CN116212111B - 软骨修复用明胶聚己内酯蛋白肽复合气凝胶及制备方法 - Google Patents
软骨修复用明胶聚己内酯蛋白肽复合气凝胶及制备方法 Download PDFInfo
- Publication number
- CN116212111B CN116212111B CN202310254070.9A CN202310254070A CN116212111B CN 116212111 B CN116212111 B CN 116212111B CN 202310254070 A CN202310254070 A CN 202310254070A CN 116212111 B CN116212111 B CN 116212111B
- Authority
- CN
- China
- Prior art keywords
- polycaprolactone
- gelatin
- filtrate
- solution
- hours
- 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.)
- Active
Links
- 229920001610 polycaprolactone Polymers 0.000 title claims abstract description 97
- 239000004632 polycaprolactone Substances 0.000 title claims abstract description 91
- 108010010803 Gelatin Proteins 0.000 title claims abstract description 59
- 229920000159 gelatin Polymers 0.000 title claims abstract description 59
- 239000008273 gelatin Substances 0.000 title claims abstract description 59
- 235000019322 gelatine Nutrition 0.000 title claims abstract description 59
- 235000011852 gelatine desserts Nutrition 0.000 title claims abstract description 59
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 54
- 210000000845 cartilage Anatomy 0.000 title claims abstract description 51
- 230000008439 repair process Effects 0.000 title claims abstract description 48
- 239000002131 composite material Substances 0.000 title claims abstract description 35
- 239000004964 aerogel Substances 0.000 title claims abstract description 34
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 32
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 32
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 102000008186 Collagen Human genes 0.000 claims abstract description 26
- 108010035532 Collagen Proteins 0.000 claims abstract description 26
- 229920001436 collagen Polymers 0.000 claims abstract description 26
- 238000010041 electrostatic spinning Methods 0.000 claims abstract description 22
- 210000001519 tissue Anatomy 0.000 claims abstract description 8
- 239000000243 solution Substances 0.000 claims description 71
- 239000000706 filtrate Substances 0.000 claims description 61
- 229920001661 Chitosan Polymers 0.000 claims description 60
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 47
- 238000003756 stirring Methods 0.000 claims description 41
- 238000001914 filtration Methods 0.000 claims description 39
- 238000002156 mixing Methods 0.000 claims description 38
- 239000000835 fiber Substances 0.000 claims description 35
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 30
- 238000001035 drying Methods 0.000 claims description 30
- 239000002904 solvent Substances 0.000 claims description 29
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 28
- 210000000988 bone and bone Anatomy 0.000 claims description 26
- 241000283690 Bos taurus Species 0.000 claims description 25
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 24
- 238000001704 evaporation Methods 0.000 claims description 24
- DKGAVHZHDRPRBM-UHFFFAOYSA-N Tert-Butanol Chemical compound CC(C)(C)O DKGAVHZHDRPRBM-UHFFFAOYSA-N 0.000 claims description 23
- 239000007864 aqueous solution Substances 0.000 claims description 23
- WFDIJRYMOXRFFG-UHFFFAOYSA-N Acetic anhydride Chemical compound CC(=O)OC(C)=O WFDIJRYMOXRFFG-UHFFFAOYSA-N 0.000 claims description 21
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 21
- 239000002244 precipitate Substances 0.000 claims description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- 239000012528 membrane Substances 0.000 claims description 17
- 238000000034 method Methods 0.000 claims description 17
- 238000006243 chemical reaction Methods 0.000 claims description 16
- ZMANZCXQSJIPKH-UHFFFAOYSA-N Triethylamine Chemical compound CCN(CC)CC ZMANZCXQSJIPKH-UHFFFAOYSA-N 0.000 claims description 15
- 238000001816 cooling Methods 0.000 claims description 15
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 14
- WGQKYBSKWIADBV-UHFFFAOYSA-N benzylamine Chemical compound NCC1=CC=CC=C1 WGQKYBSKWIADBV-UHFFFAOYSA-N 0.000 claims description 14
- 238000004108 freeze drying Methods 0.000 claims description 14
- 239000006228 supernatant Substances 0.000 claims description 14
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 14
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 claims description 12
- 239000012044 organic layer Substances 0.000 claims description 12
- 238000000108 ultra-filtration Methods 0.000 claims description 12
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 11
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 10
- 108090000790 Enzymes Proteins 0.000 claims description 10
- 102000004190 Enzymes Human genes 0.000 claims description 10
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 10
- 102000004142 Trypsin Human genes 0.000 claims description 10
- 108090000631 Trypsin Proteins 0.000 claims description 10
- 229940088598 enzyme Drugs 0.000 claims description 10
- 239000000843 powder Substances 0.000 claims description 10
- 239000012588 trypsin Substances 0.000 claims description 10
- BYEAHWXPCBROCE-UHFFFAOYSA-N 1,1,1,3,3,3-hexafluoropropan-2-ol Chemical compound FC(F)(F)C(O)C(F)(F)F BYEAHWXPCBROCE-UHFFFAOYSA-N 0.000 claims description 9
- 102100026189 Beta-galactosidase Human genes 0.000 claims description 9
- 108010059881 Lactase Proteins 0.000 claims description 9
- 108010005774 beta-Galactosidase Proteins 0.000 claims description 9
- 229940116108 lactase Drugs 0.000 claims description 9
- 125000000218 acetic acid group Chemical group C(C)(=O)* 0.000 claims description 8
- 238000004440 column chromatography Methods 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- 239000012299 nitrogen atmosphere Substances 0.000 claims description 8
- 239000012074 organic phase Substances 0.000 claims description 8
- 239000003480 eluent Substances 0.000 claims description 7
- 238000010438 heat treatment Methods 0.000 claims description 7
- 239000002808 molecular sieve Substances 0.000 claims description 7
- 239000012071 phase Substances 0.000 claims description 7
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 claims description 7
- PAPBSGBWRJIAAV-UHFFFAOYSA-N ε-Caprolactone Chemical compound O=C1CCCCCO1 PAPBSGBWRJIAAV-UHFFFAOYSA-N 0.000 claims description 7
- 230000000415 inactivating effect Effects 0.000 claims description 6
- 230000001678 irradiating effect Effects 0.000 claims description 6
- VHRYZQNGTZXDNX-UHFFFAOYSA-N methacryloyl chloride Chemical compound CC(=C)C(Cl)=O VHRYZQNGTZXDNX-UHFFFAOYSA-N 0.000 claims description 6
- YOCIJWAHRAJQFT-UHFFFAOYSA-N 2-bromo-2-methylpropanoyl bromide Chemical compound CC(C)(Br)C(Br)=O YOCIJWAHRAJQFT-UHFFFAOYSA-N 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 229910018072 Al 2 O 3 Inorganic materials 0.000 claims description 3
- 229910021591 Copper(I) chloride Inorganic materials 0.000 claims description 3
- 239000012300 argon atmosphere Substances 0.000 claims description 3
- OXBLHERUFWYNTN-UHFFFAOYSA-M copper(I) chloride Chemical compound [Cu]Cl OXBLHERUFWYNTN-UHFFFAOYSA-M 0.000 claims description 3
- 229940045803 cuprous chloride Drugs 0.000 claims description 3
- 230000007935 neutral effect Effects 0.000 claims description 3
- 238000010025 steaming Methods 0.000 claims description 3
- 230000006837 decompression Effects 0.000 claims description 2
- 238000000227 grinding Methods 0.000 claims description 2
- 238000009987 spinning Methods 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims 1
- 230000001376 precipitating effect Effects 0.000 claims 1
- 239000000463 material Substances 0.000 abstract description 24
- 230000008929 regeneration Effects 0.000 abstract description 8
- 238000011069 regeneration method Methods 0.000 abstract description 8
- 230000005012 migration Effects 0.000 abstract 1
- 238000013508 migration Methods 0.000 abstract 1
- 229960000583 acetic acid Drugs 0.000 description 12
- 208000027418 Wounds and injury Diseases 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 7
- 210000001188 articular cartilage Anatomy 0.000 description 7
- 208000014674 injury Diseases 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 206010007710 Cartilage injury Diseases 0.000 description 6
- 230000006378 damage Effects 0.000 description 6
- 238000001523 electrospinning Methods 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 5
- 230000007547 defect Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 238000007151 ring opening polymerisation reaction Methods 0.000 description 5
- 238000002390 rotary evaporation Methods 0.000 description 5
- 241001465754 Metazoa Species 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 229910021645 metal ion Inorganic materials 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 3
- 230000000844 anti-bacterial effect Effects 0.000 description 3
- 210000004204 blood vessel Anatomy 0.000 description 3
- 210000001612 chondrocyte Anatomy 0.000 description 3
- 238000007796 conventional method Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- BXYVQNNEFZOBOZ-UHFFFAOYSA-N n-[3-(dimethylamino)propyl]-n',n'-dimethylpropane-1,3-diamine Chemical compound CN(C)CCCNCCCN(C)C BXYVQNNEFZOBOZ-UHFFFAOYSA-N 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 2
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 2
- 208000002804 Osteochondritis Diseases 0.000 description 2
- 201000009859 Osteochondrosis Diseases 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 239000012620 biological material Substances 0.000 description 2
- FPCJKVGGYOAWIZ-UHFFFAOYSA-N butan-1-ol;titanium Chemical compound [Ti].CCCCO.CCCCO.CCCCO.CCCCO FPCJKVGGYOAWIZ-UHFFFAOYSA-N 0.000 description 2
- 230000021164 cell adhesion Effects 0.000 description 2
- 210000002808 connective tissue Anatomy 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- RRAFCDWBNXTKKO-UHFFFAOYSA-N eugenol Chemical compound COC1=CC(CC=C)=CC=C1O RRAFCDWBNXTKKO-UHFFFAOYSA-N 0.000 description 2
- 210000002744 extracellular matrix Anatomy 0.000 description 2
- 210000003035 hyaline cartilage Anatomy 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 150000002500 ions Chemical group 0.000 description 2
- 239000010410 layer Substances 0.000 description 2
- 229910052751 metal Inorganic materials 0.000 description 2
- 239000002184 metal Substances 0.000 description 2
- 210000005036 nerve Anatomy 0.000 description 2
- 230000036407 pain Effects 0.000 description 2
- 239000002861 polymer material Substances 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 229920001059 synthetic polymer Polymers 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 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 1
- 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 1
- OEOIWYCWCDBOPA-UHFFFAOYSA-N 6-methyl-heptanoic acid Chemical compound CC(C)CCCCC(O)=O OEOIWYCWCDBOPA-UHFFFAOYSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- NPBVQXIMTZKSBA-UHFFFAOYSA-N Chavibetol Natural products COC1=CC=C(CC=C)C=C1O NPBVQXIMTZKSBA-UHFFFAOYSA-N 0.000 description 1
- 206010061762 Chondropathy Diseases 0.000 description 1
- 229920001634 Copolyester Polymers 0.000 description 1
- 208000012661 Dyskinesia Diseases 0.000 description 1
- 239000005770 Eugenol Substances 0.000 description 1
- 102000055008 Matrilin Proteins Human genes 0.000 description 1
- 108010072582 Matrilin Proteins Proteins 0.000 description 1
- 208000016285 Movement disease Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- QGMRQYFBGABWDR-UHFFFAOYSA-M Pentobarbital sodium Chemical compound [Na+].CCCC(C)C1(CC)C(=O)NC(=O)[N-]C1=O QGMRQYFBGABWDR-UHFFFAOYSA-M 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- UVMRYBDEERADNV-UHFFFAOYSA-N Pseudoeugenol Natural products COC1=CC(C(C)=C)=CC=C1O UVMRYBDEERADNV-UHFFFAOYSA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 208000007536 Thrombosis Diseases 0.000 description 1
- RHQDFWAXVIIEBN-UHFFFAOYSA-N Trifluoroethanol Chemical compound OCC(F)(F)F RHQDFWAXVIIEBN-UHFFFAOYSA-N 0.000 description 1
- 238000005299 abrasion Methods 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000002473 artificial blood Substances 0.000 description 1
- 239000012237 artificial material Substances 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229960005188 collagen Drugs 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 239000012141 concentrate Substances 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
- 230000006196 deacetylation Effects 0.000 description 1
- 238000003381 deacetylation reaction Methods 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000002049 effect on nutrition Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229960002217 eugenol Drugs 0.000 description 1
- 210000003414 extremity Anatomy 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 229940014259 gelatin Drugs 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 229920002674 hyaluronan Polymers 0.000 description 1
- 229960003160 hyaluronic acid Drugs 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 230000002427 irreversible effect Effects 0.000 description 1
- 210000003041 ligament Anatomy 0.000 description 1
- 210000003407 lower extremity of femur Anatomy 0.000 description 1
- 238000005461 lubrication Methods 0.000 description 1
- 210000002751 lymph Anatomy 0.000 description 1
- 210000001365 lymphatic vessel Anatomy 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000002121 nanofiber Substances 0.000 description 1
- 239000005445 natural material Substances 0.000 description 1
- 238000011587 new zealand white rabbit Methods 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 230000000399 orthopedic effect Effects 0.000 description 1
- 201000008482 osteoarthritis Diseases 0.000 description 1
- 210000004417 patella Anatomy 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 229960002275 pentobarbital sodium Drugs 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 235000010413 sodium alginate Nutrition 0.000 description 1
- 229940005550 sodium alginate Drugs 0.000 description 1
- 239000000661 sodium alginate Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 210000005065 subchondral bone plate Anatomy 0.000 description 1
- 238000004154 testing of material Methods 0.000 description 1
- 230000008733 trauma Effects 0.000 description 1
- 230000000472 traumatic effect Effects 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/26—Mixtures of macromolecular compounds
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
-
- 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
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/56—Porous materials, e.g. foams or sponges
-
- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0015—Electro-spinning characterised by the initial state of the material
- D01D5/003—Electro-spinning characterised by the initial state of the material the material being a polymer solution or dispersion
-
- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
- D01D5/0076—Electro-spinning characterised by the electro-spinning apparatus characterised by the collecting device, e.g. drum, wheel, endless belt, plate or grid
- D01D5/0084—Coating by electro-spinning, i.e. the electro-spun fibres are not removed from the collecting device but remain integral with it, e.g. coating of prostheses
-
- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/40—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties
- D04H1/42—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres from fleeces or layers composed of fibres without existing or potential cohesive properties characterised by the use of certain kinds of fibres insofar as this use has no preponderant influence on the consolidation of the fleece
- D04H1/4326—Condensation or reaction polymers
- D04H1/435—Polyesters
-
- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/70—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres
- D04H1/72—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged
- D04H1/728—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged by electro-spinning
-
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/20—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices containing or releasing organic materials
- A61L2300/252—Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines
-
- 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
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
-
- 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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/06—Materials or treatment for tissue regeneration for cartilage reconstruction, e.g. meniscus
-
- 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
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Abstract
本发明公开了一种软骨修复用明胶聚己内酯蛋白肽复合气凝胶及制备方法,所述软骨修复用明胶聚己内酯蛋白肽复合气凝胶具有良好的生物相容性、可降解性,明胶、聚己内酯静电纺丝材料的引入改变了胶原蛋白肽的原有形态,形成了特殊的纳米网状结构,提高了其物质交换能力,增强了其对细胞的迁移及黏附、组织的新生,促进了软骨组织的再生。
Description
技术领域
本发明涉及生物医学材料技术领域,尤其涉及一种软骨修复用明胶聚己内酯蛋白肽复合气凝胶及制备方法。
背景技术
关节软骨位于活动关节的表面,在负荷的传导和吸收方面发挥重要的作用。在机体运动过程中关节软骨起到负荷传递、震动吸收、抗磨损和润滑的作用。人的一生中社会活动都离不开关节软骨的正常功能。在骨科领域中,关节软骨缺损是十分常见的疾病。关节软骨是无血管、淋巴管、神经的单一结缔组织,自身修复的能力有限,损伤后很难再生。损伤后会产生关节区域的疼痛、关节运动障碍。关节软骨损伤是由外伤或关节退变造成了软骨结构破坏性疾病,是造成人活动障碍和疼痛的主要原因,其发病率也随着居民的运动需求而逐年增长。
由于关节软骨多位于负重关节,且损伤后再生修复能力较差,一旦发生损伤常进展为不可逆的局部病理改变,并且有形成创伤性骨软骨炎、剥脱性骨软骨炎甚至骨性关节炎的可能,严重影响患者的生活质量。关节软骨主要成分为透明软骨和细胞外基质,其中透明软骨数量较少,再生能力较差,但其对软骨组织的营养和再生有重要调节作用,然而由于软骨组织内缺乏血管、神经和淋巴组织,其损伤后自我修复能力较差,难以在原位形成足够力学强度的软骨组织缓冲人体应力。因此,软骨修复是目前治疗软骨损伤和恢复局部力学环境的唯一方法。
目前用于软骨组织工程材料主要分为天然的和人工合成两种,天然来源的材料主要有海藻酸钠、明胶、透明质酸、胶原等;人工合成的材料主要有聚乙二醇、丙交酯乙交酯共聚酯、聚己内酯、聚乳酸、聚乳酸-羟基乙酸等。天然来源的材料含有许多细胞结合位点以及生物分子信号,有利于细胞的粘附、增长及分化;但是生物降解太快,机械性能及可电纺性较差。合成高分子材料虽然机械性能好,容易电纺成丝,但没有细胞亲合位点。单一的凝胶材料的修复效果较弱,并且还存在生物力学强度较差、机械性能不足等缺陷。为了实现二者的优势互补,将天然材料和合成高分子材料混合电纺成复合纳米纤维已广泛应用于制备各种仿生和具有较好生物相容性的组织工程支架。
CN107217388A公开了一种抗菌性聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶电纺纤维膜及制备方法,该电纺纤维膜是由直径为200-1200nm的聚(ε-己内酯)/聚(ε-己内酯)-REDV/明胶纤维和包载在纤维内部的植物源抗菌剂构成的,厚度为50-150μm。制备方法是将聚(ε-己内酯)、聚(ε-己内酯)-REDV和明胶溶于三氟乙醇中,并滴加冰醋酸至溶液澄清,形成电纺溶液,然后加入植物源抗菌剂丁香酚,使用单道注射泵电纺装置,采用静电纺丝方法得到电纺纤维膜。制得的电纺纤维膜具有加快材料表面内皮化与抗菌的双重功能,在人工血管生物医用材料方面具有应用前景。但是该发明所用聚己内酯主要是通过钛酸丁酯、异辛酸亚锡等金属配合物开环聚合而成,在后续加工中这些有毒的金属离子难以完全去除,而且其细胞亲合性差,植入体内后易引发排异反应,另外该材料抗压强度还有待加强,软骨修复性能还有待提高。
发明内容
有鉴于现有技术的上述缺陷,本发明所要解决的技术问题是提供一种抗压强度和软骨修复性能好的软骨修复用明胶聚己内酯蛋白肽复合气凝胶。
为了实现上述发明目的,本发明采用了如下的技术方案:
一种软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,包括如下步骤:
S1将明胶、聚己内酯加入到六氟异丙醇中搅拌,加入乙酸水溶液继续搅拌,然后制备明胶、聚己内酯静电纺丝膜,将静电纺丝膜在冷冻干燥后经紫外线照射备用;
S2将牛骨粉加水提取,收集提取液过滤得到滤液,再将滤液浓缩得到浓缩液,然后向浓缩液中添加胰蛋白酶,经酶解、灭酶、离心后取上清液,将上清液过滤,将所得滤液超滤处理后经紫外线照射得到牛骨胶原蛋白肽溶液备用;
S3将步骤S1得到的静电纺丝膜在研磨仪中制备成短纤维后加入到叔丁醇水溶液中得到短纤维叔丁醇溶液,然后将短纤维叔丁醇溶液与牛骨胶原蛋白肽溶液混合均匀后经冷冻干燥得到软骨修复用明胶聚己内酯蛋白肽复合气凝胶。
优选的,所述软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,包括如下步骤:
S1将明胶、聚己内酯加入到200-300mL六氟异丙醇中,在30-50℃下以600-800rpm搅拌速度搅拌20-30min,加入10-20mL 0-5℃0.1-0.5wt%的乙酸水溶液,在30-50℃下以600-800rpm搅拌速度继续搅拌20-24h,然后在固定电压10-30KV,流速为1-2mL/h条件下制备明胶、聚己内酯静电纺丝膜,纺丝膜的厚度为20nm-10μm,将静电纺丝膜在-25~-20℃下冷冻干燥8-10h后经紫外线照射20-30min后备用;
S2将牛骨粉与水混合后在95-100℃下提取8-10h,提取2-3次,合并提取液过滤得到滤液,再将滤液于55-60℃下旋蒸至提取液体积的20-30%得到浓缩液,然后添加胰蛋白酶,在35-38℃下酶解3-4h,然后将该酶解液放入高压灭菌锅中,98-100℃灭酶10-15min,待其冷却后于6500-8000rpm下离心10-12min取上清液,将上清液过滤,将所得滤液液用<3KDa的超滤膜包超滤处理后经紫外线照射20-30min得到牛骨胶原蛋白肽溶液备用;
S3将步骤S1得到的静电纺丝膜在高通量组织研磨仪中制备成1μm~1mm短纤维后加入到80-90wt%叔丁醇水溶液中得到短纤维叔丁醇溶液,然后将短纤维叔丁醇溶液与牛骨胶原蛋白肽溶液混合均匀后,在﹣50~﹣40℃下冷冻干燥20-24h得到软骨修复用明胶聚己内酯蛋白肽复合气凝胶。
优选的,所述步骤S1中明胶、聚己内酯、六氟异丙醇、乙酸水溶液的质量比为5-10:2-5:15-25:3-10。
优选的,所述步骤S2中牛骨粉与水的质量比为1:10-15。
优选的,所述步骤S2中胰蛋白酶的加入量为浓缩液质量的0.1-0.5wt%。
优选的,所述步骤S3中静电纺丝膜、叔丁醇水溶液的质量比为1-3:5-10,短纤维叔丁醇溶液与牛骨胶原蛋白肽溶液的质量比为3-5:1。
进一步优选的,所述聚己内酯为改性聚己内酯,其制备方法如下:
1)将壳聚糖加入到10-20wt%乙酸水溶液中,加入乙酸酐,加热至80-120℃,反应4-6h,冷却至室温,加入0-5℃水,搅拌5-10min后用氯仿萃取2-3次,合并有机相,有机相用无水硫酸钠干燥10-12h,过滤,收集滤液,将滤液减压蒸除溶剂后得到的剩余物在60-80℃干燥6-8h得到酰化壳聚糖;
2)在氮气气氛下,将步骤1)得到的酰化壳聚糖、苄胺、四氢呋喃混合,在50-70℃反应3-5h,冷却至室温,减压浓缩至原体积的20-30%后,用氯仿萃取2-3次,合并有机层,并用无水硫酸钠干燥10-12h,过滤,收集滤液后减压蒸除溶剂,将剩余物经柱层析纯化,流动相为体积比为2-3:1的乙酸乙酯与正己烷的混合,将洗脱液减压蒸除溶剂后得到的剩余物在60-80℃干燥4-6h得到乙酰壳聚糖衍生物;
3)在氮气气氛下,将步骤2)得到的乙酰壳聚糖衍生物、四氢呋喃、三乙胺混合后,在0-5℃下以1-2滴/秒的速度滴加甲基丙烯酰氯,滴加完成后,在0-5℃下继续反应4-6h,反应完成后减压浓缩至原体积的20-30%后,将剩余液体用氯仿萃取2-3次,合并有机层,并用无水硫酸钠干燥10-12h,过滤,收集滤液后减压蒸除溶剂,将剩余物经柱层析纯化,流动相为体积比为2-3:1的乙酸乙酯与正己烷的混合,将洗脱液减压蒸除溶剂后得到的固体在60-80℃干燥4-6h得到酯化壳聚糖衍生物;
4)在氩气气氛下,将步骤3)得到的酯化壳聚糖衍生物加入到N,N-二甲基甲酰胺中于室温下搅拌10-20min,加入四甲基二丙烯三胺、α-溴异丁酰溴,在室温下继续搅拌10-20min,加入氯化亚铜混合均匀后加热至80-120℃,反应18-20h,反应完成后向反应液中加入0-5℃的无水甲醇,搅拌5-10min,有沉淀产生,过滤,收集沉淀,将沉淀用四氢呋喃溶解后通过中性Al2O3柱,收集滤液,滤液减压蒸除溶剂后得到的剩余物在60-80℃干燥4-6h得到壳聚糖基偶合物;
5)将ε-己内酯、壳聚糖基偶合物、乳糖酶、分子筛混合后在40-60℃搅拌20-30min后加入二氯甲烷,在40-60℃继续搅拌1-2h后,冷却至室温,过滤,收集滤液,将滤液减压蒸除溶剂后加入到0-5℃的无水甲醇中,在6000-8000rpm转速下离心1-2h,过滤,收集沉淀物,沉淀物用丙酮抽提20-24h后在30-50℃下干燥40-48h得到改性聚己内酯。
优选的,所述步骤1)中壳聚糖、乙酸水溶液、乙酸酐的用量比为2-5g:50-100mL:20-30mL。
优选的,所述步骤2)中酰化壳聚糖、苄胺、四氢呋喃的用量比为1-3g:3-5g:30-50mL。
优选的,所述步骤3)中乙酰壳聚糖衍生物、四氢呋喃、三乙胺、甲基丙烯酰氯的用量比为15-20g:200-300mL:4-8g:500-800g。
优选的,所述步骤4)中酯化壳聚糖衍生物、N,N-二甲基甲酰胺、四甲基二丙烯三胺、α-溴异丁酰溴、氯化亚铜、无水甲醇的用量比为4-6g:200-300mL:0.5-1.5g:0.05-0.2g:0.1-0.5g:50-100mL。
优选的,所述步骤5)中ε-己内酯、壳聚糖基偶合物、乳糖酶、分子筛、二氯甲烷、无水甲醇的用量比为5-10:3-5:0.1-0.5:10-20:100-200mL:50-100mL。
明胶是最早应用于软骨组织工程的天然生物材料之一,具有良好的生物相容性,但因其难以达到足够的机械强度难以直接用于软骨修复;聚己内酯则能够弥补明胶在机械强度方面的缺陷,胶原蛋白肽是生物体结缔组织的主要组成成分,具有良好的生物相容性,其在很大程度上保存了天然软骨基质并包含一定量的生长因子,可以很好的弥补生物材料在抗压强度上的缺陷,本发明明胶、聚己内酯静电纺丝材料的引入改变了胶原蛋白肽的原有形态,形成了特殊的纳米网状结构,提高了其物质交换能力,增强了其对细胞的迁移及黏附、组织的新生,促进了软骨组织的再生。
发明人通过静电纺丝技术制备出的明胶聚己内酯胶原蛋白肽复合气凝胶具有很好的吸附能力和超高的多孔性,能模拟软骨细胞外基质从而促进软骨损伤修复。但是聚己内酯主要是通过钛酸丁酯、异辛酸亚锡等金属配合物开环聚合而成,在后续加工中这些有毒的金属离子难以完全去除,虽然其容易电纺成丝,但其分子上没有细胞亲合位点,细胞亲合性差,植入体内后易引发排异反应,从而引起无菌性炎症。发明人利用乳糖酶促进开环聚合反应将含有大量反应型基团的壳聚糖基偶合物枝连上聚己内酯上,对聚己内酯的末端进行官能化改性,大大增加了聚己内酯分子上的细胞亲合位点,从而提高了其细胞亲合性,避免了常规方法制备聚己内酯过程中金属离子的残留;该壳聚糖基偶合物的制备首先是通过壳聚糖在醋酸钠的存在下与乙酸酐反应得到酰化壳聚糖,然后在苄胺的存在下反应得到乙酰壳聚糖衍生物,乙酰壳聚糖衍生物与甲基丙烯酰氯反应得到酯化壳聚糖衍生物,最后酯化壳聚糖衍生物与四甲基二丙烯三胺经过偶联得到该壳聚糖基偶合物。相较于壳聚糖,该壳聚糖基偶合物具有更多的活性基团,能更好的提高聚己内酯的水溶性和细胞黏附性,并且能更好地与明胶进行交联,提高了材料的机械强度,增加了材料的孔洞,经静电纺丝后能更好的负载胶原蛋白肽,为软骨细胞的黏附及生长提供稳定的环境,促进胶原的再生,从而更好的加速软骨损伤的修复,并且枝连上的壳聚糖基提高了材料的抗菌性能,有利于炎症的抑制。
与现有技术相比,本发明具有的有益效果:本发明制备的于软骨修复的电纺明胶聚己内酯胶原蛋白肽复合气凝胶具有良好的生物相容性、可降解性;发明人利用乳糖酶促进开环聚合反应将含有大量反应型基团的壳聚糖基偶合物枝连上聚己内酯上,增加了聚己内酯分子上的细胞亲合位点,避免了常规方法制备聚己内酯过程中金属离子的残留;提高了聚己内酯的水溶性和细胞黏附性,并且能更好地与明胶进行交联,提高了材料的机械强度,增加了材料的孔洞,经静电纺丝后能更好的负载胶原蛋白肽,为软骨细胞的黏附及生长提供稳定的环境,促进胶原的再生,从而更好的加速软骨损伤的修复。
具体实施方式
为免赘述,以下实施例中用到的物品若无特别说明则均市售产品,用到的方法若无特别说明则均为常规方法。
本发明所用部分原料来源如下:
聚己内酯,含水量为0.35%,熔点为58-60℃,,瑞典Perstorp 6500。
牛骨粉,目数为80目,含量为98%。
胰蛋白酶,酶活力≧4万,有效温度范围为10-60℃。
壳聚糖,脱乙酰度≥95%,粘度≤500mpa·s,水分含量≤8%。
乳糖酶,酶活力为1万,酶活力保存率为99.99%。
分子筛,直径为2-3mm,抗压强度≥40N,堆积密度≥0.6g/mL,型号为5A。
实施例1
一种软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,包括如下步骤:
S1将8.8g明胶、4.3g聚己内酯加入到250mL六氟异丙醇中,在40℃下以700rpm搅拌速度搅拌25min,加入6.5g 3℃0.2wt%的乙酸水溶液,在40℃下以700rpm搅拌速度继续搅拌24h,然后在固定电压20KV,流速为2mL/h条件下制备明胶、聚己内酯静电纺丝膜,纺丝膜的厚度为1μm,将静电纺丝膜在-20℃下冷冻干燥8h后经波长为365nm的紫外线照射30min后备用;
S2将10g牛骨粉与120g水混合后在100℃下提取10h,提取3次,合并提取液过滤得到滤液,再将滤液于55℃下旋蒸至提取液体积的20%得到浓缩液,然后添加胰蛋白酶,胰蛋白酶的添加量为浓缩液质量的0.2wt%,在36℃下酶解4h,然后将该酶解液放入高压灭菌锅中,98℃灭酶10min,待其冷却后于7000rpm下离心10min取上清液,将上清液过滤,将所得滤液液用<3KDa的超滤膜包超滤处理后经波长为365nm的紫外线照射30min得到牛骨胶原蛋白肽溶液备用;
S3将10.5g步骤S1得到的静电纺丝膜在高通量组织研磨仪中制备成1μm~1mm短纤维后加入到100mL 90wt%叔丁醇水溶液中得到短纤维叔丁醇溶液,然后将短纤维叔丁醇溶液与30g牛骨胶原蛋白肽溶液混合均匀后,在﹣40℃下冷冻干燥24h得到软骨修复用明胶聚己内酯蛋白肽复合气凝胶。
所述聚己内酯为改性聚己内酯,其制备方法如下:
1)将30g壳聚糖加入到800mL 15wt%乙酸水溶液中,加入250mL乙酸酐,加热至100℃反应5h,冷却至室温,加入400mL 3℃水,搅拌10min后用氯仿萃取3次,合并有机相,有机相用无水硫酸钠干燥12h,过滤,收集滤液,将滤液减压蒸除溶剂后得到的剩余物在60℃干燥8h得到酰化壳聚糖;
2)在氮气气氛下,将步骤1)得到的20g酰化壳聚糖、40g苄胺、500mL四氢呋喃混合,在60℃反应4h,冷却至室温,减压浓缩至原体积的20%后,用氯仿萃取3次,合并有机层,并用无水硫酸钠干燥12h,过滤,收集滤液后减压蒸除溶剂,将剩余物经柱层析纯化,流动相为体积比为3:1的乙酸乙酯与正己烷的混合,将洗脱液减压蒸除溶剂后得到的剩余物在60℃干燥6h得到乙酰壳聚糖衍生物;
3)在氮气气氛下,将15g步骤2)得到的乙酰壳聚糖衍生物、300mL四氢呋喃、5g三乙胺混合后,在5℃下以2滴/秒的速度滴加500g甲基丙烯酰氯,滴加完成后,在5℃下继续反应5h,反应完成后减压浓缩至原体积的20%后,将剩余液体用氯仿萃取3次,合并有机层,并用无水硫酸钠干燥12h,过滤,收集滤液后减压蒸除溶剂,将剩余物经柱层析纯化,流动相为体积比为3:1的乙酸乙酯与正己烷的混合,将洗脱液减压蒸除溶剂后得到的固体在80℃干燥6h得到酯化壳聚糖衍生物;
4)在氩气气氛下,将6g步骤3)得到的酯化壳聚糖衍生物加入到300mL N,N-二甲基甲酰胺中于室温下搅拌15min,加入1.2g四甲基二丙烯三胺、0.15gα-溴异丁酰溴,在室温下继续搅拌20min,加入0.5g氯化亚铜混合均匀后加热至100℃反应20h,反应完成后向反应液中加入100mL 5℃的无水甲醇,搅拌10min,有沉淀产生,过滤,收集沉淀,将沉淀用四氢呋喃溶解后通过中性Al2O3柱,收集滤液,滤液减压蒸除溶剂后得到的剩余物在80℃干燥6h得到壳聚糖基偶合物;
5)将10gε-己内酯、4g壳聚糖基偶合物、0.3g乳糖酶、20g 5A分子筛混合后在50℃搅拌30min后加入200mL二氯甲烷,在50℃继续搅拌2h后,冷却至室温,过滤,收集滤液,将滤液减压蒸除溶剂后加入到5℃的100mL无水甲醇中,在7000rpm转速下离心2h,过滤,收集沉淀物,沉淀物用丙酮抽提24h后在40℃下干燥48h得到改性聚己内酯。
对比例1
一种软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,包括如下步骤:
S1将8.8g明胶、4.3g聚己内酯加入到250mL六氟异丙醇中,在40℃下以700rpm搅拌速度搅拌25min,加入6.5g 3℃0.2wt%的乙酸水溶液,在40℃下以700rpm搅拌速度继续搅拌24h,然后在固定电压20KV,流速为2mL/h条件下制备明胶、聚己内酯静电纺丝膜,纺丝膜的厚度为1μm,将静电纺丝膜在-20℃下冷冻干燥8h后经波长为365nm的紫外线照射30min后备用;
S2将10g牛骨粉与120g水混合后在100℃下提取10h,提取3次,合并提取液过滤得到滤液,再将滤液于55℃下旋蒸至提取液体积的20%得到浓缩液,然后添加胰蛋白酶,胰蛋白酶的添加量为浓缩液质量的0.2wt%,在36℃下酶解4h,然后将该酶解液放入高压灭菌锅中,98℃灭酶10min,待其冷却后于7000rpm下离心10min取上清液,将上清液过滤,将所得滤液液用<3KDa的超滤膜包超滤处理后经波长为365nm的外线照射30min得到牛骨胶原蛋白肽溶液备用;
S3将10.5g步骤S1得到的静电纺丝膜在高通量组织研磨仪中制备成1μm~1mm短纤维后加入到100mL 90wt%叔丁醇水溶液中得到短纤维叔丁醇溶液,然后将短纤维叔丁醇溶液与30g牛骨胶原蛋白肽溶液混合均匀后,在﹣40℃下冷冻干燥24h得到软骨修复用明胶聚己内酯蛋白肽复合气凝胶。
所述聚己内酯为改性聚己内酯,其制备方法如下:
1)将30g壳聚糖加入到800mL 15wt%乙酸水溶液中,加入250mL乙酸酐,加热至100℃反应5h,冷却至室温,加入400mL 3℃水,搅拌10min后用氯仿萃取3次,合并有机相,有机相用无水硫酸钠干燥12h,过滤,收集滤液,将滤液减压蒸除溶剂后得到的剩余物在60℃干燥8h得到酰化壳聚糖;
2)在氮气气氛下,将步骤1)得到的20g酰化壳聚糖、40g苄胺、500mL四氢呋喃混合,在60℃反应4h,冷却至室温,减压浓缩至原体积的20%后,用氯仿萃取3次,合并有机层,并用无水硫酸钠干燥12h,过滤,收集滤液后减压蒸除溶剂,将剩余物经柱层析纯化,流动相为体积比为3:1的乙酸乙酯与正己烷的混合,将洗脱液减压蒸除溶剂后得到的剩余物在60℃干燥6h得到乙酰壳聚糖衍生物;
3)在氮气气氛下,将15g步骤2)得到的乙酰壳聚糖衍生物、300mL四氢呋喃、5g三乙胺混合后,在5℃下以2滴/秒的速度滴加500g甲基丙烯酰氯,滴加完成后,在5℃下继续反应5h,反应完成后减压浓缩至原体积的20%后,将剩余液体用氯仿萃取3次,合并有机层,并用无水硫酸钠干燥12h,过滤,收集滤液后减压蒸除溶剂,将剩余物经柱层析纯化,流动相为体积比为3:1的乙酸乙酯与正己烷的混合,将洗脱液减压蒸除溶剂后得到的固体在80℃干燥6h得到酯化壳聚糖衍生物;
4)将10gε-己内酯、4g酯化壳聚糖衍生物、0.3g乳糖酶、20g 5A分子筛混合后在50℃搅拌30min后加入200mL二氯甲烷,在50℃继续搅拌2h后,冷却至室温,过滤,收集滤液,将滤液减压蒸除溶剂后加入到5℃的100mL无水甲醇中,在7000rpm转速下离心2h,过滤,收集沉淀物,沉淀物用丙酮抽提24h后在40℃下干燥48h得到改性聚己内酯。
对比例2
一种软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,包括如下步骤:
S1将8.8g明胶、4.3g聚己内酯加入到250mL六氟异丙醇中,在40℃下以700rpm搅拌速度搅拌25min,加入6.5g 3℃0.2wt%的乙酸水溶液,在40℃下以700rpm搅拌速度继续搅拌24h,然后在固定电压20KV,流速为2mL/h条件下制备明胶、聚己内酯静电纺丝膜,纺丝膜的厚度为1μm,将静电纺丝膜在-20℃下冷冻干燥8h后经波长为365nm的紫外线照射30min后备用;
S2将10g牛骨粉与120g水混合后在100℃下提取10h,提取3次,合并提取液过滤得到滤液,再将滤液于55℃下旋蒸至提取液体积的20%得到浓缩液,然后添加胰蛋白酶,胰蛋白酶的添加量为浓缩液质量的0.2wt%,在36℃下酶解4h,然后将该酶解液放入高压灭菌锅中,98℃灭酶10min,待其冷却后于7000rpm下离心10min取上清液,将上清液过滤,将所得滤液液用<3KDa的超滤膜包超滤处理后经波长为365nm的紫外线照射30min得到牛骨胶原蛋白肽溶液备用;
S3将10.5g步骤S1得到的静电纺丝膜在高通量组织研磨仪中制备成1μm~1mm短纤维后加入到100mL 90wt%叔丁醇水溶液中得到短纤维叔丁醇溶液,然后将短纤维叔丁醇溶液与30g牛骨胶原蛋白肽溶液混合均匀后,在﹣40℃下冷冻干燥24h得到软骨修复用明胶聚己内酯蛋白肽复合气凝胶。
所述聚己内酯为改性聚己内酯,其制备方法如下:
将10gε-己内酯、4g壳聚糖、0.3g乳糖酶、20g 5A分子筛混合后在50℃搅拌30min后加入200mL二氯甲烷,在50℃继续搅拌2h后,冷却至室温,过滤,收集滤液,将滤液减压蒸除溶剂后加入到5℃的100mL无水甲醇中,在7000rpm转速下离心2h,过滤,收集沉淀物,沉淀物用丙酮抽提24h后在40℃下干燥48h得到改性聚己内酯。
对比例3
一种软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,包括如下步骤:
S1将8.8g明胶、4.3g聚己内酯加入到250mL六氟异丙醇中,在40℃下以700rpm搅拌速度搅拌25min,加入6.5g 3℃0.2wt%的乙酸水溶液,在40℃下以700rpm搅拌速度继续搅拌24h,然后在固定电压20KV,流速为2mL/h条件下制备明胶、聚己内酯静电纺丝膜,纺丝膜的厚度为1μm,将静电纺丝膜在-20℃下冷冻干燥8h后经波长为365nm的紫外线照射30min后备用;
S2将10g牛骨粉与120g水混合后在100℃下提取10h,提取3次,合并提取液过滤得到滤液,再将滤液于55℃下旋蒸至提取液体积的20%得到浓缩液,然后添加胰蛋白酶,胰蛋白酶的添加量为浓缩液质量的0.2wt%,在36℃下酶解4h,然后将该酶解液放入高压灭菌锅中,98℃灭酶10min,待其冷却后于7000rpm下离心10min取上清液,将上清液过滤,将所得滤液液用<3KDa的超滤膜包超滤处理后经波长为365nm的紫外线照射30min得到牛骨胶原蛋白肽溶液备用;
S3将10.5g步骤S1得到的静电纺丝膜在高通量组织研磨仪中制备成1μm~1mm短纤维后加入到100mL 90wt%叔丁醇水溶液中得到短纤维叔丁醇溶液,然后将短纤维叔丁醇溶液与30g牛骨胶原蛋白肽溶液混合均匀后,在﹣40℃下冷冻干燥24h得到软骨修复用明胶聚己内酯蛋白肽复合气凝胶。
测试例1
抗压强度测试:将软骨修复用明胶聚己内酯蛋白肽复合气凝胶制成高8mm,直径6mm的圆柱体,在材料试验机下以5mm/min的速度压缩至原始厚度的50%。记录支架在50%变形时的应力,得到支架材料的抗压强度,测试结果如表1所示:
孔隙率测试:测试样品分别为实施例1、对比例1-3制得的软骨修复用明胶聚己内酯蛋白肽复合气凝胶,将质量为W的样品浸没在无水乙醇中,待乙醇完全填充软骨修复用明胶聚己内酯蛋白肽复合气凝胶中的孔隙内,然后将软骨修复用明胶聚己内酯蛋白肽复合气凝胶转移至含有已知初始体积(V1)和重量(W1)乙醇的量筒中。记录支架和乙醇在量筒中的重量和体积分别为W2和V2。其孔隙率(ε)计算如下:
ε=(W1-W2-W)/(V1-V2)×100%
表1抗压强度、孔隙率测试结果
从表1的实验数据可以看出,实施例1得到的软骨修复用明胶聚己内酯蛋白肽复合气凝胶的抗压性能最好,而实施例1与其它实施例的区别在于添加了经ε-己内酯、壳聚糖基偶合物、乳糖酶促进开环聚合反应将含有大量反应型基团的壳聚糖基偶合物枝连的聚己内酯,可能的原因是该壳聚糖基偶合物改善了聚己内酯的水溶性,能更好地与明胶进行交联,从而提高了材料的力学性能和孔隙率。
测试例2
动物软骨修复实验:
动物模型的建立:取新西兰大白兔50只,3个月年龄大小,体重1.8~2.4kg,手术前6h禁食禁饮。随机分为5组,每组10只,分A、B、C、D、E组,以2.5%戊巴比妥钠耳缘静脉注射麻醉。麻醉妥当后,妥善固定其四肢,依次切开膑韧带外侧缘,充分暴露股骨下端外侧髁负重区,在其上做一个直径4mm、深度3mm的圆柱形缺损,钻穿软骨下骨,清除缺损内的血凝块后,A、B、C、D组分别置入实施例1、对比例1-3的软骨修复用明胶聚己内酯蛋白肽复合气凝胶,E组做空白对照未植入任何材料。逐层缝合伤口,术后碘伏消毒伤口,纱布包扎后不用外固定,肌注青链霉素双抗预防感染,术后单笼喂养,于16周后分别进行CT检查,观察骨软骨的修复情况,并使用国际软骨修复协会(ICRS)评分标准进行评分,具体评分标准如表2所示:
表2ICRS评分标准
动物软骨修复实验结果如表3所示:
表3动物软骨修复实验结果表
从表3中的实验结果可以看出,实施例1制得的软骨修复用明胶聚己内酯蛋白肽复合气凝胶的软骨恢复能力最好,可能的原因是该壳聚糖基偶合物具有更多的活性基团,能更好的提高聚己内酯的水溶性和细胞黏附性,经静电纺丝后能更好的负载胶原蛋白肽,为软骨细胞的黏附及生长提供稳定的环境,促进胶原的再生,从而更好的加速软骨损伤的修复,并且枝连上的壳聚糖基提高了材料的抗菌性能,有利于炎症的抑制。
Claims (6)
1.一种软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,其特征在于,包括如下步骤:
S1将明胶、聚己内酯加入到200-300mL六氟异丙醇中,在30-50℃下以600-800rpm搅拌速度搅拌20-30min,加入10-20mL 0-5℃ 0.1-0.5wt%的乙酸水溶液,在30-50℃下以600-800rpm搅拌速度继续搅拌20-24h,然后在固定电压10-30KV,流速为1-2mL/h条件下制备明胶、聚己内酯静电纺丝膜,纺丝膜的厚度为20nm-10μm,将静电纺丝膜在-25~-20℃下冷冻干燥8-10h后经紫外线照射20-30min后备用;
S2将牛骨粉与水混合后在95-100℃下提取8-10h,提取2-3次,合并提取液过滤得到滤液,再将滤液于55-60℃下旋蒸至提取液体积的20-30%得到浓缩液,然后添加胰蛋白酶,在35-38℃下酶解3-4h,然后将酶解液放入高压灭菌锅中,98-100℃灭酶10-15min,待其冷却后于6500-8000rpm下离心10-12min取上清液,将上清液过滤,将所得滤液超滤处理后经紫外线照射20-30min得到牛骨胶原蛋白肽溶液备用;
S3将步骤S1得到的静电纺丝膜在高通量组织研磨仪中制备成1μm~1mm短纤维后加入到80-90wt%叔丁醇水溶液中得到短纤维叔丁醇溶液,然后将短纤维叔丁醇溶液与牛骨胶原蛋白肽溶液混合均匀后,在﹣50~﹣40℃下冷冻干燥20-24h得到软骨修复用明胶聚己内酯蛋白肽复合气凝胶;
所述聚己内酯为改性聚己内酯,其制备方法如下:
1)将壳聚糖加入到10-20wt%乙酸水溶液中,加入乙酸酐,加热至80-120℃,反应4-6h,冷却至室温,加入0-5℃水,搅拌5-10min后用氯仿萃取2-3次,合并有机相,有机相用无水硫酸钠干燥10-12h,过滤,收集滤液,将滤液减压蒸除溶剂后得到的剩余物在60-80℃干燥6-8h得到酰化壳聚糖;
2)在氮气气氛下,将步骤1)得到的酰化壳聚糖、苄胺、四氢呋喃混合,在50-70℃反应3-5h,冷却至室温,减压浓缩至原体积的20-30%后,用氯仿萃取2-3次,合并有机层,并用无水硫酸钠干燥10-12h,过滤,收集滤液后减压蒸除溶剂,将剩余物经柱层析纯化,流动相为体积比为2-3:1的乙酸乙酯与正己烷的混合液,将洗脱液减压蒸除溶剂后得到的剩余物在60-80℃干燥4-6h得到乙酰壳聚糖衍生物;
3)在氮气气氛下,将步骤2)得到的乙酰壳聚糖衍生物、四氢呋喃、三乙胺混合后,在0-5℃下以1-2滴/秒的速度滴加甲基丙烯酰氯,滴加完成后,在0-5℃下继续反应4-6h,反应完成后减压浓缩至原体积的20-30%后,将剩余液体用氯仿萃取2-3次,合并有机层,并用无水硫酸钠干燥10-12h,过滤,收集滤液后减压蒸除溶剂,将剩余物经柱层析纯化,流动相为体积比为2-3:1的乙酸乙酯与正己烷的混合液,将洗脱液减压蒸除溶剂后得到的固体在60-80℃干燥4-6h得到酯化壳聚糖衍生物;
4)在氩气气氛下,将步骤3)得到的酯化壳聚糖衍生物加入到N,N-二甲基甲酰胺中于室温下搅拌10-20min,加入四甲基二丙烯三胺、α-溴异丁酰溴,在室温下继续搅拌10-20min,加入氯化亚铜混合均匀后加热至80-120℃,反应18-20h,反应完成后向反应液中加入0-5℃的无水甲醇,搅拌5-10min,有沉淀产生,过滤,收集沉淀,将沉淀用四氢呋喃溶解后通过中性Al2O3柱,收集滤液,滤液减压蒸除溶剂后得到的剩余物在60-80℃干燥4-6h得到壳聚糖基偶合物;
5)将ε-己内酯、壳聚糖基偶合物、乳糖酶、分子筛混合后在40-60℃搅拌20-30min后加入二氯甲烷,在40-60℃继续搅拌1-2h后,冷却至室温,过滤,收集滤液,将滤液减压蒸除溶剂后加入到0-5℃的无水甲醇中,在6000-8000rpm转速下离心1-2h,过滤,收集沉淀物,沉淀物用丙酮抽提20-24h后在30-50℃下干燥40-48h得到改性聚己内酯。
2.如权利要求1所述的软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,其特征在于:所述步骤S1中明胶、聚己内酯、六氟异丙醇、乙酸水溶液的质量比为5-10:2-5:15-25:3-10。
3.如权利要求1所述的软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,其特征在于:所述步骤S2中牛骨粉与水的质量比为1:10-15。
4.如权利要求1所述的软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,其特征在于:所述步骤S2中胰蛋白酶的加入量为浓缩液质量的0.1-0.5wt%。
5.如权利要求1所述的软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,其特征在于:所述步骤1)中壳聚糖、乙酸水溶液、乙酸酐的用量比为2-5g:50-100mL:20-30mL。
6.如权利要求1所述的软骨修复用明胶聚己内酯蛋白肽复合气凝胶的制备方法,其特征在于:所述步骤2)中酰化壳聚糖、苄胺、四氢呋喃的用量比为1-3g:3-5g:30-50mL。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310254070.9A CN116212111B (zh) | 2023-03-16 | 2023-03-16 | 软骨修复用明胶聚己内酯蛋白肽复合气凝胶及制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202310254070.9A CN116212111B (zh) | 2023-03-16 | 2023-03-16 | 软骨修复用明胶聚己内酯蛋白肽复合气凝胶及制备方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN116212111A CN116212111A (zh) | 2023-06-06 |
CN116212111B true CN116212111B (zh) | 2023-12-22 |
Family
ID=86569428
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202310254070.9A Active CN116212111B (zh) | 2023-03-16 | 2023-03-16 | 软骨修复用明胶聚己内酯蛋白肽复合气凝胶及制备方法 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN116212111B (zh) |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101704906A (zh) * | 2009-11-25 | 2010-05-12 | 华东师范大学 | 一步合成壳聚糖接枝聚己内酯阳离子共聚物的方法及其应用 |
CN101857649A (zh) * | 2010-03-18 | 2010-10-13 | 沈阳工业大学 | 一种壳寡糖接枝聚己内酯热塑性材料的制备方法 |
CN103147225A (zh) * | 2013-02-06 | 2013-06-12 | 东华大学 | 一种蛋白-多糖-聚乳酸聚己内酯血管支架的制备方法 |
CN111849135A (zh) * | 2020-06-23 | 2020-10-30 | 南宁学院 | 一种聚己内酯复合材料及其制备方法 |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP3720515A4 (en) * | 2017-12-08 | 2021-08-25 | Nanofiber Solutions, LLC | ELECTROSPUNNED FIBERS FOR REPAIR AND REGROWTH OF HYALINE CARTILLE |
-
2023
- 2023-03-16 CN CN202310254070.9A patent/CN116212111B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101704906A (zh) * | 2009-11-25 | 2010-05-12 | 华东师范大学 | 一步合成壳聚糖接枝聚己内酯阳离子共聚物的方法及其应用 |
CN101857649A (zh) * | 2010-03-18 | 2010-10-13 | 沈阳工业大学 | 一种壳寡糖接枝聚己内酯热塑性材料的制备方法 |
CN103147225A (zh) * | 2013-02-06 | 2013-06-12 | 东华大学 | 一种蛋白-多糖-聚乳酸聚己内酯血管支架的制备方法 |
CN111849135A (zh) * | 2020-06-23 | 2020-10-30 | 南宁学院 | 一种聚己内酯复合材料及其制备方法 |
Non-Patent Citations (3)
Title |
---|
Poly-ε-caprolactone/gel hybrid scaffolds for cartilage tissue engineering;J. C. Schagemann等;《Journal of Biomedical Materials Research Part A》;第93A卷(第2期);第454-463页 * |
携带骨粉明胶/聚己内酯静电纺纤维膜支架材料的制备及其生物相容性研究;荣冬明;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》(第02期);E080-79 * |
静电纺丝三维纳米纤维气凝胶结合软骨细胞外基质修复兔软骨损伤模型研究;王磊;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》(第03期);E080-119 * |
Also Published As
Publication number | Publication date |
---|---|
CN116212111A (zh) | 2023-06-06 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Zhang et al. | Synthesis and characterization of a degradable composite agarose/HA hydrogel | |
EP3072536B1 (en) | Hydrophilic electrospinning biological composite stent material used for tissue regeneration and preparation method and application thereof | |
DK1917218T3 (en) | COMPOSITIONS OF PARTIALLY DEACETYLATED CHITIN DERIVATIVES | |
CA2861027C (en) | Collagen structure, and method for producing collagen structure | |
CN107441556B (zh) | 一种聚氨基酸封端的组织修补材料及其制备方法 | |
EP2976112B1 (en) | Improvements in and relating to collagen based materials | |
Wang et al. | Enhanced physical and biological properties of chitosan scaffold by silk proteins cross-linking | |
CN111388755A (zh) | 一种可注射型透明质酸/壳聚糖水凝胶及其制备方法 | |
CN113663137A (zh) | 复合生物补片及其制备方法及应用 | |
WO2011064724A1 (en) | Biomimetic composite materials, preparation process thereof and use thereof to produce mono-, bi- or multi -layer structures for the regeneration of bone, cartilaginous and osteocartilaginous tissue | |
ITPD980037A1 (it) | Acido ialuronico solfatato e i suoi derivati legati covalentemente a polimeri sintetici pe la preparazione di biomateriali e per il rivesti | |
CN107397980B (zh) | 一种组织修复膜涂覆用防粘连组合物及其使用方法 | |
CN116212111B (zh) | 软骨修复用明胶聚己内酯蛋白肽复合气凝胶及制备方法 | |
AU4368099A (en) | Biomaterials containing hyaluronic acid derivatives in the form of three-dimensional structures free from cellular components or products thereof for the in vivo regeneration of tissue cells | |
CN113877001A (zh) | 一种注射用丝素蛋白复合凝胶 | |
Wang et al. | Fabrication, characterization and potential application of biodegradable polydopamine-modified scaffolds based on natural macromolecules | |
Kim et al. | Preparation and properties of collagen/modified hyaluronic acid hydrogel for biomedical application | |
CN111821513A (zh) | 一种促进软骨形成的复合水凝胶及其制备方法和应用 | |
CN115429935B (zh) | 一种可注射性的交联硫酸软骨素水凝胶及其制备方法 | |
CN114681668B (zh) | 一种3d打印的硒掺杂羟基磷灰石人工骨结构的制备方法 | |
CN115192776B (zh) | 制备肌腱损伤修复用强韧水凝胶的方法 | |
CN114395164B (zh) | 一种多糖复合凝胶及其制备方法和应用 | |
CN116139344A (zh) | 一种促进成骨细胞生成的骨修复材料及其制备方法 | |
CN113941033B (zh) | 一种双载药纳米纤维水凝胶复合软骨修复系统及其制备方法 | |
KR101582380B1 (ko) | 손상 조직에 대한 세포 전달 및 조직 재생용 생체재료와 제조방법 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |