CN1034630C - 促结合组合物的制备方法 - Google Patents
促结合组合物的制备方法 Download PDFInfo
- Publication number
- CN1034630C CN1034630C CN89101581A CN89101581A CN1034630C CN 1034630 C CN1034630 C CN 1034630C CN 89101581 A CN89101581 A CN 89101581A CN 89101581 A CN89101581 A CN 89101581A CN 1034630 C CN1034630 C CN 1034630C
- Authority
- CN
- China
- Prior art keywords
- tooth
- enamel
- protein
- composition
- tissue
- 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.)
- Expired - Lifetime
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 61
- 230000001939 inductive effect Effects 0.000 title abstract description 4
- 210000003298 dental enamel Anatomy 0.000 claims abstract description 50
- 238000000034 method Methods 0.000 claims abstract description 28
- 239000000758 substrate Substances 0.000 claims description 41
- 238000002360 preparation method Methods 0.000 claims description 11
- 235000004252 protein component Nutrition 0.000 claims description 6
- 241000283690 Bos taurus Species 0.000 claims description 4
- 239000003085 diluting agent Substances 0.000 claims description 4
- 241000124008 Mammalia Species 0.000 claims description 3
- 239000011230 binding agent Substances 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 2
- 230000001172 regenerating effect Effects 0.000 claims 1
- 102000004169 proteins and genes Human genes 0.000 abstract description 36
- 108090000623 proteins and genes Proteins 0.000 abstract description 36
- 239000002243 precursor Substances 0.000 abstract description 7
- 239000011159 matrix material Substances 0.000 abstract description 4
- 230000008569 process Effects 0.000 abstract description 3
- 230000008929 regeneration Effects 0.000 abstract 2
- 238000011069 regeneration method Methods 0.000 abstract 2
- 239000000470 constituent Substances 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 description 34
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 33
- 239000000463 material Substances 0.000 description 20
- 238000004108 freeze drying Methods 0.000 description 18
- 230000003239 periodontal effect Effects 0.000 description 18
- 239000000284 extract Substances 0.000 description 17
- 108010022248 tuftelin Proteins 0.000 description 14
- 102000012344 tuftelin Human genes 0.000 description 14
- 210000001519 tissue Anatomy 0.000 description 13
- 201000001245 periodontitis Diseases 0.000 description 12
- 210000000988 bone and bone Anatomy 0.000 description 10
- 238000002474 experimental method Methods 0.000 description 9
- 239000000835 fiber Substances 0.000 description 9
- 210000004195 gingiva Anatomy 0.000 description 9
- 238000005516 engineering process Methods 0.000 description 8
- 230000035876 healing Effects 0.000 description 8
- 210000004379 membrane Anatomy 0.000 description 8
- 238000001816 cooling Methods 0.000 description 7
- 210000000981 epithelium Anatomy 0.000 description 7
- 238000000605 extraction Methods 0.000 description 7
- 239000012528 membrane Substances 0.000 description 7
- 102000008186 Collagen Human genes 0.000 description 6
- 108010035532 Collagen Proteins 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- 229920001436 collagen Polymers 0.000 description 6
- 238000004537 pulping Methods 0.000 description 6
- 150000003839 salts Chemical class 0.000 description 6
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 102100040409 Ameloblastin Human genes 0.000 description 5
- 108010007570 Amelogenin Proteins 0.000 description 5
- 102000007325 Amelogenin Human genes 0.000 description 5
- 101000891247 Homo sapiens Ameloblastin Proteins 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 230000015572 biosynthetic process Effects 0.000 description 5
- 230000006378 damage Effects 0.000 description 5
- 239000006228 supernatant Substances 0.000 description 5
- 210000000689 upper leg Anatomy 0.000 description 5
- 241000282693 Cercopithecidae Species 0.000 description 4
- 108010010803 Gelatin Proteins 0.000 description 4
- 241000700159 Rattus Species 0.000 description 4
- 230000000844 anti-bacterial effect Effects 0.000 description 4
- 238000010612 desalination reaction Methods 0.000 description 4
- 239000008273 gelatin Substances 0.000 description 4
- 229920000159 gelatin Polymers 0.000 description 4
- 235000019322 gelatine Nutrition 0.000 description 4
- 235000011852 gelatine desserts Nutrition 0.000 description 4
- 210000000214 mouth Anatomy 0.000 description 4
- 241001465754 Metazoa Species 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- 108090000190 Thrombin Proteins 0.000 description 3
- 206010052428 Wound Diseases 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 235000014633 carbohydrates Nutrition 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000007710 freezing Methods 0.000 description 3
- 230000008014 freezing Effects 0.000 description 3
- 230000002650 habitual effect Effects 0.000 description 3
- 238000001556 precipitation Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000000523 sample Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 229960004072 thrombin Drugs 0.000 description 3
- 238000002054 transplantation Methods 0.000 description 3
- IXPNQXFRVYWDDI-UHFFFAOYSA-N 1-methyl-2,4-dioxo-1,3-diazinane-5-carboximidamide Chemical compound CN1CC(C(N)=N)C(=O)NC1=O IXPNQXFRVYWDDI-UHFFFAOYSA-N 0.000 description 2
- VLEIUWBSEKKKFX-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;2-[2-[bis(carboxymethyl)amino]ethyl-(carboxymethyl)amino]acetic acid Chemical compound OCC(N)(CO)CO.OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O VLEIUWBSEKKKFX-UHFFFAOYSA-N 0.000 description 2
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 2
- 241000009355 Antron Species 0.000 description 2
- 108010039627 Aprotinin Proteins 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 108010073385 Fibrin Proteins 0.000 description 2
- 102000009123 Fibrin Human genes 0.000 description 2
- 108010080379 Fibrin Tissue Adhesive Proteins 0.000 description 2
- HDSBZMRLPLPFLQ-UHFFFAOYSA-N Propylene glycol alginate Chemical compound OC1C(O)C(OC)OC(C(O)=O)C1OC1C(O)C(O)C(C)C(C(=O)OCC(C)O)O1 HDSBZMRLPLPFLQ-UHFFFAOYSA-N 0.000 description 2
- 229920002472 Starch Polymers 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- 229960004405 aprotinin Drugs 0.000 description 2
- 230000010478 bone regeneration Effects 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000001768 carboxy methyl cellulose Substances 0.000 description 2
- 239000001913 cellulose Substances 0.000 description 2
- 235000010980 cellulose Nutrition 0.000 description 2
- 229920002678 cellulose Polymers 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000000354 decomposition reaction Methods 0.000 description 2
- 210000004268 dentin Anatomy 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 238000002513 implantation Methods 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 208000027866 inflammatory 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
- 230000008774 maternal effect Effects 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 235000010409 propane-1,2-diol alginate Nutrition 0.000 description 2
- 239000000770 propane-1,2-diol alginate Substances 0.000 description 2
- 210000003456 pulmonary alveoli Anatomy 0.000 description 2
- 108010048734 sclerotin Proteins 0.000 description 2
- 235000010413 sodium alginate Nutrition 0.000 description 2
- 239000000661 sodium alginate Substances 0.000 description 2
- 229940005550 sodium alginate Drugs 0.000 description 2
- 239000008107 starch Substances 0.000 description 2
- 235000019698 starch Nutrition 0.000 description 2
- 239000013589 supplement Substances 0.000 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 229920003169 water-soluble polymer Polymers 0.000 description 2
- 230000029663 wound healing Effects 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 108010039209 Blood Coagulation Factors Proteins 0.000 description 1
- 102000015081 Blood Coagulation Factors Human genes 0.000 description 1
- 229920002134 Carboxymethyl cellulose Polymers 0.000 description 1
- 208000032170 Congenital Abnormalities Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 208000006558 Dental Calculus Diseases 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 206010063560 Excessive granulation tissue Diseases 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 108010049003 Fibrinogen Proteins 0.000 description 1
- 102000008946 Fibrinogen Human genes 0.000 description 1
- 229920002907 Guar gum Polymers 0.000 description 1
- 229920002153 Hydroxypropyl cellulose Polymers 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 238000003109 Karl Fischer titration Methods 0.000 description 1
- 229920000161 Locust bean gum Polymers 0.000 description 1
- 229920003091 Methocel™ Polymers 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241001494479 Pecora Species 0.000 description 1
- 208000006735 Periostitis Diseases 0.000 description 1
- 241000337665 Phatnoma Species 0.000 description 1
- 229940124158 Protease/peptidase inhibitor Drugs 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 241001074085 Scophthalmus aquosus Species 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 229920002125 Sokalan® Polymers 0.000 description 1
- 208000002847 Surgical Wound Diseases 0.000 description 1
- 208000008010 Tooth Avulsion Diseases 0.000 description 1
- DPXJVFZANSGRMM-UHFFFAOYSA-N acetic acid;2,3,4,5,6-pentahydroxyhexanal;sodium Chemical compound [Na].CC(O)=O.OCC(O)C(O)C(O)C(O)C=O DPXJVFZANSGRMM-UHFFFAOYSA-N 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- ORILYTVJVMAKLC-UHFFFAOYSA-N adamantane Chemical compound C1C(C2)CC3CC1CC2C3 ORILYTVJVMAKLC-UHFFFAOYSA-N 0.000 description 1
- 229910001573 adamantine Inorganic materials 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000010419 agar Nutrition 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 235000010443 alginic acid Nutrition 0.000 description 1
- 229920000615 alginic acid Polymers 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000002421 anti-septic effect Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000007698 birth defect Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 239000003114 blood coagulation factor Substances 0.000 description 1
- 210000004204 blood vessel Anatomy 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 208000035269 cancer or benign tumor Diseases 0.000 description 1
- 235000010948 carboxy methyl cellulose Nutrition 0.000 description 1
- 239000008112 carboxymethyl-cellulose Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 235000019994 cava Nutrition 0.000 description 1
- 210000001431 cementocyte Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 208000037893 chronic inflammatory disorder Diseases 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 238000011443 conventional therapy Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 210000003074 dental pulp Anatomy 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 229940012952 fibrinogen Drugs 0.000 description 1
- 239000000945 filler Substances 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 210000001126 granulation tissue Anatomy 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000000665 guar gum Substances 0.000 description 1
- 235000010417 guar gum Nutrition 0.000 description 1
- 229960002154 guar gum Drugs 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 239000001863 hydroxypropyl cellulose Substances 0.000 description 1
- 235000010977 hydroxypropyl cellulose Nutrition 0.000 description 1
- 230000007124 immune defense Effects 0.000 description 1
- 230000001771 impaired effect Effects 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 235000010420 locust bean gum Nutrition 0.000 description 1
- 239000000711 locust bean gum Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 230000001089 mineralizing effect Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 210000002200 mouth mucosa Anatomy 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 239000001814 pectin Substances 0.000 description 1
- 235000010987 pectin Nutrition 0.000 description 1
- 229920001277 pectin Polymers 0.000 description 1
- 239000000137 peptide hydrolase inhibitor Substances 0.000 description 1
- 208000028169 periodontal disease Diseases 0.000 description 1
- 210000002379 periodontal ligament Anatomy 0.000 description 1
- 210000003460 periosteum Anatomy 0.000 description 1
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000004584 polyacrylic acid Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 108020001775 protein parts Proteins 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 230000021749 root development Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000003307 slaughter Methods 0.000 description 1
- 235000019812 sodium carboxymethyl cellulose Nutrition 0.000 description 1
- 229920001027 sodium carboxymethylcellulose Polymers 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 229940033618 tisseel Drugs 0.000 description 1
- 230000017423 tissue regeneration Effects 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K6/00—Preparations for dentistry
-
- 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
- A61L24/0005—Ingredients of undetermined constitution or reaction products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/32—Bones; Osteocytes; Osteoblasts; Tendons; Tenocytes; Teeth; Odontoblasts; Cartilage; Chondrocytes; Synovial membrane
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/02—Stomatological preparations, e.g. drugs for caries, aphtae, periodontitis
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
- Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y10S530/00—Chemistry: natural resins or derivatives; peptides or proteins; lignins or reaction products thereof
- Y10S530/827—Proteins from mammals or birds
- Y10S530/84—Bones; tendons; teeth; cartilage
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Rheumatology (AREA)
- Developmental Biology & Embryology (AREA)
- Zoology (AREA)
- Surgery (AREA)
- Virology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Immunology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Organic Chemistry (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Dental Preparations (AREA)
- Materials For Medical Uses (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Adhesives Or Adhesive Processes (AREA)
- Curing Cements, Concrete, And Artificial Stone (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
一种用于促进矿物化组织各部分间结合的组合物，其结合方法是在至少一个部分上再生矿物化组织，该组合物含有作为活性成分的由牙釉质前体，即所谓釉质基质产生的蛋白质组分；一种采用上述组合物通过在至少一个部分上再生矿物化组织促进活体矿物化组织各部分间结合的方法。
Description
本发明涉及一种用于促使矿物化组织各部分之间结合的组合物，其方法为：至少在一个部位，也可能在另一部位上形成新的矿物化组织。本发明还涉及促使此种结合的方法，例如为了牙周炎的治疗。
本发明涉及用于促使矿物化组织各部分(例如：牙齿和骨骼)结合的新生物技术。虽然本发明通常能用于此种结合，但本公开主要解释有关松动牙齿，即所谓的牙周炎的治疗，然而应指出：本发明的主要解释决不能以限定的方式予以理解。在更准确地描述本发明技术之前，为了便于理解，有必要简述一下与牙齿以及相关疾病有关的生物学状态的背景。在正常牙齿状态下，牙齿被固定于颌骨上的特殊腔内，称之为牙槽。在齿根与颌骨之间有一所谓的牙周膜。牙根主要由称为牙质的物质构成。牙质周边包绕了一层薄的牙骨质，大约0.01-1mm厚。首先，在牙骨质上可以见到从牙骨质延伸至牙周膜的胶原纤维，它固定于颌骨上，因此，牙骨质对于牙齿与颌骨的连接极为重要。牙周膜厚约0.2mm，由上述胶原纤维和位于所述纤维之间的血管和神经以及属于这些组织的细胞构成。
颌骨并不一直延伸至牙冠。无颌骨复盖的牙根部位有来自根部牙骨质的纤维延伸至周围的牙龈(齿龈)，这些纤维有助于支撑牙齿，进而稳定牙龈。牙龈以及整个口腔复盖着一层薄上皮。此上皮形成围绕牙齿的圈或套筒，在牙齿与上皮之间，紧贴牙齿处形成一个浅沟。
连接牙齿与颌骨组织的炎性疾病经常发生，世界上大多数人都受到不同程度的侵袭。迄今所采用的治疗方法主要针对阻止疾病的发展过程及尽可能地防止牙齿松动。目前，临床尚无有效的方法，即以此种方式治疗从而能使牙齿重新连接于颌骨。
在此类炎性疾病中的另一个问题是牙齿连接的先天性缺陷。有此缺陷的疾人在早年即出现牙周炎的症状，称为幼年性牙周炎。治疗常常是拔牙，并以昂贵的价格用某种齿桥结构代替。
牙齿表面的细菌引起牙周齿龈的慢性炎症。炎性细胞分泌酶企图杀死细菌，但在这一过程中也破坏了连接牙齿与牙龈及颌骨的胶原纤维，因此，牙根或牙骨表面的细胞常遭破坏，口腔粘膜的上皮沿牙龈逐渐受损，形成称之为牙龈裂隙的病变。在裂隙中，新的细菌有了适宜的生长条件，新的炎性细胞侵入此区，产生牙周膜组织的分解。牙骨质细胞死亡，牙龈区骨骼破坏。这一过程通常发展很慢，但在某些阶段却发展很快，经过一段时间后，患病牙齿将完全失去与颌骨的连接。
目前的治疗主要是去除沉积在牙齿表面的细菌。细菌去除后，牙龈和牙周膜的炎症消失，分解过程就会停止。治疗还针对防止在牙齿表面形成新的细菌沉积物。这样就可阻止牙齿与颌骨连接的破坏，但在治疗过程中并不形成新的牙周膜或新的牙骨质。
关于本发明的研究是基于这一理论：牙骨质的形成始于一层薄的秞质前体物，在牙根发育时，它沿所有牙根表面形成。应该指出：质前体物可以促使牙骨质的形成，这一观点并未得到公认。然而这一理论在共同未决的专利申请中被描述，例如：欧洲专利申请书№878502640。对牙骨质形成机理的进一步研究和试验惊人地揭示出牙釉质前体(所谓的釉质基质)含有一种蛋白成份(作为活性成份)，它可从该釉质基质的有机部分中获得。当人们确认构成该蛋白成份的蛋白生物功能为牙齿釉质形成以及特别是牙釉质的矿物化时，这一发现更加令人惊奇(参考文献：Fischer，L.and Termine，D，Clinical Orthopaedics200，1985，362-85)。釉质基质蛋白由一个高分子量部分和一个低分子量部分构成。已发现，它们的活性构成物不仅由同一低分子量部分构成，而且可转由其活性因子构成。该釉质基质蛋白的低分子量部分由醋酸可提取蛋白构成，该蛋白通常称为釉质生成素(amelogenin)，它的分子量高达约40,000，但主要在约5000至25000范围内。
关于本文所用的名词“釉质前体物”(Precursor toenamel)和“釉质基质”(enamel matrix)参考了两篇文献，其中对上述两个名词的定义给予了充分而清楚的表达，这两篇文献是：
A.R.Ten Cate，Oral Histology，Develo-pment，Strucre and Function，The C.V.Mosby Co.，S：t Louis，USA(1980)pp 182-83。
I.A.Mjor，O.Fejerskov，Human OralEmbryology and Histology，Munksyaard，Copenhagen(1986)pp44-45。
这些文献本文引作对比文献。
关于本发明，已发现：如果牙质被暴露于牙周膜细胞(例如：通过在牙根表面磨一个洞)，在缺乏连接健康牙齿与周围组织的纤维的骨样组织形成时，愈合出现。然而，如果人工产生的洞穴表面复盖有来自牙釉质前体物的活性蛋白部分(下文将牙釉质前体称为釉质基质)时，则可见到粘附组织的正常牙骨质的生成。
釉质基质从哺乳动物中提取更好，如牛或猪种属。在实验中发现，给猴和人类用从其它种属的釉质基质中提取的蛋白部分复盖牙根表面磨的洞穴，可以促使骨质形成。
因此，下述的本发明提供了一个通过至少在一个部位形成新鲜矿物化组织来促使活的矿物化组织之间结合的新技术。这些技术的特征在于，应用来自牙釉质前体即所谓的釉质基质的釉质基质蛋白促使结合。于是，本发明还提供一种用于此用途的组合物，该组合物含有活性组成成份，如釉质基质蛋白或活性因子。
如前所述，本发明特别用于牙科治疗，例如牙周炎，即牙齿松动的治疗，牙齿移植或意外所致牙齿脱落的再置入。然而，本发明尚可促进人工植入物的连接，例如，牙齿植入或人造髋关节。本发明还可用于促使在希望提供腱连接的人工植入物上矿物化组织的形成。
根据本发明技术，应用的蛋白成份最好从某种哺乳动物的釉质基质中获取(如未发育成熟的牙齿)。釉质基质的一个理想来源的屠宰的动物，例如猪和小牛。常应宰杀牙齿尚未发育完全的动物(例如猪应在约半岁时)。优先选择的动物为牛或猪属(即牛或猪)但其它种属亦可，如仍在长牙的羊和鼠类。培养的细胞或用重组DNA技术修饰的细菌(例如，美国专利№4,672,032)也可作为这种蛋白成份的补充来源。
根据本发明，用于治疗的组合物可只由蛋白组份或其活性因子(用水适当混合)构成，但组合物也可包含与载体、稀释剂或粘合剂(例如适合于本技术的改良的纤维素、琼脂、藻酸盐或明胶)结合的蛋白成份。将它应用于牙科时，则采用牙科能接受的载体或稀释剂较为合适。目前最好使用由水溶性聚合物组成的载体。这样的聚合物的例子(而不是加以限制)是羧甲基纤维素钠，微晶纤维素，羟甲基纤维素，羟丙基纤维素，甲基纤维素，高分子量的聚丙烯酸，藻酸钠，藻酸丙二醇酯，黄嗜胶，瓜耳胶，洋槐豆胶，改良的淀粉，明胶，果胶或它们的结合物。这些水溶性聚合物与活性蛋白成份结合后，可任意地被转变成胶体或膜，得到具有优良物理性质容易应用的组合物。为了增加储存的稳定性，该组合物可任意含有稳定剂或防腐剂。
本发明还提供治疗牙周病的方法，包括通过促使牙根牙骨质和颌骨以及连接它们的生理性胶原纤维的形成而重新获得牙齿连接。此方法的特征在于其上皮(如果出现)从齿根被去除，而后，牙根复盖一层从釉质基质中获取的有关蛋白成份。
本发明应用于牙周炎治疗时，最好切开邻近患病牙齿区域的胶原纤维以暴露牙根表面。除去上皮(如果出现的话)，然后在清洁的牙根表面涂一层上述蛋白成份或含有这种蛋白成份作为活性成分的组合物，此后，胶原纤维(牙龈)复位，任意缝合，以便愈合。
如前所述，本发明除能应用于牙周炎的治疗外，尚可用于牙齿的再置入或移植。十岁以下的儿童因意外导致一或数个牙齿脱落，这是相当常见的，主要见于前牙。迅速将脱落牙齿放回原位可获得愈合，并可与颌骨正常连接。但在许多情况下，脱落的牙齿没能迅速放回原位，牙齿在口腔外不适当的介质中存放一段时间(例如空气)，这将破坏牙根表面的牙周膜细胞。牙齿放回口腔时，它将不能获得生理性连接，最终产生松动。迄今，尚无通过连接组织再生而达到永久性牙齿连接的方法。
根据本发明的技术，采用机械或化学方法，以适当的方式，除去脱落牙齿死掉的牙周膜，而后，在裸露的牙齿表面施用含有或由上述蛋白成份组成的组合物，再将牙齿放回牙槽，轻轻固定数周。由于牙根表面施用了蛋白成份或其组合物，牙根表面就会产生新的牙骨质，从而牙齿形成新的连接。
关于牙齿移置，即某一个体的牙齿移植给另一个体，已发现移植牙齿的组织可被受体的免疫防御系统攻击，并在很短时期内被分解。人们试图在免疫相合的个体之间进行移植，但其结果令人失望。长期免疫抑制剂治疗以维持一个或数个牙齿会带来危险的结果，所以目前尚无有效的、长期预后良好的牙齿移植方法。
然而，采用本发明的技术，所设计的方法可以解决这一问题。即从供者体拔出要移植的牙齿，去除牙髓，清洁髓腔，在牙髓腔内用牙根充填剂。用机械或化学方法去除牙周膜，用含有活性蛋白成份的组合物复盖牙根，然后将牙齿放入受者口腔内的新位置，固定牙齿一段时间。由于有蛋白成份的存在就会促使复盖移植的牙齿并起固定作用的内源性矿物化组织再形成。
根据本发明的另一优越方面，含有活性蛋白成份的组合物可用基于纤维蛋白原，XIII因子(它是一种血浆凝血因子)和凝血酶的一种组织粘附物来补充。这种补充组合物由釉质基质、纤维蛋白，XIII因子的预混合物构成，面凝血酶则是在施用组合物于外科部位之前立即加入预混合物中。预混合物中可随意加入抑肽酶，以降低分解速率。此种补充组合物的一个有用的商品为Tisseel，它是一种含两种成份的纤维蛋白密封剂，由奥地利维也纳的IMMUNOAG生产和销售。在应用此组织粘附物时，将蛋白成份、纤维蛋白朊、XIII因子以及任意加入的抑肽酶的预混合物与凝血酶溶液混合，然后速将此组合物施用于外科部位。在治疗牙周炎时，这一技术非常利于手术，并且增加了组合物与牙根的粘附，出血停止，粘膜—骨膜片的复位大大简化，且不用缝合。
可用于本发明所述充组合物的现有优选产品包括纤维素衍生物和藻酸酯(盐)，例如，羧甲基纤维素，藻酸钠或丙二醇藻酸酯。
下文的特殊实施例将对本发明作进一步的说明。根据在猴和人身上进行的牙科实验完成这一举例中所参照的附图代表SDS-page，聚丙烯酰胺凝胶电泳分离。根据分子量从釉质基质得到的两个主要蛋白质部分称作成釉质素(amelogwin)(低分子量)和釉质素(enamelin)(高分子量)。实施例1
制备用于动物实验的提取物
为了满足在动物模型中有活性的釉质基质中的成份，制备了下列实验物质。所有物质均来源于猪釉质基质。
E1，将0.3g基质(蛋白成份27+4％)在3ml0.9％NaCl中成浆，并在冰水冷却下与Polythrone均浆1分钟，时间间隔为10秒。然后将均浆液冷冻干燥，得量：199mg，根据半空细胞法(Lowry)蛋白成份为47％+2％。
E2，将0.3g基质成浆，并按上述方法均浆，然后在水浴上加热4分钟。冷冻干燥，得量为199mg，可溶性蛋白成份为13％+3％。
E3，将0.3g基质在3ml 0.5M乙酸(P.a)中成浆，按前述方法均浆，然后为提取蛋白质将该浆在4℃搅拌24小时。冷却下，以10,000rpm离心10分钟。回收沉淀物，冷冻，将上清液冷冻干燥，得到68mg冷冻干燥物，蛋白成份29％+3％。
E4，在3ml 10％EDTA的0.03M三羟甲基氨基甲烷(Tris)缓冲液(PH7.4)中，将0.3g基质成浆，并在冷却下按前述方法均浆。在冷却下将该均浆搅拌24小时，然后将该均浆液进行冰冻离心进行提取。上清液进行反蒸馏水渗析，并经冷冻干燥。得量：11mg，蛋白质成份33％+2％。
E5，提取经离心(E4)后的沉淀物，具体方法为：在冷却下，用约10倍体积的0.5M乙酸慢搅拌24小时。然后再进行一次离心，将上清液冷冻干燥，得量：109mg，蛋白质成份15％+3％。
E6，将按前述E5离心而得的沉淀冷冻干燥，得量：37mg蛋白质成份：18％+4％。
E7，将0.7g基质在3ml 10％EDTA的0.03M三羟甲基氨基甲烷(Tris)缓冲液(PH7.4)中和0.4mM蛋白酶抑制剂PMSF(苯基甲基磺酰氟)一起成浆，均浆，按前述E4方法提取。离心，并将上清液渗析后进行冷冻干燥，得量：1.1mg，蛋白质成份：41％+2％。
E8，在低温下，用0.5M乙酸提取由按前述E7离心而得的沉淀，时间为48小时，离心，将上清液冷冻干燥，得量：101mg，蛋白质成份17％+1％。
E9，将按前述E8离心而得的沉淀冷冻干燥，得量：40mg，蛋白质成份：30％+3％。
由上述E1-9而得的物质的特征在于：蛋白质成份，蛋白质分子量分布(SDS-page with phast)，蛋白质的等电点和碳水化合物成份。
附图中示出了猪釉质基质提取物的显而易见的分子量组成。经SDS-聚丙烯酰胺胶电泳(SDS-8-25％，Phast，Pharmacia)后，通过激光扫描(LKB UHroscanXL)，蛋白带已移至峰位。根据蛋白(14-90kDa)和多肽(2-14kDa)校准参数(pharmacia)估计分子量。给出提取物E4和E5的线图，提取物E4和E5及其它提取物摘要如下。提取物 蛋 白 成 份 (SDS-Page 分 子 量)No 5/12* 6/14* 10/15* 16 22 24 67kDaE1 x x x x X x XE3 x x x X (x)E4 x x XE5 x x X (x)E7 x x XE8 x x X (x)
*在采用多肽校准kit时所得到的较低分子量和在采用蛋白校准kit时所得到的较高分子量。
X＝大量，x＝小量，(x)＝较小量
本实施例的目的是说明由牙釉质(釉质基质)的母体组织经分步提取而得的E1-E9物质对实验产生的边缘牙周伤口治疗的影响。在猴牙齿的边缘牙周组织造成损伤的方法是：用(牙科)圆头锉除去牙骨质，牙周膜和边缘牙槽骨，形成一个约5mm的颈顶距离(cervico-apical distance)。然后将所述物质施用于实验产生的损伤部位，并使伤口愈合。对照组也同样损伤，但不施用任何物质让其愈合。经过8周的愈和期后，对产生的结果进行组织形态学评价。
以原牙骨质和骨复盖的百分比表示该治愈过程结果(表1)。该治愈过程包括形成新牙骨质粘连层，牙周膜和牙槽骨(新的连接)。
表1物 质 对照 E1 E2 E3 E4 E5 E6 E7 E8 E9治愈(％) 5 93 3 79 4 49 2 0 48 5
因此，在施用物质E1，E3，E5，E8后产生了新的连接。在对照组和用物质E2，E4，E6，E7，E7处理的牙齿则不出现上述效果，损伤依旧，并且只有口腔上皮组织复盖牙齿。这些结果表明：在牙周炎的治疗中，施用釉质基质蛋白的低分子量物质促进组织连接。实施例2
由釉质基质制备脱盐的酸提取物
将类似于前述实施例(E3)制剂的冷冻干燥制剂溶解在0.1M乙酸中，并移(144mg/12ml)至含有SephadexG25细粉和0.1M乙酸的15×540mm柱上。收集第一份富蛋白组份(E10)，冷冻干燥。得到34mg含约72％蛋白质的物质，后者相当于起始原料中的85％蛋白。其余蛋白质存在于盐组份中(63mg冷冻干燥物含有约7％的蛋白质)。
将按这种方法制得的冷冻干燥蛋白质装入10ml的小瓶中(每瓶20mg)，在用于动物实验前进行幅射消毒(35kGy)。
按前述实施例所述方法分析该制剂。结果列于表2。
表2
物 质 对 照 E10
治愈(％) 4 72实施例3
制备纯化蛋白质
将200mg冷冻制剂E3(相同于实施例2中所采用的制剂)溶解在20ml 0.1M乙酸中，并将其加到含有于0.1M乙酸中的Sephadex G-75的25×780mm柱上，温度为4℃。以55ml/h的速度洗脱该柱，收集4ml样品。在280nm(紫外)监测洗脱液，将含有主要洗脱物的样品合并，得到5个组份，后者经电泳分析(SDS-page)，得出如下所示的分子量分布情况。在洗脱色谱中出现5个峰(0-1A)。
洗脱样品
(1)50-60：高分子量蛋白质，“釉质素(ename-lins)”(MW)40,000
(2)62-80：高分子量成釉质素(amelogenin)(MW，约25千道尔顿)。
(3)90-100：中等分子量成釉质素(ameloge-nin)(MW约14千道尔顿)
(4)110-125：低分子量成釉质素(ameloge-nin)(MW约5-10千道尔顿)
(5)130-160：盐
将(2)，(3)，(4)峰的洗脱物冷冻干燥后，分别得到10mg，7mg，12mg高分子量，中等分子量和低分子量的成釉质素蛋白质(蛋白成份＞90％)。
本实验的目的是揭示下列物质对实验性边缘牙周伤口治疗的影响。这些物质包括由母体组织至釉质基质(enamlel matrix)提取的高，中，低分子量成釉质素(Amelogenin)物质。在猴子牙齿的边缘牙周组织造成创伤的具体方法是：用(牙科)圆头锉除去牙骨质，牙周膜和边缘牙槽骨，形成约5mm的颈顶距离。然后将所述物质施用于实验产生的创口并使之治愈。同时对照组也同样造成创面，并在不施用任何物质的条件下使之治愈。经8周的治愈期后，对所得结果进行组织形态学评价(表3)。
表3物 质 对照 成釉质素 高分子量 中等分子量 低分子量治愈(％) 4 78 21 15
因此，施用高分子量成釉质素后出现新的连接，而施用中等分子量成釉质素和低分子量成釉质素则治愈水平较低。这些结果表明：在牙周炎的治疗中，施用由釉质基质(enanel matrix)而得的高分子量组份成釉质素可十分有效地促进组织连接。实施例4
制备釉质基质的酸提取物
将屠杀猪(约6个月，屠杀体重约80kg)的低位颌骨上的软组织剔净，并在屠杀间内冷冻。在出现部分解冻后，从冻结的颌骨中切除适宜的牙胚，并分离釉质基质。
在780ml 0.5M乙酸(PH4.1)中使38g釉质基质成浆，并在冰冷却下均浆化(Homogenisator PolytronPT 10-30)。将该均浆在冷却下搅拌22小时，并于约PH4提取可溶蛋白。将不溶物通过离心除去，并将乙酸溶液冷冻干燥。
得到8.5g含有约20％蛋白质成份的冻干物，其相应蛋白质收率约为50％。
分析冷冻干燥提取物的下列数据：水份，乙酸盐，蛋白质成份(Lowry)，碳水化合物成份(Antron试剂)，氨基酸成份，元素分析，蛋白质分子量分布(SDS-Page)，蛋白质等电点。
在用于临床试验前，将含蛋白质的乙酸溶液经过消毒过滤装入10ml的无菌小瓶中，并在无菌条件下冷冻干燥。
本实施例的目的是揭示E3物质对治愈人类边缘性牙周炎的影响。经Swedish Medical Board和The RegionalEthics Comittee批准后，在对边缘牙周炎患者的常规外科治疗中，将所述组合物用作辅助治疗剂。通过手术除去患者的牙垢和颗粒组织。将E3物质“涂”在裸露的牙根表面，并用粘膜骨膜片复盖。通过定期临床检查，记录袋深，附着水平和龈指数以及口腔内立体X光片检查评价治疗结果。将该结果与早期采用惯用牙周手术和同类患者的相似对照面积所得的定量研究结果比较。
结果表明：E3物质使边缘牙槽骨高度明显增加(4-8mm)，使附着水平明显增加(5-9mm)。这是在惯用牙周外科手术后从未见过的结果。与对惯用牙周外科手术的早期研究相比，这种治疗一般不仅更迅速地改善临床表现，而且还迅速降低边缘袋深。这些结果表明：由釉质基质而得的低分子量蛋白组份具有促进人体新牙周组织附着的能力，这是常规治疗不会出现的结果。
表4
对釉质基质均浆和由釉质均浆而得的蛋白组份的分析分析 釉质基质均化物 酸提取物 脱盐提取物 盐组分元素分析 (E1) (E3) (E10) (实施例2)(％W/W)C 11.3 31 50.4 NDH 1.7 4.4 6.8 NDN 3.9 4.5 15.5 0.7O 9.4 27.5 21 NDS 0.2 0.35 1.3 0.1Cl 5.5 0.6 ＜0.1 1.3P 8.8 3.5 0.1 4.3Ca 11.9 1.2 ＜0.1 31.6K 0.2 0.4 ＜0.1 ND
表4(续)
对釉质基质均浆和由釉质均浆而得的蛋白组份的分析分 析 釉质基质均化物 酸提取物 脱盐提取物 盐组分
(E1) (E3) (E10) (实施例2)蛋白质组份(％W/W)Lowry分析 23 20 72 ＜4氨基酸分析 ＜23 ＜26 ＜90 ND碳水化合物组份(％W/W)Antron试剂 ND 0.4 1.3 ND水份(％W/W)Karl Fischer滴 定 ND 2 6 ND乙酸盐组份(％W/W)GC-方法 ND 38 4 ND
表4(续)
对釉质基质均浆和由釉质均浆而得的蛋白组份的分析分 析 釉质基质 酸提取物 脱盐提取物 盐提取物
均浆(E1) (E3) (E10) (实施例2)氨基酸分析(残余物％W/W总残留物)
Pro ND 18.0 19.0 ND
Glu ″ 17.8 19.0 ″
Leu ″ 9.0 9.1 ″
His ″ 8.7 9.2 ″
Ser ″ 4.8 4.6 ″
Gly ″ 3.2 2.8 ″
Tyr ″ 6.4 5.3 ″
Thr ″ 3.6 3.4 ″
Val ″ 3.6 3.5 ″
Met ″ 4.3 5.3 ″
Ile ″ 3.5 3.6 ″
Asp ″ 3.6 3.0 ″
Phe ″ 3.8 3.7 ″
Ala ″ 1.6 1.5 ″
Lys ″ 2.5 1.8 ″
Arg ″ 3.0 2.5 ″
Trp ″ 2.6 2.5 ″
Cys ″ 0 0 ″改善骨骼治疗
根据成年大鼠下颌和股骨的实验洞穴测定“釉质基质低分子量蛋白质”的骨骼治疗作用。在皮肤和咬肌作垂直外科切口使颌骨的角形部分暴露。在恒定的生理盐水流下，在下颌骨上钻一个小孔(直径为2mm)。以与颌骨同样的方法，在股骨侧远部分的致密骨钻几个同样大小的洞。在右颌骨和右股骨施用“釉质基质低分子量蛋白质”，与此同时将左颌骨和左股骨用作对照洞穴。
在这些洞穴中施用的“釉质基质低分子量蛋白质”既可采用冷冻干燥海棉样材料的形式，也可采用小明胶圆柱体的形式。用没有“釉质基质低分子量蛋白质”的明胶圆柱体填充这些大鼠左侧颌骨和股骨上的对照洞穴。在大鼠左侧颌骨和股骨对照洞穴不施用任何干燥，冷冻干燥“釉质基质低分子量蛋白质”。
施用上述物质1-5周后将大鼠处死。取出实验和对照的观察部位并作显微镜检查。
在钻空中施用釉质低分子量蛋白仅1周后，这个空洞就会完全被骨质充满，另外，已形成明显的骨膜对合。在未用本发明方法处理的对照组，也长出新骨，但明显要少许多，且并不能使钻空治愈。
Claims (3)
1.一种用于诱发哺乳动物各部分矿物化组织间结合的组合物的制备方法，所述结合方法是在至少一个部分上再生矿物化组织，该制备方法的特征在于从哺乳动物颌骨分离牙胚，从釉质组织游离该牙胚，从所游离牙胚中回收釉质基质，使釉质基质均浆化并从该均浆化的釉质基质中分离蛋白质组份，并可任意地将该组份与适用于该目的的载体，稀释剂或粘合剂混合。
2.根据于权利要求1所述的方法，其中从牛或猪体内进行所述分离。
3.根据于权利要求1或2所述的方法，其中所述载体，稀释剂或粘合剂是牙科或生物上可以接受的。
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SE8800980-6 | 1988-03-17 | ||
| SE8800980A SE461313B (sv) | 1988-03-17 | 1988-03-17 | Bindningsinducerande komposition |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1035773A CN1035773A (zh) | 1989-09-27 |
| CN1034630C true CN1034630C (zh) | 1997-04-23 |
Family
ID=20371730
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN89101581A Expired - Lifetime CN1034630C (zh) | 1988-03-17 | 1989-03-17 | 促结合组合物的制备方法 |
Country Status (29)
| Country | Link |
|---|---|
| US (2) | US5071958A (zh) |
| EP (1) | EP0337967B1 (zh) |
| JP (1) | JP2795667B2 (zh) |
| KR (1) | KR0175138B1 (zh) |
| CN (1) | CN1034630C (zh) |
| AT (1) | ATE89156T1 (zh) |
| AU (1) | AU613161B2 (zh) |
| CA (1) | CA1339338C (zh) |
| CZ (1) | CZ284330B6 (zh) |
| DD (2) | DD281116A5 (zh) |
| DE (1) | DE68906455T2 (zh) |
| DK (1) | DK175731B1 (zh) |
| ES (1) | ES2055159T3 (zh) |
| FI (1) | FI90011C (zh) |
| HU (2) | HU206611B (zh) |
| IE (1) | IE63098B1 (zh) |
| IL (1) | IL89460A (zh) |
| MX (1) | MX170499B (zh) |
| MY (1) | MY104992A (zh) |
| NO (1) | NO178284C (zh) |
| NZ (1) | NZ228309A (zh) |
| PH (1) | PH25000A (zh) |
| PL (1) | PL160475B1 (zh) |
| RU (1) | RU2008915C1 (zh) |
| SE (1) | SE461313B (zh) |
| SK (1) | SK163789A3 (zh) |
| WO (1) | WO1989008441A1 (zh) |
| YU (1) | YU55389A (zh) |
| ZA (1) | ZA891818B (zh) |
Families Citing this family (26)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5540921A (en) * | 1993-03-10 | 1996-07-30 | Kose Corporation | Solid o/w-type cosmetic composition |
| WO1997002730A2 (en) * | 1995-07-13 | 1997-01-30 | Biora Ab | Enamel matrix related polypeptide |
| US6300062B1 (en) | 1995-07-13 | 2001-10-09 | Biora Ab | Enamel matrix related polypeptide |
| US5762502A (en) * | 1996-07-11 | 1998-06-09 | Bahn; Arthur N. | Process for adhering composites to human teeth |
| DE69910622T2 (de) * | 1998-02-27 | 2004-06-17 | Biora Bioex Ab | Matrixprotein enthaltende Zusammensetzungen für entzündliche und infektiöse Zustände |
| BRPI9908182C1 (pt) | 1998-02-27 | 2021-05-25 | Biora Ab | composições de proteína de matriz para cura de feridas |
| WO2000006734A1 (en) * | 1998-07-29 | 2000-02-10 | Northwestern University | Chondrogenic and osteogenic inducing molecule |
| US6677306B1 (en) | 1998-07-29 | 2004-01-13 | Northwestern University | Chondrogenic and osteogenic inducing molecule |
| ATE258442T1 (de) * | 1999-03-10 | 2004-02-15 | Biora Bioex Ab | Matrixprotein zusammensetzungen für pfropfung in nicht mineralisierten geweben |
| US20030096740A1 (en) | 1999-03-10 | 2003-05-22 | Lars Hammarstrom | Matrix protein compositions for induction of apoptosis |
| DE60140191D1 (de) * | 2000-06-20 | 2009-11-26 | Straumann Inst Ag | Zusammensetzungen mit matrixproteinen zur regeneration von dentin |
| US6899915B2 (en) * | 2000-11-29 | 2005-05-31 | President And Fellows Of Harvard College | Methods and compositions for culturing a biological tooth |
| KR100450146B1 (ko) * | 2002-02-19 | 2004-09-30 | 정필훈 | 발치된 치아로부터 치아 단백질의 추출방법과 그의이용방법 |
| JP4643932B2 (ja) * | 2004-06-28 | 2011-03-02 | 花王株式会社 | アメロゲニン融合タンパク質 |
| US8071131B2 (en) | 2004-07-15 | 2011-12-06 | Ivoclar Vivadent, Inc. | Mineralizing composite materials for restoring teeth |
| KR101206345B1 (ko) | 2004-12-10 | 2012-12-06 | 스트라우만 홀딩 에이쥐 | 신규한 단백질 배합물 |
| SE0403014D0 (sv) | 2004-12-10 | 2004-12-10 | Straumann Holding Ag | New protein formulation |
| US20060257492A1 (en) * | 2005-05-13 | 2006-11-16 | Depuy Products, Inc. | Suspension of calcium phosphate particulates for local delivery of therapeutic agents |
| WO2011051457A2 (en) | 2009-11-02 | 2011-05-05 | Straumann Holding Ag | Purified emd protein composition |
| WO2011073447A1 (en) * | 2009-12-18 | 2011-06-23 | Straumann Holding Ag | Emd c-depleted |
| GB0922438D0 (en) | 2009-12-22 | 2010-02-03 | Ucl Business Plc | Agents having tissue generative activity |
| EP2622063A4 (en) | 2010-10-01 | 2014-03-26 | Univ Columbia | PRODUCTION OF DENTINE, CEMENT AND ENAMEL BY CELLS |
| CN103153334B (zh) | 2010-10-15 | 2016-11-16 | 斯特劳曼控股公司 | 包含pga的新的emd制剂 |
| US9957314B2 (en) | 2011-05-09 | 2018-05-01 | Hadasit Medical Research Services And Development Ltd. | Regeneration and repair of mesenchymal tissue using amelogenin |
| WO2012153333A1 (en) | 2011-05-09 | 2012-11-15 | Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. | Regeneration and repair of mesenchymal tissue using amelogenin |
| US11457996B2 (en) | 2019-09-12 | 2022-10-04 | King Abdulaziz University | Dental pulp capping composition and method of preserving and regenerating capped pulp and dentin bridge |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0128706A2 (en) * | 1983-06-10 | 1984-12-19 | University Patents, Inc. | Body implants of extracellular matrix and means and methods of making and using such implants |
| WO1985002199A1 (en) * | 1983-11-09 | 1985-05-23 | University Of Southern California | Dental enamel production |
| EP0178234A1 (fr) * | 1984-08-31 | 1986-04-16 | Jean-Jacques Russier | Joint physiologique étanche pour implants endo-extracorporels |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| DE2220063A1 (de) * | 1972-04-24 | 1973-11-08 | Ministerul En Electrice | Implantat, seine herstellung und konditionierung |
| US3913229A (en) * | 1974-02-25 | 1975-10-21 | Miter Inc | Dental treatments |
| US4172128A (en) * | 1975-03-26 | 1979-10-23 | Erhard Thiele | Process of degrading and regenerating bone and tooth material and products |
| JPS6045602B2 (ja) * | 1978-09-28 | 1985-10-11 | 正隆 片桐 | 生物性移植体とその製作法 |
| DE3127270A1 (de) * | 1981-07-10 | 1983-03-24 | Link, Gottlieb, 4950 Minden | Knochenersatz-implantat als grundlage fuer die einpflanzung eines kunstzahns |
| JPS595109A (ja) * | 1982-06-30 | 1984-01-12 | Lion Corp | 口腔用組成物 |
| US4702734A (en) * | 1986-05-08 | 1987-10-27 | Meloy Laboratories, Inc. | Method of promoting periodontal regeneration and fibroblast bonding |
| SE454480B (sv) * | 1986-09-25 | 1988-05-09 | Lars Hammarstrom | Bindningsinducerande komposition |
-
1988
- 1988-03-17 SE SE8800980A patent/SE461313B/sv not_active IP Right Cessation
- 1988-03-25 HU HU883310A patent/HU206611B/hu not_active IP Right Cessation
- 1988-03-25 DD DD88314040A patent/DD281116A5/de not_active IP Right Cessation
-
1989
- 1989-03-02 IL IL89460A patent/IL89460A/xx unknown
- 1989-03-09 PH PH38310A patent/PH25000A/en unknown
- 1989-03-09 ZA ZA891818A patent/ZA891818B/xx unknown
- 1989-03-13 NZ NZ228309A patent/NZ228309A/en unknown
- 1989-03-15 PL PL1989278257A patent/PL160475B1/pl unknown
- 1989-03-15 DD DD89326622A patent/DD283563A5/de unknown
- 1989-03-16 HU HU892558A patent/HU203964B/hu unknown
- 1989-03-16 IE IE84689A patent/IE63098B1/en not_active IP Right Cessation
- 1989-03-16 JP JP1062348A patent/JP2795667B2/ja not_active Expired - Lifetime
- 1989-03-16 CZ CS891637A patent/CZ284330B6/cs not_active IP Right Cessation
- 1989-03-16 EP EP89850090A patent/EP0337967B1/en not_active Expired - Lifetime
- 1989-03-16 WO PCT/SE1989/000139 patent/WO1989008441A1/en active IP Right Grant
- 1989-03-16 SK SK1637-89A patent/SK163789A3/sk unknown
- 1989-03-16 AT AT89850090T patent/ATE89156T1/de not_active IP Right Cessation
- 1989-03-16 DE DE8989850090T patent/DE68906455T2/de not_active Expired - Lifetime
- 1989-03-16 DK DK198901280A patent/DK175731B1/da not_active IP Right Cessation
- 1989-03-16 CA CA000593963A patent/CA1339338C/en not_active Expired - Lifetime
- 1989-03-16 AU AU31405/89A patent/AU613161B2/en not_active Expired
- 1989-03-16 ES ES89850090T patent/ES2055159T3/es not_active Expired - Lifetime
- 1989-03-17 KR KR1019890702137A patent/KR0175138B1/ko not_active IP Right Cessation
- 1989-03-17 YU YU00553/89A patent/YU55389A/xx unknown
- 1989-03-17 MY MYPI89000336A patent/MY104992A/en unknown
- 1989-03-17 MX MX015325A patent/MX170499B/es unknown
- 1989-03-17 CN CN89101581A patent/CN1034630C/zh not_active Expired - Lifetime
-
1990
- 1990-09-12 US US07/581,031 patent/US5071958A/en not_active Expired - Lifetime
- 1990-09-13 FI FI904521A patent/FI90011C/fi not_active IP Right Cessation
- 1990-09-14 NO NO904023A patent/NO178284C/no not_active IP Right Cessation
- 1990-09-17 RU SU904831270A patent/RU2008915C1/ru active
-
1991
- 1991-04-22 US US07/689,122 patent/US5098891A/en not_active Expired - Lifetime
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0128706A2 (en) * | 1983-06-10 | 1984-12-19 | University Patents, Inc. | Body implants of extracellular matrix and means and methods of making and using such implants |
| WO1985002199A1 (en) * | 1983-11-09 | 1985-05-23 | University Of Southern California | Dental enamel production |
| EP0178234A1 (fr) * | 1984-08-31 | 1986-04-16 | Jean-Jacques Russier | Joint physiologique étanche pour implants endo-extracorporels |
Also Published As
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN1034630C (zh) | 促结合组合物的制备方法 | |
| JP3471967B2 (ja) | ハイブリッド型人工歯根 | |
| AU593094B2 (en) | Marrow/collagen/mineral matrix for bone defect repair | |
| CA1293927C (en) | Bonding-inducing composition containing enamel matrix | |
| JPH0717876A (ja) | 歯周疾患治療剤 | |
| JP4836318B2 (ja) | 歯根膜形成能を有する人工歯根 | |
| JP3668048B2 (ja) | 歯周病治療材 | |
| FI91216B (fi) | Menetelmä sitoutumista indusoivan koostumuksen valmistamiseksi | |
| JPH0326616B2 (zh) | ||
| CZ193588A3 (cs) | Prostředek pro regeneraci mineralizované tkáně a způsob jeho výroby | |
| PL154615B1 (pl) | Sposób otrzymywania matrycy szkliwa |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C53 | Correction of patent of invention or patent application | ||
| CB02 | Change of applicant information |
Address after: The Netherlands Antilles Curacao Diller Tekade 58a Iran Applicant after: Biora AB Address before: Swedish tile is 47 Applicant before: Biora AB |
|
| COR | Change of bibliographic data |
Free format text: CORRECT: APPLICANT; FROM: BIORA AB TO: BIOLOGICAL INVESTMENT CO. LTD. |
|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C53 | Correction of patent of invention or patent application | ||
| COR | Change of bibliographic data |
Free format text: CORRECT: PATENTEE; FROM: BIOLOGICAL INVESTMENT CO. LTD. TO: BIOLOGY INVESTMENT CO.,LTD. |
|
| CP03 | Change of name, title or address |
Address after: Holland's-Hertogenbosch denbosch Patentee after: Bio investment company Address before: Holland, Netherlands Antilles Curacao Patentee before: Biora AB |
|
| C15 | Extension of patent right duration from 15 to 20 years for appl. with date before 31.12.1992 and still valid on 11.12.2001 (patent law change 1993) | ||
| OR01 | Other related matters | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20090109 Address after: Basel Patentee after: Institut Straumann AG Address before: Holland's-Hertogenbosch denbosch Patentee before: Bio investment company |
|
| ASS | Succession or assignment of patent right |
Owner name: INSTITUT STRAUMANN AG Free format text: FORMER OWNER: BIOLOGICAL INVESTMENT CO.,LTD. Effective date: 20090109 |
|
| C17 | Cessation of patent right | ||
| CX01 | Expiry of patent term |