially free from other collagen types. Unless otherwise specifically referenced, the term pure recombinant collagen type I includes both collagen type I homotrimer and collagen type I heterotrimer and mixtures thereof. The term includes any other forms of recombinant collagen type I and any modifications made thereto that may be categorized as a subset of collagen, such 20 as gelatins. The term excludes collagen type I isolated from natural sources. As employed herein, the term "pure recombinant collagen type III" refers to human collagen type III manufactured by recombinant techniques which is substantially free from other collagen types. The term includes any other forms of recombinant collagen type III and any modifications made thereto that may be categorized as a subset 25 of collagen, such as gelatins. The term excludes collagen type III isolated from natural sources. As employed herein, the term "substantially free" refers to a recombinant collagen type that is substantially pure of any other collagen type or unmixed with any other collagen type, and is preferably at least 90% free from other collagen types. 30 As employed herein, the term "vascular sealant" refers to any composition useful in closing vascular wounds, including plugs, which possesses hemostatic properties. As employed herein, the term "wound" refers to any opening in the skin, mucosa or epithelial linings, most such openings generally being associated with exposed, raw or abraded tissue. There are no limitations as to the type of wound or other traumata that WO 00/09018 PCT/US99/18095 15 5 can be treated in accordance with this invention, such wounds including, but are not limited to, first, second, and third degree burns (especially second and third degree); surgical incisions, including those of cosmetic surgery; wounds, including lacerations, incisions, and penetrations; and ulcers, including decubital ulcers (bed-sores) and ulcers or wounds associated with diabetic, dental, haemophilic, malignant, and obese patients. 10 Although the primary concern is the healing of major wounds by neovascularization, it is contemplated that the present invention may also be useful for minor wounds, and for cosmetic regeneration of epithelial cells. Preferably, the wounds to be treated are burns and surgical incisions, whether or not associated with viral infections or tumors 15 Preparation of Polymerized Recombinant Collagen Type I and III Production of Collagen Type I and III Monomers. Types of collagen useful in forming the biologically compatible collagen products of the invention with adhesive and hemostatic properties are recombinant collagen type I and type III. Monomeric soluble collagen types I and III is obtained by recombinant processes, including processes 20 involving the production of collagen type III in transgenic animals. Such recombinant processes are set forth, for example, in U.S. Patent No. 5,593,859, which is incorporated herein by reference. Preferably, collagen types I or III will be recombinantly manufactured by culturing a cell which has been transfected with at least one gene encoding the polypeptide comprising collagen type I or III and genes encoding the c and 25 P subunits of the post-translational enzyme prolyl 4-hydroxylase and purifying the resultant collagen monomer therefrom. Preferably, the monomeric soluble collagen type I and III material exhibits a viscous consistency and varying degrees of transparency and clarity. Polymerization Of Collagen Type I and III Monomers. The recombinant 30 collagen type I and III solution may be subsequently subjected to polymerization or cross-linking conditions to produce the polymerized collagen composition of the present invention. Polymerization may be carried out using irradiation, e.g., UV, gamma, or fluorescent light. UV irradiation may be accomplished in the short wave length range using a standard 254 nm source or using UV laser sources. With a standard 254 nm WO 00/09018 PCT/US99/18095 16 5 source, 4-12 watts, polymerization occurs from 10 to 40 minutes, preferably 20 to 30 minutes, at an exposure distance of from 2.5-10 cm, preferably from 2.5 to 5 cm distance. Excess UV exposure will begin to depolymerize the collagen polymers. Polymerization using gamma irradiation can be done using from 0.5 to 2.5 Mrads. Excess gamma exposure will also depolymerize collage polymers. Polymerization in the presence of 10 oxygen can be achieved by adding an initiator to the fluid prior to exposure. Non-limiting examples of initiators include sodium persulfate, sodium thiosulfate, ferrous chloride tetrahydrate, sodium bisulfite, and oxidative enzymes such as peroxidase or catechol oxidase. When initiators are employed, polymerization occurs in 30 seconds to 5 minutes, usually from 1 to 3 minutes. 15 The polymerizing agent is preferably UV irradiation. However, the polymerization or cross-linking of the monomeric substituents can be carried out by any of the methods will known in the art, including simply exposing the material to atmospheric oxygen, although the rate of polymerization is appreciably slower than in the case of UV irradiation or chemical agents. 20 Other agents may also be useful in the polymerization process. For example, to improve the cohesive strength of adhesives formed from the compositions of this invention, difunctional monomeric cross-linking agents may be added to the monomer compositions of this invention to effect polymerization. Such cross-linking agents are known in the art, for example, in U.S. Pat. No. 3,940,362 which is hereby incorporated by 25 reference herein. Additionally, polymerization methods and cross-linking agents such as glutaraldehyde, dye-mediated photooxidation , PEG and its derivatives, acyl azide, plyepoxy fixatives; oxidized starch (periodate) and water soluble carbodiimide ("WSC") well known in the art may be used to produce the polymerized collagen composition of 30 the present invention. (See, e.g., U.S. Patent No. 4,615,794, U.S. Patent No. 5,444,154, U.S. Patent No. 4,500,453, U.S. Patent No. 5,702,818, U.S. Patent No. 5,415,938, U.S. Patent No.5,308,641, U.S. Patent No.5,264,551, U.S. Patent No.5,258,501, U.S. Patent No.5,258,481, U.S. Patent No. 4,427,808, U.S. Patent No. 4,272,610.) WO 00/09018 PCT/US99/18095 17 5 Moreover, the use of polyaldehyde compositions to effectuate polymerization can be also utilized (See, for example, PCT WO 97/29715 and EP 747,066 A2.) Formation of Gelatin. The recombinant collagen protein of the present invention may be further modified and processed into gelatin using procedures known in the art. (See, e.g., Veis, 1965, International Review of Connective Tissue Research, "The 10 Physical Chemistry of Gelatin", Academic Press, New York and London.) For example, a common feature of all standard collagen to gelatin conversion processes is the loss of the secondary structure of the collagen protein, and in the majority of instances, an alteration in either the primary or tertiary structure of the collagen. The collagens of the present invention can be processed using different procedures depending on the type of 15 gelatin desired. In one approach, modifications may occur to unpurified collagen or procollagen present in the cell mass or in the culture medium or any further modifications can be made to the purified collagen as described above. For example, recombinant collagen or procollagen may be modified and processed into recombinant gelatin. Gelatin may be 20 produced directly from the cell mass or the culture medium by taking advantage of gelatin's solubility at elevated temperatures and stability at conditions of low or high pH, low or high salt concentration, and high temperatures. For example, the cell mass or culture medium may further be treated to extract gelatin by denaturing the triple helical structure of collagen using detergents, heat or denaturing agents. (See, e.g., Vies inter alia.) Operations well 25 established for the manufacture of tissue-derived gelatin can be applied to the production of recombinant gelatin. This includes, but is not limited to, treatments with strong alkali or strong acids, heat extraction in aqueous solution, ion exchange chromatography, cross-flow filtration, and heat drying. 30 Collagen Type I and III Compositions The compositions of the present invention are comprised of polymerized type I and III collagen wherein said composition is manufactured by a process comprising the steps of: (1) production of collagen type I and III monomers by the recombinant methods described above; and (2) polymerization of such monomers. In addition, where the final WO 00/09018 PCTIUS99/18095 18 5 composition is a gelatin-based sealant or wound dressing, the process includes a step wherein the collagen is converted into gelatin. For purposes of optimizing the sealant and adhesive properties of the recombinant collagen product by optimizing the structural stability of the product as well as the hemostatic characteristics of the product, the product is comprised preferably of a 10 combination of pure recombinant type I and type III collagen The ratio of pure recombinant collagen type III to pure recombinant type I (heterotrimer) is about 30% and greater type III collagen to about 70% or less type I collagen (heterotrimer). More preferably, the ratio of pure recombinant type III collagen to pure recombinant type I collagen (heterotrimer) is about 30% to about 50% type III collagen to about 70%to about 15 50% type I collagen (heterotrimer). Most preferably, the ratio of pure recombinant type III collagen to pure recombinant type I collagen (heterotrimer) is about 30% to about 40% type III collagen to about 70% to about 60% type I collagen (heterotrimer). The appropriate ranges of concentrations of components in the tissue sealants and adhesives of the present invention can be determined by methods well-known in the art. 20 (See, e.g., Haraski, H. et al. (1999) Volume XXII, Society for Biomaterials, pages 158 through 159; Fasman, G. D., ed. (1989) Practical Handbook of Biochemistry and Molecular Biology, Section 1, pages 126 through 130; U.S. Patent No. 5,834,232 (issued November 10, 1998); Sierra, D. H. et al. (1992) J. Apple. Biomater. 3(2):147-151; Martinowitz, U. and R. Saltz (1996) Curr. Opin. Hematol. 3(5):395-402; and Siriex, D. 25 (1998) Ann. Vasc. Surg. 12(4):311-316.) The actual proportions of the collagen components of the compositions of the present invention will depend on the addition of other agents to the compositions and on the desired use of the compositions. The determination of suitable proportions for particular compositions is within the level of skill in the art, and this invention contemplates the various combinations that can be 30 reached. The compositions of the present invention may be further comprised of other agents useful in gluing or sealing vascular tissues, and more generally, soft tissue. For example, in addition to recombinant collagen type I and/or type III protein, the composition will preferably comprise transglutaminases such as Factor XIII and/or fibrin/ WO 00/09018 PCT/US99/18095 19 5 fibrinogen/fibronectin and/or plasminogen. The suitable concentrations of these components can be selected by methods-well known in the art. For example, fibrinogen can be present in plasma concentrations, such as from about 1.5 to about 4.0 mg/ml, or higher. Fibrinogen can also be present in lower concentrations, for example, to monitor performance. 10 Preferably, the composition will also include clotting enzymes, i.e. thrombin, especially in combination with bivalent calcium, such as calcium chloride. The concentration of calcium chloride can vary, for example, from between 40 mM to 0.2 M, depending on the specific purpose of the tissue adhesive composition. High concentrations of calcium chloride inhibit fibroblast growth and are therefore preferred 15 for anti-adherence applications (fibronectin, which stimulates the growth of fibroblasts, can be absent in such compositions). It may further be valuable to include a fibrinolysis inhibitor, such as a plasmin inhibitor, e.g. aprotinin, aprilotinin, alpha-2-antiplasmin, alpha-2-macroglobulin, alpha-i -antitrypsin, epsilon-aminocaproic or tranexamic acid, or a plasmin activator inhibitor, e.g., PAI-I or PAI-2. 20 While the proportions of the previously known ingredients in the tissue adhesive compositions of the invention may be selected according to methods well-known in the art, the necessary amount of the viscosity-enhancing polymer can readily be determined by a person skilled in the art depending on the particular polymer and the intended use form. Thus, if the concentration and/or molecular weight of the viscosity-enhancing 25 polymer is too low, the viscosity increase will be insufficient, and a too high concentration and/or molecular weight will inhibit the fibrin polymerization and the adhesion to the tissue. By increasing the thrombin concentration, the polymerization of composition of the present invention may be quickened, reducing the time until the glue sets. At low 30 thrombin concentrations, for example, the fibrin of the composition will remain more or less fluid for several minutes after application. A further beneficial effect of increasing the viscosity with a viscosity-enhancing polymer in accordance with the invention is therefore that lower concentrations of thrombin, required in situations where the parts to be sealed require subsequent adaptation even on non-horizontal surfaces, can be used.
WO 00/09018 PCTIUS99/18095 20 5 Likewise, the compositions of the present invention may, rather than including a combination of the agents described herein, include a fusion protein wherein the collagen type I and/or type III and, for example, fibrin, are combined to form one molecule. Such fusion proteins may be manufactured according to recombinant techniques described herein. 10 In a further embodiment of the invention, the composition of the present invention includes agents useful in wound healing, either by inducing or promoting the formation of tissue, or, alternatively, by limiting the formation of fibrotic adhesions. Such agents include antibiotics, or growth factors, such as connective tissue growth factor, described in, for example, U.S. Patent No. 5,408,040 and 5,585,270, incorporated herein by 15 reference. In another embodiment of the invention, the drug improves vascularization, for example, tumour necrosis factor, as described in U.S. Patent No. 4,808,402 (issued February 28, 1989). With respect particularly to the vascular sealant aspect of the present invention, vascular sealant compositions comprising collagen type III and/or I may be used alone or 20 in combination with a tissue sealant device, including, for example, the devices set forth in U.S. Patent Nos. 5,782,860 (issued July 21, 1998), 5,759,194 (issued June 2, 1998) and 5,728,132 (issued March 17, 1998). Fields Of Use The polymerized collagen type III and/or type I products of the present invention 25 may be useful to produce mechanical sealants and adhesive systems. Vascular Adhesive Systems. Fields of application include, but are not limited to, general surgery, dentistry, neurosurgery, plastic surgery, thorax and vascular surgery, abdominal surgery, orthopaedics, accident surgery, gynaecology, urology, and opthalmology. The collagen sealants of the present invention have also been used for 30 local application of drugs, such as antibiotics, growth factors, and cytostatics. Sealant Films and Wound Dressings. In one aspect of the invention, the polymerized collagen products can be made in the form of a sealant film. A collagen based film will be flexible and elastic with the consistency and feel of plastic film, but can exhibit high biological compatibility. Uses of sealant films include, but are not WO 00/09018 PCTIUS99/18095 21 5 limited to, prevention of adhesion formation following tendon surgery (i.e., use as a Wrap around tendons}, use as a synthetic tympanic membrane, and uses as substitute facial tissue and wound dressing components. Additional examples of potential uses of sealant films include, treatment of corneal abrasions, wound closure, coating of catheters and instruments, and use as a material to prevent adhesion formation in tissues and tendons 10 (e.g., peritoneal cavity). Further embodiments of the present invention include sealant and adhesive formulations which can be used in systems specific for delivery of numerous drugs and pharmaceutical compositions, including growth factors, antibiotics, and other biologically beneficial compounds. Such materials can be added to the collagen adhesive or sealant to 15 promote cell migration, cell adhesion, and wound healing. Angioplasty and Angiography. Angiography is a diagnostic procedure whereby dye is injected into an artery, preferably the femoral artery, to detect the presence or absence of coronary disease. Angioplasty, also known as PCTA, is a therapeutic procedure which involves the inflation of a balloon in an artery, such as the coronary 20 artery, for the purpose of relieving arterial blockages. After puncturing the femoral artery, a balloon-catheter is introduced through the femoral artery and navigated through to the coronary artery blocked by atherosclerosis (plaque). Once in position, the balloon is inflated and deflated several times in an effort to open the artery by pushing the fatty material against the vessel walls, allowing for blood to circulate to the affected regions of 25 the heart muscle. Various types of balloon catheters are commonly used in angioplasty and angiography, including over-the-wire catheters which utilize an independent guidewire to the site of the disease; 2) fixed-wire catheters, which combine a balloon catheter with a guidewire into one device; 3) rapid-exchange or single-operator exchange catheters, which are over-the-wire catheters that can be exchanged more conveniently 30 than standard over-the-wire catheters; and 4) perfusion catheters, which allow blood flow during the procedure. A rotational tip catheter removes plaque buildup on arterial walls. These devices utilize a technique called differential cutting. Calcified material is rendered into microscopic particles without damaging the artery due to the elastic nature of the arterial walls.
WO 00/09018 PCT/US99/18095 22 5 Angioplasty is a more invasive and complicated procedure than angiography, requiring the insertion of a larger sheath than that used in angiography. The sheath is used as a vehicle for introducing the catheter into the artery. Additionally, angioplasty also requires the use of blood thinners, such as heparin, to prevent clotting during and after the surgical procedure. The anti-clotting agent prevents the body's natural 10 sealing/clotting mechanism and, thus, sealing punctures requires a significant length of time. According to the present invention, after withdrawing the catheter and other invasive devices from the artery, an adhesive applicator may optionally be inserted into the sheath and placed into a position near to or contacting the puncture in the artery. 15 During the procedure, manual or mechanical pressure is applied to the artery to reduce the flow of blood at the puncture site. If possible, excess blood/fluid is removed from the puncture site. Subsequently, recombinant collagen type III and/or type I monomer of the present invention may be applied to the puncture on the external surface of the artery and/or within the puncture track. The monomer then is polymerized and/or cross-linked 20 by the techniques described herein, for example, UV irradiation, such that polymerization takes place within 0 to 300 seconds, preferably within 0 to 120 seconds, more preferably within 0 to 30 seconds, and most preferably 3 to 10 seconds. By applying the collagen monomer composition on the outside of the artery, the incidence of embolism (blockage of the artery or circulatory system) is virtually eliminated. Alternatively, polymerization 25 may be achieved according to the methods set forth in PCT WO 97/29715 and EP 747,066 A2, incorporated herein by reference. Alternatively, a polymerized collagen type III and/or type I may be used and the polymerization step may be avoided. Because of the bonding strength of the adhesive of the present invention, only small amounts of the adhesive are required to seal a punctured 30 artery. Moreover, because the surgical adhesive according to the present invention can polymerize almost immediately, the adhesive can polymerize on the surface and/or along the puncture track of the artery without penetrating the interior of the artery. Accordingly, large pieces or particles of material will not enter the circulatory system, thereby substantially reducing risk of embolism. Due to the fast and strong bonding of WO 00/09018 PCT/US99/18095 23 5 preferred adhesives of the invention, the patient will need to be immobilized for only-a minimal period of time. Administration Formulations. The tissue treatment composition of the present invention may be presented in the same type of preparations as prior art fibrin sealants. The components 10 may be provided in deep frozen solution form or as lyophilized powders, to be diluted prior to use with appropriate aqueous solutions, e.g. containing aprotinin and calcium ions, respectively. Additionally, the vascular sealants of the present invention may be formulated and shaped in the form of collagen plugs, as described in the art and known to one of ordinary skill in the art. 15 The compositions of the present invention can additionally comprise pharmaceutical agents, such as, for example, an antibiotic or a growth factor, by incorporating the agent into the tissue adhesive so as to be enclosed in the collagen network formed upon application of the tissue adhesive. The agent is thus kept at the site of application while being controllably released from the composition, such as when the 20 composition is used as ocular drops, or a wound healing preparation, etc. As also mentioned above, the pharmaceutically active substance to be released from the present tissue adhesive composition may be the viscosity-enhancing polymer in itself or a substance coupled thereto. A specific example of such a viscosity-enhancing polymer fulfilling the viscosity enhancing requirement as well as having therapeutical and 25 pharmaceutical utility, and in which it may be desired to sustain bioavailability, is hyaluronic acid and salts and derivatives thereof which are easily soluble in water and have an extremely short biological half-life. Thus, in one aspect, compositions of the present invention constitute an advantageous slow-release preparation for proteoglycans such as hyaluronic acid and its salts and derivatives, which considerably increases the 30 bioavailability thereof. Notably, the compositions of the present invention are not restricted to those having adhesive properties. Non-adhesive compositions are also included, especially when these compositions are primarily intended for wound healing. These compositions may in particular include non-adhesive proteins such as albumin and/or growth factors.
WO 00/09018 PCT/US99/18095 24 5 Substantially non-adhesive compositions may also be obtained when the polymer part of the composition inhibits the adhesive properties of the protein part. It should in this context be emphasized that the invention comprises both adhesive and substantially non adhesive compositions, although it has for simplicity reasons often has been referred to as an "adhesive" in this specification. 10 Application Of Compositions. The compositions of the present invention may be applied using a variety of dispensing devices. For example, the surgical adhesive may be applied using the devices set forth in U.S. Patent Nos. 4,900,303 (Lemelson) and 5,372,585 (Tiesenbrun) while monitoring the application process through an optical viewing system. The composition of the present invention may also be applied by the 15 devices set forth in U.S. Patent No. 5,129,882 (Weldon et al.), or the other devices referenced above, or other devices as well known in the art. Compositions according to the present invention may also be applied in conjunction with other sealing means. For example, adhesive compositions may be applied to puncture sites which have been closed using surgical suture or tape, such as in 20 the sealing of a puncture or incision in vascular tissues, including the heart. The adhesive in this instance will provide a complete seal, thereby reducing the risk of body fluid leakage from the organ or vessel, e.g., leakage from artery puncture sites. The surgical adhesive of the present invention may additionally be used in conjunction with other sealing means, such as plugs, and the like. Such techniques are set forth in, for example, 25 U.S. Patent Nos. 4,852,568 (Kensey), 4,890,612 (Kensey), 5,053,046 (Janese), 5,061,274 (Kensey), 5,108,421 (Fowler), 4,832,688 (Sagae et al), 5,192,300 (Fowler), 5,222,974 (Kensey et al.), 5,275,616 (Fowler), 5,282,827 (Kensey et al.), 5,292,332 (Lee), 5,324,306 (Makower et al.), 5,370,660 (Weinstein et al.), and 5,021,059 (Kensey et al.). The subject matter of these patents is incorporated herein by reference. 30 Notably, the compositions of this invention can be used to join together two surfaces by applying the particular composition to at least one of the surfaces. Depending on the particular requirements of the user, the adhesive compositions of this invention can be applied by known means, such as with, for example, a glass stirring rod, sterile brush, or medicine dropper, in many situations, a pressurized aerosol dispensing package is WO 00/09018 PCT/US99/18095 25 5 preferred in which the adhesive composition is in solution with a compatible anhydrous propellant. Aerosol application of the monomers is particularly advantageous for use in hemostasis. Mechanisms for aerosol applications are well known in the art. EXAMPLES 10 The following examples are provided solely to illustrate the claimed invention, and are not intended to limit the scope of the invention. Purification of recombinant collagen type III from yeast expression system. The following protocol was used to purify recombinant human collagen type III 15 ("rhc III" or "Rhc III") from Pichia. I. Resuspend I volume of cell pellets with 7 volume 0.lN HCL; 2. Fill Bead-Beater chamber half full with glass bead just taken from -20C freezer; 3. Fill the chamber with cell suspension; 20 4. Assemble the chamber with Ice-water Jacket; 5. Fill out the jacket with ice water with some sodium chloride; 6. Homogenize the cell pellets for 5 X 1 mins with 5 min. of interval between each 1 min. homogenization; 7. Recover the homogenate by filter through a Buchner Funnel without filter 25 paper; 8. Add pepsin solution to final concentration of 0.2 mg/ml and incubate for 8 hours at 4'C; 9. Centrifuge for 30 min. at 10,000 rpm and collect the supernant (fraction Sl) and pellet (fraction P1); 30 10. Adjust the pH to 7.4 with 1OM NaOH, Incubate overnight at 4 0 C; 11. Add 5M NaCl and HAC to IM NaCl, 0.5M HAC; 12. Incubate at 4'C for 1 hour; .13. Collect the pellets by centrifugation (fraction P2); WO 00/09018 PCTIUS99/18095 26 5 14. Dissolve the pellet in 3 volume 0.1M HCl (depending on the collagen amount, adjust the collagen concentration to about 0.3mg/ml); 15. Add 1.5 volume of 3M urea, 0.3M NaCl, 0.15M Tris, pH 7.4 and adjust the pH to 7.4 with NaOH; 16. Run through DEAE-cellulose column (1.6 X 15 cm) at a flow rate of 0.1 10 0.2ml/min; 17. Collect the flowthrough; 18. Concentrate the collagen by precipitation in 1M NaCl, 0.5M HAC; 19. Redissolve the pellet in 10mM HCI (fraction rhcIII); 20. Dialyze the collagen solution against 10mM HCl if necessary; 15 Characterization of recombinant human collagen type III As defined above, purified rhc III was tested by SDS-PAGE as shown in Figure 1 and amino acid analysis was performed and amino acid composition of purified rhc III is shown as below at TABLE 1. 20 TABLE 1 Amino Acid Composition of rhc III Purified from Pichia Pastoris Amino acid RhC III from Pichia Pastoris Human Collagen III Asp 46 42 Glu 68 71 Hyp 132 125 Ser 36 39 Gly 349 350 His 6 Arg 42 46 Thr 21 13 Ala 92 96 WO 00/09018 PCT/US99/18095 27 Pro 105 107 Val 14 14 Met 7 8 Tyr
-
3 Ile 14 13 Leu 20 22 Hyl 0 5 Lys 44 30 Phe 9 8 Hyp/(Hyp+Pro) 0.557 0.538 5 Biocompatibility and Tissue Response Tests Biocompatibility and tissue response of rhc III and a commeric was tested in a rat subcutaneous model. Rhc III and commercial available Collagen Hemostat were 10 formulated into injectable paste/gel under sterile condition. The rhc III samples was tested to insure the endotoxin level is below the limit. The gel was injected subcutaneously into rats. The preliminary data indicates that Rhc III does not cause any erythema and edema. The gel was dissected in day 2, 7 and 28 after injection and examined histologically with H&E staining. The commercial available collagen hemostat 1 5 has a much stronger tissue response than rhc. A comparison of rhc III with the commercial available collagen hemostat harvested from rats on day 7 is shown in Figure 2. 20 Platelet Aggregation Test Methods. The fibrillogenesis of above described recombinant human collagen type III ("rchIII") was tested according to the following method: First rhc III solution was diluted with 10mM HCl to 1 mg/ml, next, 1/10 volume of 200mM Na2HPO4, pH 11.2 was added. The solution was then mixed well and incubated at room temperature 25 (20-22' C) overnight. Following incubation, the fibril slurry was vortexed before usage.
WO 00/09018 PCTIUS99/18095 28 5 Human Platelet Rich Plasma (PRP) was then prepared from fresh blood of an apparently healthy donor. The PRP was then adjusted to 200K/4 and the collagen fibril slurry was added into the PRP. Following addition of the fibril slurry, the platelet aggregation profile with an aggregometer was detected. The minimal amount of collagen to induce complete platelet aggregation is estimated from the amount of collagen to 10 induce a fall in optical density of at least 30% occurred within 5 minutes. Experimental Results. The platelet aggregation capacity of recombinant human collagen III was compared to the platelet aggregation capacity of bovine skin derived collagen according to the methods set forth above and as more fully described in Balleisen, et al., 1975, Klin. Wschr 53:903-905, incorporated herein by reference. As set 15 forth below in TABLE 2, fibrils generated from recombinant human collagen III has a lower minimal amount in inducing human platelet aggregation than fibrils generated from bovine skin collagen. Recombinant human collagen III also has a shorter onset time to induce platelet aggregation. These results indicate that recombinant human collagen is a more hemostatic than tissue derived collagen. 20 TABLE 2 Collagen samples Minimal Amount for Time to Onset (Second) induction of platelet aggregation (tg) Bovine Skin Collagen 10 48 Recombinant human <5 36 Collagen III from Pichia In a subsequent experiment, platelets were obtained from three healthy donors and were adjusted to 200K/ml in plasma. Collagen fibril slurry was added to the platelet 25 suspension and the aggregation was measured with an Aggregometer. The minimal amount of collagen capable of inducing complete platelet aggregation was determined by stepwise decreasing the amount of collagen added. It was assumed that complete aggregation had occurred when a fall in OD of at least 30% occurred within 5 minutes. It was shown that rhe III has a lower minimal amount to induce complete aggregation of 30 platelet, and indicates that rhc III is more hemostatic than bovine collagen I. This results WO 00/09018 PCT/US99/18095 29 5 depicted in Figure 1 demonstrate that collagen III fibrils are more hemostatic than collagen I fibrils. Hemostatic Effects of Rhc III Sponge Rhe III was prepared as follows: 10 Rhc III was expressed in pichia pastoris. A selected expression clone was cultivated in a bioreactor under defined conditions. Rhc III was purified from the harvested cell pellets by limited pepsin digestion and differential salt precipitation. Purified rhc III was formulated into fibrils at first by neutralization with phosphate buffer and incubation at room temperature overnight. The fibrils were collected by 15 centrifugation and then resuspended in water. After homogenization, the rhc III gel was transferred into a mould and lyophilized into a sponge. This type of sponge has very poor water absorption. Water absorptive capacity is critical for applications of rhe III as a hemostat and vascular sealant. To ensure the compositions of the present invention could satisfy 20 this requirement, a process to formulate water absorptive rhc III sponge was developed. Essentially, a sponge was cross-linked, first with UV irradiation and then with 1% WSC. The residual cross-linking reagent was removed by incubation in PBS and the sponge was washed with water. Cross-linked sponges are lyophilized for animal testing and formulation of tissue sealant. This process not only enhanced the 25 water absorption of rhc III sponge, but also significantly increased the mechanical strength of rhc III sponges. In a control study, bovine collagen I was also formulated into sponges following the same procedures. Acclimatized New Zealand White Rabbits were deeply anesthetized and laparotomies were performed to expose the spleens. Using a scapel blade, uniform 30 incisions about 1.0 cm long and 0.3cm deep were made into the spleens. The incisions were then treated with rhc III sponges or with bovine collagen I sponges. The time interval from the application of the test article or positive control material until the bleeding ceases was recorded. Statistical analysis of the mean time to hemostasis was performed using Anova and Student's two sample T-test. It was observed that the WO 00/09018 PCTIUS99/18095 30 5 bleeding time of spleens treated with rhc III sponges was significantly shorter than that of thos& treated with bovine collagen I. The results are shown in Figure 4. Formulation of RhC III Sealant To prepare a rhc III sealant, a three-step experimental approach was pursued involving: 1) preparing a cross-linked rhc III sponge; 2) coating the sponge with 10 human thrombin; and 3) subsequently coating the sponge with human fibrinogen. A collagen sponge cross-linked with water soluble carbodiimide was rinsed with 100% ethanol and placed in a filtration funnel. Human thrombin suspended in ethanol (25U/ml) was used to coat the sponge by filtration. The amount of thrombin on the sponge was about 1OU/cm 2 . Human fibrinogen dissolved in water at 15 concentration of 4mg/ml was precipitated by mixing with 3 volume of ethanol. The precipitated fibrinogen was filtered through the sponge by vacuum. The amount of fibrinogen on the sponge was about 3mg/ cm' . The coated sponge was lyophilized and used for animal tests. Acclimatized New Zealand White Rabbits were deeply anesthetized and 20 laparotomies were performed to expose kidneys and spleens. Using a scapel blade, incisions about 1.0 cm long and 0.3cm deep were made into the kidneys and/or spleens. The incision was treated with commercial collagen sponge INSTAT or rhc III sealant. The time interval from the application of the test article until the bleeding ceased was recorded. The adhesive capacity of testing articles was estimated by 25 peeling the articles from the test site after hemostasis had been achieved. The results are shown in Table 3 and Table 4. TABLE 3 Hemostatic effect and adhesiveness of rhc III Sealant in Kidney Injury Model 30 Non-treatment Instat TM Rhc III Sealant Animal Numbers 7 4 4 Bleeding Time 384 +/- 169 67 +/-10 <10 seconds* ( Seconds) Adhesiveness** ND + ++ * Bleeding stopped instantly upon application of the test articles ** + weak, ++ medium, +++ strong.
WO 00/09018 PCT/US99/18095 31 5 TABLE 4 Hemostatic Effect and adhesiveness of rhc III Sealant 10 in Spleen Injury Model Non-treatment Instat T M Rhc III Sealant Animal Numbers 7 3 4 Bleeding Time 30.85+/- 9.62 4.27+/-0.47 <10 seconds* (minutes) (minutes) Adhesiveness** ND + ++ * Bleeding stopped instantly upon application of the test articles ** + weak, ++ medium, +++ strong. 15 The results of this study demonstrated that rhc III fibrils are able to induce human platelet aggregation at a lower concentration than bovine collagen I, indicating that an Rhc III sponge stops bleeding in spleen injury models within a shorter time period than bovine collagen sponges. A prototype of rhc III enhanced fibrin sealant showed a superior hemostatic potential in vivo over commercial collagen sponges. 20 Cross-linking Experiments Bovine Collagen Type I A feasibility cross-linking test was conducted with soluble bovine collagen I. Bovine collagen I in 10mM HCl was neutralized with 1/10 volume of 0.2M Na2HPO4, pH 11.2 and incubated overnight at room temperature. The fibrils 25 were washed with water and concentrated to 50 mg/ml by centrifugation. The homogenized gel was transferred to a small cell culture dish and lyophilized to form a sponge. To cross-link the collagen, the bovine collagen sponge was incubated in a solution of water soluble carbodiimide ("WSC") at room temperature overnight, then 30 washed with PBS and water. The cross-linked sponge was then dried and tested for solubility and water absorption, and compared to a commercially available collagen sponge (collagen sponge for Angioseal TM). The data is shown in Table 5. 35 WO 00/09018 PCT/US99/18095 32 5 TABLE 5 Samples Color Solubility in 10mM Water Absorption Wetting HCl (mg/mg) Time % of Control (seconds) Collagen White 1.9% 18.2 5 Sponge for Angioseal TM WSC Treated Vitrogen Sponge 0% White 100% 14.5 >120 0.1% White 11.7% 18.3 >120 0.25% White 2.5% 19.3 100 0.5% White 0% 19.0 40 1% White 0% 18.3 22 2% White 0% 19.8 18 As shown in Table 5 and Figure 5, the cross-linked sponges appeared to be intact after incubation in water for 24 hours at room temperature, while the non-cross-linked 10 collagen sponges were not intact. Using one's fingers,the cross-linked collagen sponges were split to test their mechanical strength. The collagen sponges cross-linked with WSC at a concentration higher than 0.5% demonstrated reasonable mechanical strength. Recombinant Collagen Type III Rhc III collagen fibrils were prepared by neutralizing rhc III in 10mM HCl with 15 1/10 volume of 0.2M Na2HPO4, pH 11.2, and incubated overnight at room temperature. Rhe III fibrils were harvested by centrifugation and washed with water. The fibrils were concentrated to 60 mg/ml and homogenized into a gel. The gel was transferred into a mold and lyophilized into a sponge with a thickness of about 2.5mm. The collagen sponge was cross-linked with 1% WSC in 25% ethanol at room 20 temperature for 16 hours. Ethanol was added to accelerate the wetting of collagen sponges in the reaction solution. The cross-linked sponge was then washed with PBS for 2 hours, and subsequently washed three times with water. After lyophilization, collagen sponges of about 1.4 cm 2 in size were used to test for water reabsorption. In addition, 2 mg of the collagen sponge was used to test for solubility. As shown in Table 6, cross- WO 00/09018 PCTIUS99/18095 33 5 linking of the rhc III sponge improved its water absorptive capacity. The wetting time of the cross-linked rhcIII sponge was reduced to about 10 seconds, and water uptake increased from 76mg to 433mg. The cross-linked rhc III sponge remained intact after incubation in water for 24 hours at room temperature, while the non-cross-linked rhc III did not remain intact. 10 TABLE6 Samples Color Solubility in 10mM Water Wetting HCl Absorption mg Time (% of Control) in 90 seconds (seconds) Collagen Sponge White 1.9% 450 5 for TM Angioseal Rhc III Sponge White 100% 76 > 120 Cross-linked rhc White 5.1% 433 10 III sponge The collagen sponges were also tested with SDS-PAGE. As shown in Figure 6, lane 1 and lane 6 represent rhe III; lane 2 represents the collagen sponge for 1 5 Angioseal
TM
; lane 3 represents cross-linked bovine collagen I sponge; lane 4 represents cross-linked rhc III sponge; and lane 5 represents non-cross-linked rhe III sponge. The collagen sponge Angioseal T M , cross-linked bovine collagen I sponge, and cross-linked rhc III sponge is not soluble in SDS-PAGE buffer. Various modifications and variations of the described methods and systems of the 20 invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific preferred embodiments, it should be understood that the invention as claimed should not be unduly limited to such specific embodiments. Indeed, various modifications of the described modes for carrying out the invention which are obvious to 25 those skilled in molecular biology or related fields are intended to be within the scope of the following claims. All references cited herein are incorporated by reference herein in their entirety.
Claims (53)
1. A sealant composition comprising a polymerized collagen type III wherein 10 said sealant composition is produced by recombinantly manufacturing pure collagen type III monomers in a cell and polymerizing said monomers with an agent.
2. The composition of claim 1, wherein the composition is biologically compatible.
3. The composition of claim 1, wherein the recombinant manufacture of a 15 collagen type III monomer comprises the following steps: (a) culturing a cell which has been transfected with at least one gene encoding a polypeptide comprising collagen type III and at least one gene encoding a polypeptide selected from the group the a or P subunit of prolyl 4-hydroxylase; and (b) purifying said collagen type III. 20
4. The composition of claim 1, wherein the composition is further comprised of one or more agents selected from the group fibrin, fibrinogen, thrombin, Factor XIII, or connective tissue growth factor.
5. The composition of claim 1, wherein the sealant is a vascular sealant.
6. The composition of claim 1, wherein the collagen is gelatin. 25
7. The composition of claim 1, wherein the composition is non-adhesive.
8. The composition of claim 7, wherein the composition is further comprised of albumin.
9. The composition of claim 1, wherein the polymerizing is accomplished by cross-linking. 30
10. A process for making a tissue sealant comprising the steps: (a) manufacturing collagen type III monomers by recombinant means; and (b) polymerizing said collagen type III monomers. WO 00/09018 PCTIUS99/18095 35 5
11. The process of claim 10, wherein the polymerizing is accomplished by cross-liiiking.
12. A sealant composition comprising a polymerized collagen type I wherein said sealant composition is produced by recombinantly manufacturing pure collagen type I monomers in a cell and polymerizing said monomers with an agent. 10
13. The composition of claim 12, wherein the composition is biologically compatible.
14. The composition of claim 12, wherein the recombinant manufacture of a collagen type I monomer comprises the following steps: (a) culturing a cell which has been transfected with at least one gene 15 encoding a polypeptide comprising collagen type I and at least one gene encoding a polypeptide selected from the group the a or P subunit of prolyl 4-hydroxylase; and (b) purifying said collagen type I.
15. The composition of claim 12, wherein the collagen is a gelatin.
16. The composition of claim 12, wherein the composition is further 20 comprised of one or more agents selected from the group fibrin, fibrinogen, thrombin, Factor XIII or connective tissue growth factor.
17. The composition of claim 12, wherein the sealant is a vascular sealant.
18. The composition of claim 12, wherein the collagen type I is a heterotrimer collagen. 25
19. The composition of claim 12, wherein the collagen type I is a homotrimer collagen.
20. The composition of claim 12, wherein the composition is non-adhesive.
21. The composition of claim 20, wherein the composition is further comprised of albumin. 30
22. The composition of claim 12, wherein the polymerizing is accomplished by cross-linking.
23. A process for making a tissue sealant comprising the steps: (a) manufacturing collagen type I monomers by recombinant means; and WO 00/09018 PCT/US99/18095 36 5 (b) polymerizing said collagen type I monomers.
24. The process of claim 23, wherein the polymerizing is accomplished by cross-linking.
25. A tissue sealant composition comprising a polymerized pure collagen type III and a polymerized pure collagen type I. 10
26. The composition of claim 25 wherein the composition is biologically compatible.
27. The composition of claim 25 wherein the ratio of pure recombinant collagen type III to pure recombinant collagen type I is about 30% or greater collagen type III to about 70% or less collagen type I. 15
28. The composition of claim 25 wherein the collagen is a gelatin.
29. The composition of claim 25 wherein the composition is further comprised of one or more agents selected from the group fibrin, fibrinogen, thrombin, Factor XIII or connective tissue growth factor.
30. A wound dressing composition comprising a polymerized collagen type III 20 wherein said composition is produced by recombinantly manufacturing pure collagen type III monomers in a cell and polymerizing said monomers with an agent.
31. The composition of claim 30 wherein the composition is biologically compatible.
32. The composition of claim 30 wherein the recombinant manufacture of a 25 collagen type III monomer comprises the following steps: (a) culturing a cell which has been transfected with at least one gene encoding a polypeptide comprising collagen type III and at least one gene encoding a polypeptide selected from the group the a or P subunit of prolyl 4-hydroxylase; and (b) purifying said collagen type III. 30
33. The composition of claim 30, wherein said collagens are a gelatin.
34. The composition of claim 30, wherein the composition is further comprised of one or more agents selected from the group fibrin, fibrinogen, thrombin, Factor XIII or connective tissue growth factor. WO 00/09018 PCT/US99/18095 37 5
35. The compostion of claim 30, wherein the polymerizing is accomplished by cross-linking.
36. A process for making a wound dressing comprising the steps: (a) manufacturing collagen type III monomers by recombinant means; and 10 (b) polymerizing said collagen type III monomers.
37. The process of claim 36, wherein the polymerizing is accomplished by cross-linking.
38. A wound dressing composition comprising a polymerized collagen type I wherein said composition is produced by recombinantly manufacturing pure collagen 15 type I monomers in a cell and polymerizing said monomers with an agent.
39. The composition of claim 38 wherein the composition is biologically compatible
40. The composition of claim 38 wherein the recombinant manufacture of a collagen type I monomer comprises the following steps: 20 (a) culturing a cell which has been transfected with at least one gene encoding a polypeptide comprising collagen type I and at least one gene encoding a polypeptide selected from the group the a or P subunit of prolyl 4-hydroxylase; and (b) purifying said collagen type I.
41. The composition of claim 38 wherein the said collagen is a gelatin. 25
42. The composition of claim 38 wherein the composition is further comprised of one or more agents selected from the group fibrin, fibrinogen, thrombin, Factor XIII or connective tissue growth factor.
43. The composition of claim 38, wherein the collagen is gelatin.
44. The composition of claim 38, wherein the collagen type I is a heterotrimer 30 collagen.
45. The composition of claim 38, wherein the collagen type I is a homotrimer collagen.
46. The composition of claim 38, wherein the polymerizing is accomplished by cross-linking. WO 00/09018 PCT/US99/18095 38 5
47. A process for making a wound dressing comprising the steps: (a) manufacturing collagen type I monomers by recombinant means; and (b) polymerizing said collagen type I monomers.
48. The process of claim 47, wherein the polymerizing is accomplished by 10 cross-linking.
49. A wound dressing composition comprising a polymerized pure collagen type III and a polymerized pure collagen type I.
50. The composition of claim 49 wherein the composition is biologically compatible. 15
51. The composition of claim 49, wherein the ratio of pure recombinant collagen type III to pure recombinant collagen type I is about 30% or greater collagen type III to about 70% or less collagen type I.
52. The composition of claim 49, wherein said collagens are gelatin.
53. The composition of claim 49, wherein the composition is further 20 comprised of one or more agents selected from the group fibrin, fibrinogen, thrombin, Factor XIII or connective tissue growth factor.
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US9599798P | 1998-08-10 | 1998-08-10 | |
US60095997 | 1998-08-10 | ||
PCT/US1999/018095 WO2000009018A1 (en) | 1998-08-10 | 1999-08-10 | Collagen type i and type iii hemostatic compositions for use as a vascular sealant and wound dressing |
Publications (1)
Publication Number | Publication Date |
---|---|
AU5671899A true AU5671899A (en) | 2000-03-06 |
Family
ID=22254563
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
AU56718/99A Abandoned AU5671899A (en) | 1998-08-10 | 1999-08-10 | Collagen type i and type iii hemostatic compositions for use as a vascular sealant and wound dressing |
Country Status (6)
Country | Link |
---|---|
EP (1) | EP1105051A4 (en) |
JP (1) | JP2002524110A (en) |
AU (1) | AU5671899A (en) |
CA (1) | CA2339575A1 (en) |
MX (1) | MXPA01001510A (en) |
WO (1) | WO2000009018A1 (en) |
Families Citing this family (42)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6599526B2 (en) * | 2000-08-18 | 2003-07-29 | The University Of North Texas Health Science Center At Fort Worth | Pericardial anti-adhesion patch |
JP4199004B2 (en) | 2000-11-07 | 2008-12-17 | クライオライフ、インコーポレイテッド | Foamable foam-like biomaterial and method |
ES2253523T3 (en) * | 2001-01-25 | 2006-06-01 | Nycomed Pharma As | A METHOD FOR PREPARING A COLOGEN SPONGE, A DEVICE FOR REMOVING A PART OF A COLLAGEN FOAM, AND A LONG COLONED SPONGE. |
US7052713B2 (en) | 2001-02-13 | 2006-05-30 | Nycomed Pharma As | Carrier with solid fibrinogen and solid thrombin |
AU2003268127A1 (en) * | 2002-08-20 | 2004-03-11 | Richard L. Grant | Compositions comprising epithelial cells for the treatment and prevention of tissue adhesions |
JPWO2004018615A1 (en) * | 2002-08-23 | 2005-12-08 | 旭化成メディカル株式会社 | Fibrin-containing composition |
JP2006519086A (en) * | 2003-02-28 | 2006-08-24 | ファイブローゲン、インコーポレーテッド | Collagen composition and biomaterial |
AU2004249233A1 (en) * | 2003-06-19 | 2004-12-29 | Vascular Therapies Llc | Medical devices and methods for regulating the tissue response to vascular closure devices |
EP1759718A4 (en) * | 2004-05-21 | 2011-03-02 | Chemo Sero Therapeut Res Inst | Tissue closing preparation |
WO2006107188A1 (en) | 2005-04-06 | 2006-10-12 | Fujifilm Manufacturing Europe B.V. | A non-porous film for culturing cells |
US8227415B2 (en) | 2005-04-06 | 2012-07-24 | Fujifilm Manufacturing Europe B.V. | Non-porous film for culturing cells |
KR20100052499A (en) * | 2007-08-01 | 2010-05-19 | 에디컨인코포레이티드 | Collagen-related peptides and uses thereof |
US8642831B2 (en) | 2008-02-29 | 2014-02-04 | Ferrosan Medical Devices A/S | Device for promotion of hemostasis and/or wound healing |
GB201008404D0 (en) | 2010-05-20 | 2010-07-07 | Fujifilm Mfg Europe Bv | Hemostatic compositions |
WO2012048298A2 (en) | 2010-10-08 | 2012-04-12 | Caridianbct, Inc. | Methods and systems of growing and harvesting cells in a hollow fiber bioreactor system with control conditions |
US11109849B2 (en) | 2012-03-06 | 2021-09-07 | Ferrosan Medical Devices A/S | Pressurized container containing haemostatic paste |
RU2636240C2 (en) | 2012-06-12 | 2017-11-21 | Ферросан Медикал Дивайсиз А/С | Dry haemostatic composition |
US9724078B2 (en) | 2013-06-21 | 2017-08-08 | Ferrosan Medical Devices A/S | Vacuum expanded dry composition and syringe for retaining same |
CN105793411B (en) | 2013-11-16 | 2018-04-17 | 泰尔茂比司特公司 | Cell amplification in bioreactor |
US10111980B2 (en) | 2013-12-11 | 2018-10-30 | Ferrosan Medical Devices A/S | Dry composition comprising an extrusion enhancer |
WO2015148704A1 (en) | 2014-03-25 | 2015-10-01 | Terumo Bct, Inc. | Passive replacement of media |
WO2016049421A1 (en) | 2014-09-26 | 2016-03-31 | Terumo Bct, Inc. | Scheduled feed |
RU2715235C2 (en) | 2014-10-13 | 2020-02-26 | Ферросан Медикал Дивайсиз А/С | Dry composition for use in haemostasis and wound healing |
JP6747650B2 (en) | 2014-12-24 | 2020-08-26 | フェロサン メディカル デバイシーズ エイ/エス | Syringe for holding and mixing the first substance and the second substance |
WO2017004592A1 (en) | 2015-07-02 | 2017-01-05 | Terumo Bct, Inc. | Cell growth with mechanical stimuli |
CN107771093B (en) | 2015-07-03 | 2021-06-15 | 弗罗桑医疗设备公司 | Syringe for mixing two components and for maintaining vacuum under storage conditions |
EP3464565A4 (en) | 2016-05-25 | 2020-01-01 | Terumo BCT, Inc. | Cell expansion |
US11104874B2 (en) | 2016-06-07 | 2021-08-31 | Terumo Bct, Inc. | Coating a bioreactor |
US11685883B2 (en) | 2016-06-07 | 2023-06-27 | Terumo Bct, Inc. | Methods and systems for coating a cell growth surface |
EP3656841A1 (en) | 2017-03-31 | 2020-05-27 | Terumo BCT, Inc. | Cell expansion |
US11624046B2 (en) | 2017-03-31 | 2023-04-11 | Terumo Bct, Inc. | Cell expansion |
US11180541B2 (en) | 2017-09-28 | 2021-11-23 | Geltor, Inc. | Recombinant collagen and elastin molecules and uses thereof |
WO2019167943A1 (en) * | 2018-03-02 | 2019-09-06 | 国立大学法人九州大学 | Tissue-joining member, and use thereof |
MX2020011866A (en) | 2018-05-09 | 2021-01-20 | Ferrosan Medical Devices As | Method for preparing a haemostatic composition. |
CN109675085B (en) * | 2019-02-26 | 2021-09-28 | 百澳瑞派(天津)生物科技有限公司 | Composite collagen dressing for burn wound repair and preparation method thereof |
CN109985271B (en) * | 2019-02-26 | 2021-10-22 | 百澳瑞派(天津)生物科技有限公司 | Composite collagen dressing for healing-difficult wound repair and preparation method thereof |
KR20210151930A (en) | 2019-04-12 | 2021-12-14 | 젤터, 인코포레이티드 | Recombinant elastin and preparation thereof |
CN109821059A (en) * | 2019-04-16 | 2019-05-31 | 大连医科大学附属第一医院 | A kind of preparation method of absorbable fluid gelatin hemostatic material |
US11654057B2 (en) | 2020-04-09 | 2023-05-23 | Bio 54, Llc | Devices for bleeding reduction and methods of making and using the same |
CN114681662A (en) * | 2020-12-30 | 2022-07-01 | 江苏江山聚源生物技术有限公司 | Application of recombinant collagen in preparation of hemostatic material |
WO2023119265A1 (en) | 2021-12-21 | 2023-06-29 | Omrix Biopharmaceuticals Ltd. | Fibrinogen comprising formulation and uses thereof |
US11642324B1 (en) | 2022-03-01 | 2023-05-09 | Bio 54, Llc | Topical tranexamic acid compositions and methods of use thereof |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4852568A (en) * | 1987-02-17 | 1989-08-01 | Kensey Nash Corporation | Method and apparatus for sealing an opening in tissue of a living being |
US5162430A (en) * | 1988-11-21 | 1992-11-10 | Collagen Corporation | Collagen-polymer conjugates |
US5667839A (en) * | 1993-01-28 | 1997-09-16 | Collagen Corporation | Human recombinant collagen in the milk of transgenic animals |
US5759194A (en) * | 1993-09-28 | 1998-06-02 | Hemodynamics, Inc. | Vascular patch applicator |
WO1996021458A1 (en) * | 1995-01-10 | 1996-07-18 | Fibrogen, Inc. | Collagen-based methods and formulations for the treatment of immune system-mediated diseases |
CN1272118A (en) * | 1997-07-28 | 2000-11-01 | 弗勃鲁根股份有限公司 | Collagen type I and type III adhesive compositions |
-
1999
- 1999-08-10 MX MXPA01001510A patent/MXPA01001510A/en unknown
- 1999-08-10 JP JP2000564526A patent/JP2002524110A/en not_active Withdrawn
- 1999-08-10 EP EP99943668A patent/EP1105051A4/en not_active Withdrawn
- 1999-08-10 AU AU56718/99A patent/AU5671899A/en not_active Abandoned
- 1999-08-10 CA CA002339575A patent/CA2339575A1/en not_active Abandoned
- 1999-08-10 WO PCT/US1999/018095 patent/WO2000009018A1/en not_active Application Discontinuation
Also Published As
Publication number | Publication date |
---|---|
WO2000009018A1 (en) | 2000-02-24 |
EP1105051A1 (en) | 2001-06-13 |
MXPA01001510A (en) | 2003-08-20 |
EP1105051A4 (en) | 2003-08-06 |
WO2000009018A9 (en) | 2000-08-03 |
JP2002524110A (en) | 2002-08-06 |
CA2339575A1 (en) | 2000-02-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US20020015724A1 (en) | Collagen type i and type iii hemostatic compositions for use as a vascular sealant and wound dressing | |
AU5671899A (en) | Collagen type i and type iii hemostatic compositions for use as a vascular sealant and wound dressing | |
US20030032143A1 (en) | Collagen type I and type III compositions for use as an adhesive and sealant | |
US20220040371A1 (en) | Photoactivated crosslinking of a protein or peptide | |
JP3735677B2 (en) | Surgical adhesive composition based on non-crosslinked collagen modified by oxidative degradation | |
US5510102A (en) | Plasma and polymer containing surgical hemostatic adhesives | |
US20100297218A1 (en) | Tissue adhesive compositions and methods thereof | |
US10441675B2 (en) | Joining and/or sealing tissues through photo-activated cross-linking of matrix proteins | |
Toriumi et al. | Surgical tissue adhesives in otolaryngology-head and neck surgery | |
AU8662798A (en) | Collagen type i and type iii adhesive compositions | |
Grimaldi et al. | Biotechnological Approaches to Hemostasis and Molecular Mechanisms of Wound Healing | |
MXPA00001013A (en) | Collagen type i and type iii adhesive compositions | |
Lee et al. | Biologic adhesives and hemostatic agents |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
MK5 | Application lapsed section 142(2)(e) - patent request and compl. specification not accepted |