CN113651974A - Preparation method of photoinduced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing - Google Patents
Preparation method of photoinduced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing Download PDFInfo
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- CN113651974A CN113651974A CN202110802566.6A CN202110802566A CN113651974A CN 113651974 A CN113651974 A CN 113651974A CN 202110802566 A CN202110802566 A CN 202110802566A CN 113651974 A CN113651974 A CN 113651974A
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- gelatin
- silk fibroin
- printing
- solution
- crosslinked hydrogel
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- 108010022355 Fibroins Proteins 0.000 title claims abstract description 173
- 108010010803 Gelatin Proteins 0.000 title claims abstract description 156
- 239000008273 gelatin Substances 0.000 title claims abstract description 156
- 229920000159 gelatin Polymers 0.000 title claims abstract description 156
- 235000019322 gelatine Nutrition 0.000 title claims abstract description 156
- 235000011852 gelatine desserts Nutrition 0.000 title claims abstract description 156
- 239000000017 hydrogel Substances 0.000 title claims abstract description 49
- 238000010146 3D printing Methods 0.000 title claims abstract description 47
- 238000002360 preparation method Methods 0.000 title claims abstract description 35
- 239000000463 material Substances 0.000 claims abstract description 31
- 238000000034 method Methods 0.000 claims abstract description 30
- 239000002243 precursor Substances 0.000 claims abstract description 29
- 238000004132 cross linking Methods 0.000 claims abstract description 15
- 230000008569 process Effects 0.000 claims abstract description 14
- 239000000243 solution Substances 0.000 claims description 88
- -1 transition metal salts Chemical class 0.000 claims description 33
- 239000007788 liquid Substances 0.000 claims description 29
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 238000007639 printing Methods 0.000 claims description 16
- 238000003756 stirring Methods 0.000 claims description 11
- 239000004744 fabric Substances 0.000 claims description 10
- 239000000499 gel Substances 0.000 claims description 10
- 239000002245 particle Substances 0.000 claims description 10
- 238000007605 air drying Methods 0.000 claims description 8
- 229920002678 cellulose Polymers 0.000 claims description 8
- 239000001913 cellulose Substances 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 8
- 239000008367 deionised water Substances 0.000 claims description 8
- 229910021641 deionized water Inorganic materials 0.000 claims description 8
- 238000000502 dialysis Methods 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- 238000005485 electric heating Methods 0.000 claims description 8
- 230000001678 irradiating effect Effects 0.000 claims description 8
- 230000003647 oxidation Effects 0.000 claims description 8
- 238000007254 oxidation reaction Methods 0.000 claims description 8
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 6
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 claims description 6
- VFLDPWHFBUODDF-FCXRPNKRSA-N curcumin Chemical compound C1=C(O)C(OC)=CC(\C=C\C(=O)CC(=O)\C=C\C=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-FCXRPNKRSA-N 0.000 claims description 6
- 238000001125 extrusion Methods 0.000 claims description 5
- 238000012545 processing Methods 0.000 claims description 5
- 230000008961 swelling Effects 0.000 claims description 5
- ROOXNKNUYICQNP-UHFFFAOYSA-N ammonium persulfate Chemical compound [NH4+].[NH4+].[O-]S(=O)(=O)OOS([O-])(=O)=O ROOXNKNUYICQNP-UHFFFAOYSA-N 0.000 claims description 4
- 239000000975 dye Substances 0.000 claims description 4
- SEACYXSIPDVVMV-UHFFFAOYSA-L eosin Y Chemical compound [Na+].[Na+].[O-]C(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C([O-])=C(Br)C=C21 SEACYXSIPDVVMV-UHFFFAOYSA-L 0.000 claims description 4
- 230000000977 initiatory effect Effects 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 239000007800 oxidant agent Substances 0.000 claims description 4
- JRKICGRDRMAZLK-UHFFFAOYSA-L persulfate group Chemical group S(=O)(=O)([O-])OOS(=O)(=O)[O-] JRKICGRDRMAZLK-UHFFFAOYSA-L 0.000 claims description 4
- XXQBEVHPUKOQEO-UHFFFAOYSA-N potassium superoxide Chemical compound [K+].[K+].[O-][O-] XXQBEVHPUKOQEO-UHFFFAOYSA-N 0.000 claims description 4
- 150000003839 salts Chemical class 0.000 claims description 4
- 229910052723 transition metal Inorganic materials 0.000 claims description 4
- LCPVQAHEFVXVKT-UHFFFAOYSA-N 2-(2,4-difluorophenoxy)pyridin-3-amine Chemical compound NC1=CC=CN=C1OC1=CC=C(F)C=C1F LCPVQAHEFVXVKT-UHFFFAOYSA-N 0.000 claims description 3
- FNPNRZLYTIVLNO-UHFFFAOYSA-N Cl(=O)(=O)(=O)O.C1(=C(C(=CC(=C1)C)C)C1C2=CC=CC=C2N(C=2C=CC=CC12)C)C Chemical compound Cl(=O)(=O)(=O)O.C1(=C(C(=CC(=C1)C)C)C1C2=CC=CC=C2N(C=2C=CC=CC12)C)C FNPNRZLYTIVLNO-UHFFFAOYSA-N 0.000 claims description 3
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 claims description 3
- 239000004148 curcumin Substances 0.000 claims description 3
- 235000012754 curcumin Nutrition 0.000 claims description 3
- 229940109262 curcumin Drugs 0.000 claims description 3
- VFLDPWHFBUODDF-UHFFFAOYSA-N diferuloylmethane Natural products C1=C(O)C(OC)=CC(C=CC(=O)CC(=O)C=CC=2C=C(OC)C(O)=CC=2)=C1 VFLDPWHFBUODDF-UHFFFAOYSA-N 0.000 claims description 3
- USHAGKDGDHPEEY-UHFFFAOYSA-L potassium persulfate Chemical compound [K+].[K+].[O-]S(=O)(=O)OOS([O-])(=O)=O USHAGKDGDHPEEY-UHFFFAOYSA-L 0.000 claims description 3
- 239000002151 riboflavin Substances 0.000 claims description 3
- 235000019192 riboflavin Nutrition 0.000 claims description 3
- 229960002477 riboflavin Drugs 0.000 claims description 3
- CHQMHPLRPQMAMX-UHFFFAOYSA-L sodium persulfate Substances [Na+].[Na+].[O-]S(=O)(=O)OOS([O-])(=O)=O CHQMHPLRPQMAMX-UHFFFAOYSA-L 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- RLDWIFDMZUBMGW-UHFFFAOYSA-N 29h,31h-phthalocyanine, disodium salt Chemical compound [Na+].[Na+].C12=CC=CC=C2C(N=C2[N-]C(C3=CC=CC=C32)=N2)=NC1=NC([C]1C=CC=CC1=1)=NC=1N=C1[C]3C=CC=CC3=C2[N-]1 RLDWIFDMZUBMGW-UHFFFAOYSA-N 0.000 claims description 2
- FAHHTVHENMIZIV-UHFFFAOYSA-N Cl(=O)(=O)(=O)O.CN1C=2C=CC=CC2C(C2=CC=CC=C12)C1=CC=CC=C1 Chemical compound Cl(=O)(=O)(=O)O.CN1C=2C=CC=CC2C(C2=CC=CC=C12)C1=CC=CC=C1 FAHHTVHENMIZIV-UHFFFAOYSA-N 0.000 claims description 2
- 229910021578 Iron(III) chloride Inorganic materials 0.000 claims description 2
- BGLGAKMTYHWWKW-UHFFFAOYSA-N acridine yellow Chemical compound [H+].[Cl-].CC1=C(N)C=C2N=C(C=C(C(C)=C3)N)C3=CC2=C1 BGLGAKMTYHWWKW-UHFFFAOYSA-N 0.000 claims description 2
- 229910001870 ammonium persulfate Inorganic materials 0.000 claims description 2
- 125000006269 biphenyl-2-yl group Chemical group [H]C1=C([H])C([H])=C(C([H])=C1[H])C1=C(*)C([H])=C([H])C([H])=C1[H] 0.000 claims description 2
- YVQNEQKKLNWXEM-UHFFFAOYSA-L dichloroplatinum;2-pyridin-2-ylpyridine Chemical compound Cl[Pt]Cl.N1=CC=CC=C1C1=CC=CC=N1 YVQNEQKKLNWXEM-UHFFFAOYSA-L 0.000 claims description 2
- 238000004090 dissolution Methods 0.000 claims description 2
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 claims description 2
- 230000003287 optical effect Effects 0.000 claims description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-M perchlorate Inorganic materials [O-]Cl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-M 0.000 claims description 2
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 claims description 2
- AZJPTIGZZTZIDR-UHFFFAOYSA-L rose bengal Chemical compound [K+].[K+].[O-]C(=O)C1=C(Cl)C(Cl)=C(Cl)C(Cl)=C1C1=C2C=C(I)C(=O)C(I)=C2OC2=C(I)C([O-])=C(I)C=C21 AZJPTIGZZTZIDR-UHFFFAOYSA-L 0.000 claims description 2
- 229960003138 rose bengal sodium Drugs 0.000 claims description 2
- 238000010008 shearing Methods 0.000 claims description 2
- PFUVRDFDKPNGAV-UHFFFAOYSA-N sodium peroxide Chemical compound [Na+].[Na+].[O-][O-] PFUVRDFDKPNGAV-UHFFFAOYSA-N 0.000 claims description 2
- 238000001228 spectrum Methods 0.000 claims description 2
- YNHJECZULSZAQK-UHFFFAOYSA-N tetraphenylporphyrin Chemical compound C1=CC(C(=C2C=CC(N2)=C(C=2C=CC=CC=2)C=2C=CC(N=2)=C(C=2C=CC=CC=2)C2=CC=C3N2)C=2C=CC=CC=2)=NC1=C3C1=CC=CC=C1 YNHJECZULSZAQK-UHFFFAOYSA-N 0.000 claims description 2
- 229910052724 xenon Inorganic materials 0.000 claims description 2
- FHNFHKCVQCLJFQ-UHFFFAOYSA-N xenon atom Chemical compound [Xe] FHNFHKCVQCLJFQ-UHFFFAOYSA-N 0.000 claims description 2
- 230000033116 oxidation-reduction process Effects 0.000 claims 1
- 230000001590 oxidative effect Effects 0.000 claims 1
- 230000004048 modification Effects 0.000 abstract description 15
- 238000012986 modification Methods 0.000 abstract description 15
- 238000005516 engineering process Methods 0.000 abstract description 10
- 238000006243 chemical reaction Methods 0.000 abstract description 9
- 239000003054 catalyst Substances 0.000 abstract description 3
- 239000003431 cross linking reagent Substances 0.000 abstract description 3
- 239000003607 modifier Substances 0.000 abstract description 3
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 abstract description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 abstract description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 abstract description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 abstract description 2
- 230000009471 action Effects 0.000 abstract description 2
- 229920002521 macromolecule Polymers 0.000 abstract description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 abstract description 2
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 abstract description 2
- 150000001413 amino acids Chemical class 0.000 abstract 1
- 238000006552 photochemical reaction Methods 0.000 abstract 1
- 239000002131 composite material Substances 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 3
- 230000014759 maintenance of location Effects 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 230000021164 cell adhesion Effects 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 102000009123 Fibrin Human genes 0.000 description 1
- 108010073385 Fibrin Proteins 0.000 description 1
- BWGVNKXGVNDBDI-UHFFFAOYSA-N Fibrin monomer Chemical compound CNC(=O)CNC(=O)CN BWGVNKXGVNDBDI-UHFFFAOYSA-N 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- DZGWFCGJZKJUFP-UHFFFAOYSA-N Tyramine Natural products NCCC1=CC=C(O)C=C1 DZGWFCGJZKJUFP-UHFFFAOYSA-N 0.000 description 1
- 230000010933 acylation Effects 0.000 description 1
- 238000005917 acylation reaction Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000000975 bioactive effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 238000010382 chemical cross-linking Methods 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 230000000295 complement effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- WHELTKFSBJNBMQ-UHFFFAOYSA-L dichlororuthenium;2-pyridin-2-ylpyridine;hexahydrate Chemical compound O.O.O.O.O.O.[Cl-].[Cl-].[Ru+2].N1=CC=CC=C1C1=CC=CC=N1.N1=CC=CC=C1C1=CC=CC=N1.N1=CC=CC=C1C1=CC=CC=N1 WHELTKFSBJNBMQ-UHFFFAOYSA-L 0.000 description 1
- 239000002612 dispersion medium Substances 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005538 encapsulation Methods 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 229950003499 fibrin Drugs 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 125000005395 methacrylic acid group Chemical group 0.000 description 1
- DCUFMVPCXCSVNP-UHFFFAOYSA-N methacrylic anhydride Chemical compound CC(=C)C(=O)OC(=O)C(C)=C DCUFMVPCXCSVNP-UHFFFAOYSA-N 0.000 description 1
- 229920005615 natural polymer Polymers 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000027756 respiratory electron transport chain Effects 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000006276 transfer reaction Methods 0.000 description 1
- 229960003732 tyramine Drugs 0.000 description 1
- DZGWFCGJZKJUFP-UHFFFAOYSA-O tyraminium Chemical compound [NH3+]CCC1=CC=C(O)C=C1 DZGWFCGJZKJUFP-UHFFFAOYSA-O 0.000 description 1
- 238000002604 ultrasonography Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/02—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques
- C08J3/03—Making solutions, dispersions, lattices or gels by other methods than by solution, emulsion or suspension polymerisation techniques in aqueous media
- C08J3/075—Macromolecular gels
-
- 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
- A61L27/52—Hydrogels or hydrocolloids
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B33—ADDITIVE MANUFACTURING TECHNOLOGY
- B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3-D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3-D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
- B33Y70/00—Materials specially adapted for additive manufacturing
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
- C08J3/246—Intercrosslinking of at least two polymers
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/28—Treatment by wave energy or particle radiation
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2389/00—Characterised by the use of proteins; Derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2489/00—Characterised by the use of proteins; Derivatives thereof
Abstract
The invention discloses a preparation method of photoinduced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing. In order to meet the specific requirements of different 3D printing equipment on rheological properties of reaction precursors, silk fibroin/gelatin blended solutions with different proportions are used as reaction precursors, and under the action of a visible light-induced redox photoinitiation system, efficient and controllable preparation of the silk fibroin/gelatin co-crosslinked hydrogel material is realized. The invention is based on the crosslinking reaction principle of amino acid with a specific side group structure in the photochemical reaction process, and realizes the rapid preparation of the silk fibroin/gelatin co-crosslinked hydrogel material by utilizing the mutual free radical bonding reaction of tyrosine, phenylalanine, tryptophan and the like in the self structure of the silk fibroin and gelatin macromolecules. The technology does not need to carry out photosensitive modification on silk fibroin and gelatin, avoids potential influence of photosensitive modifiers, catalysts, cross-linking agents and the like on the biocompatibility of a system in the modification process, and has the advantages of good biocompatibility, simple process and high cross-linking forming efficiency.
Description
Technical Field
The invention belongs to the technical field of tissue engineering, and particularly relates to a preparation method of photoinduced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing.
Background
The hydrogel is a polymer network system with a three-dimensional space network structure and taking water as a dispersion medium, and the polymer network system can absorb a large amount of water and has the dual properties of solid and liquid. It has received much attention in the field of tissue engineering because of its high water content, good diffusivity to oxygen and nutrients, and structure similar to the natural extracellular matrix.
The silk fibroin is natural polymer fibrin extracted from silk, has good biocompatibility and excellent mechanical strength, but the silk fibroin solution has low viscosity and poor shape retention of a printing structure, and a cell adhesion site is not arranged on a silk fibroin macromolecular chain, so that the application of the silk fibroin in biological 3D printing is limited. Gelatin is obtained by partial hydrolysis of collagen, has a bioactive motif RGD, and can be used for cell adhesion. In addition, the viscosity of the gelatin solution is adjustable, and the printing structure of the gelatin solution can realize good shape retention. The pure gelatin has temperature-sensitive characteristics, can be gelled at low temperature, but has poor mechanical properties, and is difficult to meet the requirements of various biological 3D printing materials on physical and mechanical properties. The silk fibroin and the gelatin are compounded, so that the performance insufficiency of a single material is avoided, the complementary advantages of the two materials can be realized, and the silk fibroin and gelatin compound printing ink has a better application prospect in biological 3D printing.
At present, various published patents and techniques are used for preparing silk fibroin/gelatin composite hydrogel materials. CN201810521171.7 discloses a silk fibroin/gelatin interpenetrating network hydrogel capable of realizing cell encapsulation and a preparation method thereof. According to the method, a gelatin material is modified by methacrylic anhydride, so that the gelatin is endowed with ultraviolet light induction capability, and the stable interpenetrating network hydrogel is prepared under the combined action of ultraviolet light and ultrasound. Although the technology does not need to add an additional chemical cross-linking agent during the later-stage gelling, the modification process of the gelatin still has the problems of complicated process and potential biocompatibility caused by the use of the modifier and the catalyst. CN201910619312.3 discloses a photo-induced double-modified silk fibroin/gelatin composite bio-ink material and a preparation method thereof. The method carries out methacrylic acylation modification on silk fibroin and gelatin components respectively, and then obtains higher crosslinking degree through ultraviolet light initiated crosslinking reaction. The technology still realizes the mutual cross-linking reaction of the silk fibroin and the gelatin component in a photosensitive modification mode, and the potential influence of the photosensitive modification step and the modification process on the biocompatibility of the material cannot be avoided. It is worth noting that the two technologies both adopt ultraviolet light as an irradiation light source, and the short-wave ultraviolet light often has radiation hazard to cells or medicines in the biological materials, which is not beneficial to the practical application in the later period. CN201810822707.9 discloses a silk fibroin/gelatin double-crosslinked hydrogel and a preparation method thereof. The method comprises the steps of carrying out tyramine root grafting modification on gelatin, dissolving the modified gelatin in a silk fibroin solution, adding horseradish peroxidase to initiate covalent crosslinking of the silk fibroin/gelatin, and finally soaking an alcohol solution to obtain the silk fibroin/gelatin hydrogel. The method realizes covalent crosslinking of modified gelatin and silk fibroin by applying a biological enzyme technology, but the biological enzyme technology has low initiation efficiency and long time consumption, and has strict requirements on the environment. In addition, the technology is still based on graft modification of gelatin materials, and the process is relatively complex.
In conclusion, the existing technology for preparing silk fibroin/gelatin co-crosslinked hydrogel generally needs to carry out graft modification on component materials, and has complex process and great technical difficulty. Meanwhile, the crosslinking reaction efficiency and biocompatibility of the silk fibroin/gelatin composite material cannot be considered, so that the development of a preparation method and a technology of the silk fibroin/gelatin co-crosslinked hydrogel which is free of modification, high in efficiency, simple and excellent in biocompatibility is necessary, and the preparation method and the technology have important values for the application of the silk fibroin/gelatin co-crosslinked hydrogel in the field of biological 3D printing.
Disclosure of Invention
The invention aims to provide a preparation method of photoinduced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing, and aims to solve the problems that the prior art is complex in process and long in time consumption when the silk fibroin/gelatin co-crosslinked hydrogel is prepared, and potential biocompatibility exists in modification.
In order to solve the technical problems, the following technical scheme is adopted
A preparation method of photo-induced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing is characterized by comprising the following steps:
(1) preparation of silk fibroin/gelatin blended liquid
Dissolving, dialyzing, centrifuging and concentrating degummed raw silk grey cloth to obtain a silk fibroin solution, adding solid gelatin particles with certain gel strength and specific proportion into the silk fibroin solution with certain molecular weight for swelling for 15-30 min, and stirring at 60 ℃ until the gelatin particles are completely dissolved to obtain a silk fibroin/gelatin blended solution;
(2) preparation of photo-crosslinking silk fibroin/gelatin precursor solution
Adding a certain amount of dissolved photoinitiation system into the silk fibroin/gelatin blended solution prepared in the step (1) to prepare a photocrosslinking silk fibroin/gelatin precursor solution, and storing the solution in the dark for later use;
(3) preparation of 3D printing silk fibroin/gelatin photoinduced co-crosslinked hydrogel material
And (3) placing the photo-crosslinked silk fibroin/gelatin precursor liquid obtained in the step (2) in a liquid storage tank/pipe of a 3D printing device, adjusting the rheological property of the silk fibroin/gelatin blended liquid through temperature, then printing by using the 3D printing device to obtain a preset structure, and irradiating and crosslinking through visible light with certain intensity in the printing process/after printing to obtain the silk fibroin/gelatin co-crosslinked hydrogel material with a stable structure.
Preferably, the molecular weight of the silk fibroin in the step (1) is selected from 5000-14000 Da, and the gel strength of the gelatin is selected from 150-300 Bloom.
Preferably, the concentration of gelatin in the fibroin/gelatin blending solution in the step (1) is 10% -50%, and the requirement of extrusion type 3D printing is met when the temperature of the liquid storage pipe in the step (3) is lower than 30 ℃; the concentration of gelatin in the silk fibroin/gelatin blending liquid in the step (1) is 0-40%, and the temperature of the liquid storage tank in the step (3) is 37-60 ℃ so as to meet the requirements of digital optical processing type 3D printing.
Preferably, the dosage of the photoinitiator in the step (2) is 0.005-6% of the mass of the silk fibroin and the gelatin.
Preferably, the photoinitiation system in the step (2) is a redox photoinitiation system selected from any one of conjugated dyes/oxidants, low oxidation state transition metal salts/persulfates and high oxidation state salts.
Preferably, the oxidizing agent is selected from one of diphenyl iodonium hexafluorophosphate, bis (4-methylbenzene) iodonium hexafluorocarbonate, bis (4-tert-butylbenzene) iodonium hexafluorocarbonate, hydrogen peroxide, sodium peroxide and potassium peroxide; the dye is selected from one of acid red 87, acid red 94, curcumin, riboflavin, eosin Y, phthalocyanine disodium, tetraphenylporphyrin ferric chloride and acridine yellow.
Preferably, the low oxidation state transition metal salt is selected from one of tris (2,2 '-bipyridine) ruthenium (ii) bis (hexafluorophosphate), tris (2, 2' -bipyridine) ruthenium (ii) chloride, (4,4 '-di-tert-butyl-2, 2' -bipyridine) bis [ (2-pyridyl) phenyl ] iridium (III) hexafluorophosphate, (2,2 '-bipyridine) bis [2- (2, 4-difluorophenyl) pyridine ] iridium (III) hexafluorophosphate, (1, 3-diaminocyclohexane) platinum dichloride, and (2, 2' -bipyridine) platinum dichloride; the persulfate is selected from one of potassium persulfate, sodium persulfate and ammonium persulfate.
Preferably, the high oxidation state salt is selected from one of 9- (2, 6-dimethylphenyl) -10-methylacridine perchlorate, 9-mesityl-10-methylacridine perchlorate, 9- (2-biphenyl) -10-methylacridine perchlorate, 9- (2, 5-dimethylphenyl) -10-methylacridine perchlorate, 10-methyl-9-phenylacridine perchlorate, 2,4, 6-triphenylpyran tetrafluoroborate and 9-mesityl-2, 7, 10-trimethylacridine perchlorate.
Preferably, the visible light source in the step (3) is selected from one of a dominant wavelength matched with a spectrum of a corresponding initiation system, specifically, a blue light LED with a dominant wavelength range of 450-480 nm, a green light LED with a dominant wavelength range of 490-570 nm, and a xenon lamp with a filter of 380-800 nm.
Preferably, the specific steps of dissolving, dialyzing, centrifuging and concentrating the silk fibroin solution obtained in the step (1) are as follows: shearing degummed raw silk grey cloth and adding CaCl with the molar ratio of 1:2:82-CH3CH2OH-H2Dissolving in an O ternary dissolution system at a bath ratio of 1:20 at 70 ℃ for 3h, and cooling to room temperature; then transferring the solution into a cellulose dialysis bag, and dialyzing in deionized water for 3-4 days; centrifuging the dialyzed silk fibroin solution for 20min to obtain a silk fibroin solution; and (3) placing the silk fibroin solution in an electric heating forced air drying box, drying and concentrating at 40 ℃, and storing at 5 ℃ for later use.
Due to the adoption of the technical scheme, the method has the following beneficial effects:
1. the invention provides a preparation method of a silk fibroin/gelatin photoinduced co-crosslinked hydrogel material suitable for 3D printing, wherein the silk fibroin and gelatin components do not need a photosensitive modification step, so that the potential influence of a photosensitive modifier, a modified catalyst, a crosslinking agent and the like on the biocompatibility of a system in the modification process is avoided;
2. the invention provides a preparation method of a silk fibroin/gelatin photoinduced co-crosslinked hydrogel material suitable for 3D printing, wherein the rheological parameters of a silk fibroin/gelatin composite printing precursor solution can be regulated, and the printed silk fibroin/gelatin co-crosslinked hydrogel material has adjustable structural strength, good shape retention and structural stability;
3. the method for preparing the silk fibroin/gelatin photoinduced co-crosslinking hydrogel material suitable for 3D printing provided by the invention can realize efficient co-crosslinking reaction of silk fibroin and gelatin by using visible light as an initiating light source, and has no radiation hazard to loaded cells and medicines.
Detailed Description
The invention aims to provide a preparation method of photoinduced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing, which has the following principle: under the induction of visible light irradiation, the excited photoinitiator and tyrosine, phenylalanine, tryptophan and other structures in the silk fibroin or gelatin structure generate electron transfer reaction to generate corresponding active free radicals, and then the active free radicals form bond reaction to realize the co-crosslinking structure between the silk fibroin and gelatin macromolecules.
The technical solution of the present invention is further illustrated by the following specific examples:
example 1:
(1) preparation of silk fibroin solution: 20g of degummed raw silk grey cloth is cut into pieces and put into CaCl with the molar ratio of 1:2:82-CH3CH2OH-H2Dissolving in a ternary O dissolving system at a bath ratio of 1:20 at 70 deg.C for 3h, and cooling to room temperature. Then transferring the solution into a cellulose dialysis bag (8000-14000 Da), and dialyzing in deionized water for 3-4 d. And (3) centrifuging the dialyzed silk fibroin solution (25 ℃, 10000r/min) for about 20min to obtain the silk fibroin solution with the mass fraction of about 1.8%. And (3) placing the silk fibroin solution in an electric heating forced air drying box, drying and concentrating at 40 ℃ until the mass fraction is 5.0%, and storing at 5 ℃ for later use.
(2) Preparing silk fibroin/gelatin blended solution: adding 1.5g gelatin particles with gel strength of 200Bloom into 8.5g silk fibroin solution, swelling for 15min, placing in a constant temperature water bath kettle, stirring at 60 deg.C, and dissolving to obtain silk fibroin/gelatin blended solution with gelatin content of 15%.
(3) Preparing a photo-induced silk fibroin/gelatin precursor: 0.0012g of tris (2, 2' -bipyridyl) ruthenium (II) chloride hexahydrate and 0.0096g of potassium persulfate are added into 10g of silk fibroin/gelatin blended liquid and stirred uniformly to prepare a photo-induced silk fibroin/gelatin precursor, and the photo-induced silk fibroin/gelatin precursor is stored in the dark for later use.
(4) Preparation of 3D printing silk fibroin/gelatin photoinduced co-crosslinked hydrogel material: and (3) placing the photo-induced silk fibroin/gelatin precursor in a liquid tank of Digital Light Processing (DLP)3D printing equipment, adjusting the temperature of the liquid tank to 39 ℃, and irradiating for 20min by using blue light of 400nm in the printing process to obtain the silk fibroin/gelatin co-crosslinked hydrogel material with a stable structure.
Example 2:
(1) preparation of silk fibroin solution: 20g of degummed raw silk grey cloth is cut into pieces and put into CaCl with the molar ratio of 1:2:82-CH3CH2OH-H2Dissolving in a ternary O dissolving system at a bath ratio of 1:20 at 70 deg.C for 3h, and cooling to room temperature. Then transferring the solution into a cellulose dialysis bag (8000-14000 Da), and dialyzing in deionized water for 3-4 d. And (3) centrifuging the dialyzed silk fibroin solution (25 ℃, 10000r/min) for about 20min to obtain the silk fibroin solution with the mass fraction of about 1.8%. And (3) placing the silk fibroin solution in an electric heating forced air drying box, drying and concentrating at 40 ℃ until the mass fraction is 8.0%, and storing at 5 ℃ for later use.
(2) Preparing silk fibroin/gelatin blended solution: adding 2.5g gelatin particles with gel strength of 230Bloom into 7.5g silk fibroin solution, swelling for 15min, placing in a constant temperature water bath kettle, stirring at 60 deg.C, and dissolving to obtain silk fibroin/gelatin blended solution with gelatin content of 25%.
(3) Preparing a photo-induced silk fibroin/gelatin precursor: and adding 0.0079g of diphenyl iodonium hexafluorophosphate and 0.0032g of riboflavin into 10g of silk fibroin/gelatin blended solution, stirring uniformly to prepare a photoinduced silk fibroin/gelatin precursor, and storing in the dark for later use.
(4) Preparation of 3D printing silk fibroin/gelatin photoinduced co-crosslinked hydrogel material: and (3) placing the photo-induced silk fibroin/gelatin precursor in a liquid tank of Digital Light Processing (DLP)3D printing equipment, adjusting the temperature of the liquid tank to 40 ℃, and irradiating for 20min by using blue light with the wavelength of 420nm in the printing process to obtain the silk fibroin/gelatin co-crosslinked hydrogel material with a stable structure.
Example 3:
(1) preparation of silk fibroin solution: 20g of degummed raw silk grey cloth is cut into pieces and put into CaCl with the molar ratio of 1:2:82-CH3CH2OH-H2Dissolving in a ternary O dissolving system at a bath ratio of 1:20 at 70 deg.C for 3h, and cooling to room temperature. Then transferring the solution into a cellulose dialysis bag (8000-14000 Da), and dialyzing in deionized water for 3-4 d. And (3) centrifuging the dialyzed silk fibroin solution (25 ℃, 10000r/min) for about 20min to obtain the silk fibroin solution with the mass fraction of about 1.8%. And (3) placing the silk fibroin solution in an electric heating forced air drying box, drying and concentrating at 40 ℃ until the mass fraction is 6.0%, and storing at 5 ℃ for later use.
(2) Preparing silk fibroin/gelatin blended solution: 1.5g of gelatin particles with the gel strength of 180Bloom are added into 8.5g of silk fibroin solution to be swelled for 15min, and then the silk fibroin/gelatin blended solution with the gelatin content of 15% is prepared by placing the silk fibroin/gelatin blended solution into a constant-temperature water bath kettle and stirring and dissolving at 60 ℃.
(3) Preparing a photo-induced silk fibroin/gelatin precursor: adding 0.0018g of 9- (2, 5-dimethylphenyl) -10-methylacridine perchlorate into 10g of silk fibroin/gelatin blended solution, stirring uniformly to prepare a light-induced silk fibroin/gelatin precursor, and storing in the dark for later use.
(4) Preparation of 3D printing silk fibroin/gelatin photoinduced co-crosslinked hydrogel material: and (3) placing the photo-induced silk fibroin/gelatin precursor in a liquid tube of an extrusion type 3D printing device, adjusting the temperature of the liquid tube to be 25 ℃, and irradiating for 20min by using 530nm green light after printing to obtain the silk fibroin/gelatin co-crosslinked hydrogel material with a stable structure.
Example 4:
(1) preparation of silk fibroin solution: 20g of degummed raw silk grey cloth is cut into pieces and put into CaCl with the molar ratio of 1:2:82-CH3CH2OH-H2Dissolving in a ternary O dissolving system at a bath ratio of 1:20 at 70 deg.C for 3h, and cooling to room temperature. Then transferring the solution to a cellulose dialysis bag (8000-14)000Da), dialyzing in deionized water for 3-4 d. And (3) centrifuging the dialyzed silk fibroin solution (25 ℃, 10000r/min) for about 20min to obtain the silk fibroin solution with the mass fraction of about 1.8%. And (3) placing the silk fibroin solution in an electric heating forced air drying box, drying and concentrating at 40 ℃ until the mass fraction is 10.0%, and storing at 5 ℃ for later use.
(2) Preparing silk fibroin/gelatin blended solution: 1.5g of gelatin particles with the gel strength of 210Bloom are added into 8.5g of silk fibroin solution to be swelled for 15min, and then the silk fibroin/gelatin blended solution with the gelatin content of 15% is prepared by placing the silk fibroin/gelatin blended solution into a constant-temperature water bath kettle and stirring and dissolving at 60 ℃.
(3) Preparing a photo-induced silk fibroin/gelatin precursor: 0.0098g of 9-mesityl-10-methylacridine perchlorate is added into 10g of silk fibroin/gelatin blended solution and stirred evenly to prepare a photo-induced silk fibroin/gelatin precursor, and the photo-induced silk fibroin/gelatin precursor is stored in the dark for later use.
(4) Preparation of 3D printing silk fibroin/gelatin photoinduced co-crosslinked hydrogel material: and (3) placing the photo-induced silk fibroin/gelatin precursor in a liquid tube of an extrusion type 3D printing device, adjusting the temperature of the liquid tube to be 20 ℃, and irradiating with 540nm green light for 20min after printing to obtain the silk fibroin/gelatin co-crosslinked hydrogel material with a stable structure.
Example 5:
(1) preparation of silk fibroin solution: 20g of degummed raw silk grey cloth is cut into pieces and put into CaCl with the molar ratio of 1:2:82-CH3CH2OH-H2Dissolving in a ternary O dissolving system at a bath ratio of 1:20 at 70 deg.C for 3h, and cooling to room temperature. Then transferring the solution into a cellulose dialysis bag (8000-14000 Da), and dialyzing in deionized water for 3-4 d. And (3) centrifuging the dialyzed silk fibroin solution (25 ℃, 10000r/min) for about 20min to obtain the silk fibroin solution with the mass fraction of about 1.8%. And (3) placing the silk fibroin solution in an electric heating forced air drying box, drying and concentrating at 40 ℃ until the mass fraction is 5.0%, and storing at 5 ℃ for later use.
(2) Preparing silk fibroin/gelatin blended solution: adding 1.5g gelatin particles with gel strength of 240Bloom into 8.5g silk fibroin solution, swelling for 15min, placing in a constant temperature water bath kettle, stirring at 60 deg.C, and dissolving to obtain silk fibroin/gelatin blended solution with gelatin content of 15%.
(3) Preparing a photo-induced silk fibroin/gelatin precursor: 0.0021g of (2, 2' -bipyridyl) bis [2- (2, 4-difluorophenyl) pyridine ] iridium (III) hexafluorophosphate hexahydrate and 0.0098g of sodium persulfate are added into 10g of silk fibroin/gelatin blended liquid and stirred uniformly to prepare a photo-induced silk fibroin/gelatin precursor which is stored in a dark place for later use.
(4) Preparation of 3D printing silk fibroin/gelatin photoinduced co-crosslinked hydrogel material: and (3) placing the photoinduced silk fibroin/gelatin precursor in a liquid tank of Digital Light Processing (DLP)3D printing equipment, adjusting the temperature of the liquid tank to be 60 ℃, and irradiating for 20min by using 400nm blue light after printing to obtain the silk fibroin/gelatin co-crosslinked hydrogel material with a stable structure.
Example 6:
(1) preparation of silk fibroin solution: 20g of degummed raw silk grey cloth is cut into pieces and put into CaCl with the molar ratio of 1:2:82-CH3CH2OH-H2Dissolving in a ternary O dissolving system at a bath ratio of 1:20 at 70 deg.C for 3h, and cooling to room temperature. Then transferring the solution into a cellulose dialysis bag (8000-14000 Da), and dialyzing in deionized water for 3-4 d. And (3) centrifuging the dialyzed silk fibroin solution (25 ℃, 10000r/min) for about 20min to obtain the silk fibroin solution with the mass fraction of about 1.8%. And (3) placing the silk fibroin solution in an electric heating forced air drying box, drying and concentrating at 40 ℃ until the mass fraction is 20.0%, and storing at 5 ℃ for later use.
(2) Preparing silk fibroin/gelatin blended solution: 1.5g of gelatin particles with the gel strength of 260Bloom are added into 8.5g of silk fibroin solution to be swelled for 15min, and then the silk fibroin/gelatin blended solution with the gelatin content of 15% is prepared by placing the silk fibroin/gelatin blended solution into a constant-temperature water bath kettle and stirring and dissolving at 60 ℃.
(3) Preparing a photo-induced silk fibroin/gelatin precursor: adding 0.0016g curcumin and 0.0098g hydrogen peroxide into 10g silk fibroin/gelatin blended solution, stirring, and making into light-induced silk fibroin/gelatin precursor, and storing in dark place for use.
(4) Preparation of 3D printing silk fibroin/gelatin photoinduced co-crosslinked hydrogel material: and (3) placing the photo-induced silk fibroin/gelatin precursor in a liquid pipe of extrusion type 3D printing equipment, adjusting the temperature of the liquid pipe to be 25 ℃, and irradiating for 20min by using 400nm blue light after printing to obtain the silk fibroin/gelatin co-crosslinked hydrogel material with a stable structure.
The above is only a specific embodiment of the present invention, but the technical features of the present invention are not limited thereto. Any simple changes, equivalent substitutions or modifications made on the basis of the present invention to solve the same technical problems and achieve the same technical effects are all covered in the protection scope of the present invention.
Claims (10)
1. A preparation method of photo-induced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing is characterized by comprising the following steps:
(1) preparation of silk fibroin/gelatin blended liquid
Dissolving, dialyzing, centrifuging and concentrating degummed raw silk grey cloth to obtain a silk fibroin solution, adding solid gelatin particles with certain gel strength and specific proportion into the silk fibroin solution with certain molecular weight for swelling for 15-30 min, and stirring at 60 ℃ until the gelatin particles are completely dissolved to obtain a silk fibroin/gelatin blended solution;
(2) preparation of photo-crosslinking silk fibroin/gelatin precursor solution
Adding a certain amount of dissolved photoinitiation system into the silk fibroin/gelatin blended solution prepared in the step (1) to prepare a photocrosslinking silk fibroin/gelatin precursor solution, and storing the solution in the dark for later use;
(3) preparation of 3D printing silk fibroin/gelatin photoinduced co-crosslinked hydrogel material
And (3) placing the photo-crosslinked silk fibroin/gelatin precursor liquid obtained in the step (2) in a liquid storage tank/pipe of a 3D printing device, adjusting the rheological property of the silk fibroin/gelatin blended liquid through temperature, then printing by using the 3D printing device to obtain a preset structure, and irradiating and crosslinking through visible light with certain intensity in the printing process/after printing to obtain the silk fibroin/gelatin co-crosslinked hydrogel material with a stable structure.
2. The method for preparing photo-induced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing according to claim 1, wherein: the molecular weight of the silk fibroin in the step (1) is selected from 5000-14000 Da, and the gel strength of the gelatin is selected from 150-300 Bloom.
3. The method for preparing photo-induced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing according to claim 1, wherein: the concentration of gelatin in the fibroin/gelatin blending solution in the step (1) is 10% -50%, and the requirement of extrusion type 3D printing is met when the temperature of the liquid storage pipe in the step (3) is lower than 30 ℃; the concentration of gelatin in the silk fibroin/gelatin blending liquid in the step (1) is 0-40%, and the temperature of the liquid storage tank in the step (3) is 37-60 ℃ so as to meet the requirements of digital optical processing type 3D printing.
4. The method for preparing photo-induced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing according to claim 1, wherein: the dosage of the photoinitiator in the step (2) is 0.005-6% of the mass of the silk fibroin and the gelatin.
5. The method for preparing photo-induced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing according to claim 1, wherein: the photoinitiation system in the step (2) is an oxidation-reduction photoinitiation system, and is selected from any one of conjugated dyes/oxidants, low oxidation state transition metal salts/persulfates and high oxidation state salts.
6. The method for preparing photo-induced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing according to claim 5, wherein: the oxidant is selected from one of diphenyl iodonium hexafluorophosphate, bis (4-methylbenzene) iodonium hexafluorophosphate, bis (4-tert-butylbenzene) iodonium hexafluorophosphate, hydrogen peroxide, sodium peroxide and potassium peroxide; the dye is selected from one of acid red 87, acid red 94, curcumin, riboflavin, eosin Y, phthalocyanine disodium, tetraphenylporphyrin ferric chloride and acridine yellow.
7. The method for preparing photo-induced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing according to claim 5, wherein: the low oxidation state transition metal salt is selected from one of tris (2,2 '-bipyridyl) ruthenium (II) bis (hexafluorophosphate), tris (2, 2' -bipyridyl) ruthenium (II) chloride, (4,4 '-di-tert-butyl-2, 2' -bipyridyl) bis [ (2-pyridyl) phenyl ] iridium (III) hexafluorophosphate, (2,2 '-bipyridyl) bis [2- (2, 4-difluorophenyl) pyridine ] iridium (III) hexafluorophosphate, (1, 3-diaminocyclohexane) platinum dichloride and (2, 2' -bipyridyl) platinum dichloride; the persulfate is selected from one of potassium persulfate, sodium persulfate and ammonium persulfate.
8. The method for preparing photo-induced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing according to claim 5, wherein: the high oxidation state salt is selected from one of 9- (2, 6-dimethylphenyl) -10-methylacridine perchlorate, 9-mesityl-10-methylacridine perchlorate, 9- (2-biphenyl) -10-methylacridine perchlorate, 9- (2, 5-dimethylphenyl) -10-methylacridine perchlorate, 10-methyl-9-phenylacridine perchlorate, 2,4, 6-triphenylpyran tetrafluoroborate and 9-mesityl-2, 7, 10-trimethylacridine perchlorate.
9. The method for preparing photo-induced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing according to claim 1 or 5, wherein: the visible light source in the step (3) is selected from one of a dominant wavelength matched with a spectrum of a corresponding initiation system, specifically a blue light LED with a dominant wavelength range of 450-480 nm, a green light LED with a dominant wavelength range of 490-570 nm and a xenon lamp with a light filter of 380-800 nm.
10. The method for preparing photo-induced silk fibroin/gelatin co-crosslinked hydrogel suitable for 3D printing according to claim 1 or 2, wherein: the specific steps of dissolving, dialyzing, centrifuging and concentrating the silk fibroin solution obtained in the step (1) are as follows: shearing degummed raw silk grey cloth and adding CaCl with the molar ratio of 1:2:82-CH3CH2OH-H2Dissolving in an O ternary dissolution system at a bath ratio of 1:20 at 70 ℃ for 3h, and cooling to room temperature; then transferring the solution into a cellulose dialysis bag, and dialyzing in deionized water for 3-4 days; centrifuging the dialyzed silk fibroin solution for 20min to obtain a silk fibroin solution; and (3) placing the silk fibroin solution in an electric heating forced air drying box, drying and concentrating at 40 ℃, and storing at 5 ℃ for later use.
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| CN114539564B (en) * | 2022-03-30 | 2023-09-12 | 武汉纺织大学 | Preparation method of rapid-gel silk fibroin-based strong hydrogel |
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| CN115260531B (en) * | 2022-08-04 | 2024-03-22 | 重庆科技学院 | Preparation method of self-curling double-layer hydrogel sheet |
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| CN115368744A (en) * | 2022-09-13 | 2022-11-22 | 大连理工大学 | Preparation and application of fibroin and structural protein polymer composite nanoparticles |
| CN115368744B (en) * | 2022-09-13 | 2023-09-12 | 大连理工大学 | Preparation and application of fibroin and structural protein polymer composite nano-particles |
| CN116655945A (en) * | 2023-02-03 | 2023-08-29 | 媄典(北京)医疗器械有限公司 | Photo-curing silk fibroin hydrogel and preparation method thereof |
| CN117327300A (en) * | 2023-08-29 | 2024-01-02 | 江苏艾玮得生物科技有限公司 | Hydrogel and preparation method thereof, organoid and culture method thereof, and drug detection method |
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