CN112126027A - Hydrogel material and preparation method and application thereof - Google Patents
Hydrogel material and preparation method and application thereof Download PDFInfo
- Publication number
- CN112126027A CN112126027A CN202011014747.4A CN202011014747A CN112126027A CN 112126027 A CN112126027 A CN 112126027A CN 202011014747 A CN202011014747 A CN 202011014747A CN 112126027 A CN112126027 A CN 112126027A
- Authority
- CN
- China
- Prior art keywords
- collagen
- hydrogel material
- solution
- solvent
- acrylic anhydride
- 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.)
- Granted
Links
- 239000000017 hydrogel Substances 0.000 title claims abstract description 44
- 239000000463 material Substances 0.000 title claims abstract description 39
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- 102000008186 Collagen Human genes 0.000 claims abstract description 60
- 108010035532 Collagen Proteins 0.000 claims abstract description 60
- 229920001436 collagen Polymers 0.000 claims abstract description 60
- 239000002202 Polyethylene glycol Substances 0.000 claims abstract description 27
- 229920001223 polyethylene glycol Polymers 0.000 claims abstract description 27
- ARJOQCYCJMAIFR-UHFFFAOYSA-N prop-2-enoyl prop-2-enoate Chemical compound C=CC(=O)OC(=O)C=C ARJOQCYCJMAIFR-UHFFFAOYSA-N 0.000 claims abstract description 25
- 125000004386 diacrylate group Chemical group 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 11
- 239000002994 raw material Substances 0.000 claims abstract description 5
- 239000000243 solution Substances 0.000 claims description 47
- 239000002904 solvent Substances 0.000 claims description 19
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 16
- 102000000503 Collagen Type II Human genes 0.000 claims description 12
- 108010041390 Collagen Type II Proteins 0.000 claims description 12
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 12
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 10
- 239000011259 mixed solution Substances 0.000 claims description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 7
- BWHMMNNQKKPAPP-UHFFFAOYSA-L potassium carbonate Chemical compound [K+].[K+].[O-]C([O-])=O BWHMMNNQKKPAPP-UHFFFAOYSA-L 0.000 claims description 6
- 238000010146 3D printing Methods 0.000 claims description 5
- 239000000203 mixture Substances 0.000 claims description 5
- NIXOWILDQLNWCW-UHFFFAOYSA-N acrylic acid group Chemical group C(C=C)(=O)O NIXOWILDQLNWCW-UHFFFAOYSA-N 0.000 claims description 4
- HFBMWMNUJJDEQZ-UHFFFAOYSA-N acryloyl chloride Chemical compound ClC(=O)C=C HFBMWMNUJJDEQZ-UHFFFAOYSA-N 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 4
- 229910000027 potassium carbonate Inorganic materials 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 3
- 238000004108 freeze drying Methods 0.000 claims description 3
- 238000002390 rotary evaporation Methods 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 230000004048 modification Effects 0.000 claims description 2
- 238000012986 modification Methods 0.000 claims description 2
- 238000001556 precipitation Methods 0.000 claims description 2
- 238000004132 cross linking Methods 0.000 abstract description 8
- 230000004071 biological effect Effects 0.000 abstract description 5
- 230000007547 defect Effects 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- 230000001939 inductive effect Effects 0.000 abstract 1
- 238000002835 absorbance Methods 0.000 description 7
- 238000004458 analytical method Methods 0.000 description 7
- 239000012620 biological material Substances 0.000 description 7
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 229920000671 polyethylene glycol diacrylate Polymers 0.000 description 6
- NHJVRSWLHSJWIN-UHFFFAOYSA-N 2,4,6-trinitrobenzenesulfonic acid Chemical compound OS(=O)(=O)C1=C([N+]([O-])=O)C=C([N+]([O-])=O)C=C1[N+]([O-])=O NHJVRSWLHSJWIN-UHFFFAOYSA-N 0.000 description 5
- 210000001519 tissue Anatomy 0.000 description 5
- 230000004663 cell proliferation Effects 0.000 description 4
- 238000010586 diagram Methods 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- 239000012224 working solution Substances 0.000 description 4
- 239000004471 Glycine Substances 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000010382 chemical cross-linking Methods 0.000 description 3
- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 description 2
- 241001529936 Murinae Species 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-MICDWDOJSA-N Trichloro(2H)methane Chemical compound [2H]C(Cl)(Cl)Cl HEDRZPFGACZZDS-MICDWDOJSA-N 0.000 description 2
- 210000000988 bone and bone Anatomy 0.000 description 2
- 210000000845 cartilage Anatomy 0.000 description 2
- 238000010609 cell counting kit-8 assay Methods 0.000 description 2
- 230000003833 cell viability Effects 0.000 description 2
- 238000012512 characterization method Methods 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 239000003431 cross linking reagent Substances 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 231100000135 cytotoxicity Toxicity 0.000 description 2
- 230000003013 cytotoxicity Effects 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 210000002901 mesenchymal stem cell Anatomy 0.000 description 2
- 210000005036 nerve Anatomy 0.000 description 2
- IOLCXVTUBQKXJR-UHFFFAOYSA-M potassium bromide Chemical compound [K+].[Br-] IOLCXVTUBQKXJR-UHFFFAOYSA-M 0.000 description 2
- 230000001376 precipitating effect Effects 0.000 description 2
- 230000008929 regeneration Effects 0.000 description 2
- 238000011069 regeneration method Methods 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- CYFSUYQRQPGQOC-UHFFFAOYSA-N trilithium (2,4,6-trimethylphenyl)-[trioxido(phenyl)-lambda5-phosphanyl]methanone Chemical compound [Li+].[Li+].[Li+].CC1=CC(=C(C(=C1)C)C(=O)P(C2=CC=CC=C2)([O-])([O-])[O-])C CYFSUYQRQPGQOC-UHFFFAOYSA-N 0.000 description 2
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 1
- 238000011481 absorbance measurement Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- OFHCOWSQAMBJIW-AVJTYSNKSA-N alfacalcidol Chemical group C1(/[C@@H]2CC[C@@H]([C@]2(CCC1)C)[C@H](C)CCCC(C)C)=C\C=C1\C[C@@H](O)C[C@H](O)C1=C OFHCOWSQAMBJIW-AVJTYSNKSA-N 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 210000001188 articular cartilage Anatomy 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000008366 buffered solution Substances 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 239000000385 dialysis solution Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 238000010899 nucleation Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000000377 silicon dioxide Substances 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F299/00—Macromolecular compounds obtained by interreacting polymers involving only carbon-to-carbon unsaturated bond reactions, in the absence of non-macromolecular monomers
-
- 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
- A61L27/18—Macromolecular materials obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds
-
- 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
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/24—Collagen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- 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
-
- 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
- A61L27/60—Materials for use in artificial skin
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F299/00—Macromolecular compounds obtained by interreacting polymers involving only carbon-to-carbon unsaturated bond reactions, in the absence of non-macromolecular monomers
- C08F299/02—Macromolecular compounds obtained by interreacting polymers involving only carbon-to-carbon unsaturated bond reactions, in the absence of non-macromolecular monomers from unsaturated polycondensates
- C08F299/026—Macromolecular compounds obtained by interreacting polymers involving only carbon-to-carbon unsaturated bond reactions, in the absence of non-macromolecular monomers from unsaturated polycondensates from the reaction products of polyepoxides and unsaturated monocarboxylic acids, their anhydrides, halogenides or esters with low molecular weight
- C08F299/028—Macromolecular compounds obtained by interreacting polymers involving only carbon-to-carbon unsaturated bond reactions, in the absence of non-macromolecular monomers from unsaturated polycondensates from the reaction products of polyepoxides and unsaturated monocarboxylic acids, their anhydrides, halogenides or esters with low molecular weight photopolymerisable compositions
-
- 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
- A61L2430/00—Materials or treatment for tissue regeneration
- A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants
-
- 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
-
- 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
- A61L2430/32—Materials or treatment for tissue regeneration for nerve reconstruction
-
- 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
- C08J2355/00—Characterised by the use of homopolymers or copolymers, obtained by polymerisation reactions only involving carbon-to-carbon unsaturated bonds, not provided for in groups C08J2323/00 - C08J2353/00
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Dermatology (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Transplantation (AREA)
- Epidemiology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Polymers & Plastics (AREA)
- Dispersion Chemistry (AREA)
- Biophysics (AREA)
- Medicinal Preparation (AREA)
- Materials For Medical Uses (AREA)
Abstract
The invention discloses a hydrogel material and a preparation method and application thereof. Wherein, the two preparation raw materials of acrylic anhydride modified collagen and polyethylene glycol diacrylate are combined and matched to realize performance complementation; specifically, the hydrogel material can be endowed with biological activity by inducing acrylic anhydride to modify collagen in the polyethylene glycol diacrylate, and the existence of the polyethylene glycol diacrylate can overcome the defect of insufficient mechanical property when the collagen is used alone; and the modified collagen can be formed by ultraviolet crosslinking and curing, the forming process is simple, the three-dimensional structure is not limited, and the application range is wide. Through the mode, the hydrogel material disclosed by the invention has excellent mechanical properties and biological activity.
Description
Technical Field
The invention relates to the technical field of biological materials, in particular to a hydrogel material and a preparation method and application thereof.
Background
Biomaterial science, as a rapidly developing interdisciplinary discipline, puts urgent demands on the performance and manufacturing processes of various biomaterials. Tissue engineering requires biomaterials with sufficient and appropriate mechanical properties to meet various applications, such as hard and soft tissue replacement. Meanwhile, the biological material also has good biocompatibility. Chemical crosslinking, as opposed to physical crosslinking, can produce sufficient structural stability to meet the various properties required for biomedical applications, where photocrosslinking has gained widespread use due to a number of advantages.
The natural biological material has the characteristics of rich source, excellent biocompatibility, good degradation performance and the like, and is widely applied to the field of tissue engineering. However, the requirements of tissue engineering on material properties are high, so that the available natural biomaterials are very limited. In a limited selection, collagen, which is a major component of the extracellular matrix, has specific functional peptide sequences that bind to receptors on the plasma membrane to direct cell differentiation and other behaviors, and is an ideal material for constructing the cell growth microenvironment. However, the use of collagen currently has two problems, namely that the crosslinking is dependent on toxic or expensive chemical crosslinking agents; and secondly, the mechanical property of the single-component collagen system is insufficient after crosslinking, and different use requirements are difficult to meet.
Disclosure of Invention
The present invention is directed to solving at least one of the problems of the prior art. Therefore, the invention provides a hydrogel material, and a preparation method and application thereof.
The technical scheme adopted by the invention is as follows:
according to the first aspect of the invention, the hydrogel material is provided, and the preparation raw materials comprise, by mass, 0.1-1% of acrylic anhydride modified collagen, 5-40% of polyethylene glycol diacrylate, 0.03-0.3% of a photoinitiator, and the balance of a solvent.
Wherein the acrylic anhydride modified collagen is acrylic anhydride modified collagen, and the acrylic anhydride modification degree (measured by a TNBS method) of the modified collagen is generally 0.08-0.38 mmol/g. And polyethylene glycol diacrylate with the molecular weight of 0.2-20 k is generally adopted.
According to some embodiments of the invention, the acrylic anhydride modified collagen is acrylic anhydride modified type II collagen.
According to some embodiments of the invention, the acrylic anhydride-modified collagen is prepared by a preparation method comprising the steps of: dissolving collagen in a first solvent to obtain a first collagen solution; and then adjusting the pH value of the first collagen solution to 7.0-8.0, adding acrylic anhydride, uniformly mixing, fully reacting, dialyzing, and freeze-drying.
According to some embodiments of the invention, the first solvent is an aqueous hydrochloric acid solution or an aqueous acetic acid solution.
According to some embodiments of the present invention, the polyethylene glycol diacrylate is prepared by a preparation method comprising the steps of: dissolving polyethylene glycol and potassium carbonate in a second solvent to obtain a mixed solution; dissolving acryloyl chloride in a second solvent, and then dropwise adding the solution into the mixed solution for reaction; filtering and taking supernatant after the reaction is finished, and removing most of the second solvent by rotary evaporation; then ether precipitation and washing are adopted, and then drying treatment is carried out.
According to some embodiments of the invention, the second solvent is anhydrous dichloromethane. As the photoinitiator, lithium phenyl-2, 4, 6-trimethylbenzoylphosphite (LAP) can be used.
According to some embodiments of the invention, the solvent is aqueous hydrochloric acid.
In a second aspect of the present invention, there is provided a method for preparing any one of the hydrogel materials provided in the first aspect of the present invention, comprising the steps of:
s1, dissolving the acrylic anhydride modified collagen in a solvent, adding a photoinitiator, and uniformly mixing to obtain a collagen solution;
s2, adding polyethylene glycol diacrylate into the collagen solution, and stirring and dissolving the mixture in a dark place to obtain a mixed solution;
and S3, carrying out photocrosslinking reaction on the mixed solution under ultraviolet light.
In a third aspect of the invention, there is provided a use of any one of the hydrogel materials provided in the first aspect of the invention in 3D printing.
The embodiment of the invention has the beneficial effects that:
the embodiment of the invention provides a hydrogel material, which can realize performance complementation by combining and matching two preparation raw materials of acrylic anhydride modified collagen and polyethylene glycol diacrylate. Specifically, the hydrogel material can be endowed with biological activity by introducing acrylic anhydride modified collagen into polyethylene glycol diacrylate, and the existence of the polyethylene glycol diacrylate can solve the problem of insufficient mechanical property when the collagen is used alone; in addition, the collagen modified by the acrylic anhydride (namely the collagen modified by the acrylic anhydride) can overcome the defect that the crosslinking of the collagen depends on toxic or expensive chemical crosslinking agents, and the modified collagen can be crosslinked and cured by ultraviolet light, so that the forming process is simple, the three-dimensional structure is not limited, and the application range is wide. Through the mode, the hydrogel material has excellent mechanical property and biological activity, and is expected to be used as a tissue engineering material for regeneration of bones, cartilages, skins, nerves and the like.
Drawings
FIG. 1 is a schematic diagram illustrating the synthesis of a hydrogel material according to one embodiment of the present invention;
FIG. 2 is a schematic diagram of electrophoresis of type II collagen used in the preparation of hydrogel materials in accordance with one embodiment of the present invention;
FIG. 3 shows the polyethylene glycol diacrylate prepared in examples 6 to 8 of the present invention1H nuclear magnetic analysis chart;
FIG. 4 is a graph showing the infrared analysis of the polyethylene glycol diacrylate prepared in examples 6 to 8 of the present invention;
FIG. 5 is a graph showing photo-crosslinking performance analysis of a collagen solution in example 9 of the present invention;
FIG. 6 is a graph showing shear modulus analysis of hydrogel materials according to examples 9 to 12 of the present invention;
FIG. 7 is a graph showing cytotoxicity analysis of hydrogel materials according to examples 9 to 12 of the present invention;
FIG. 8 is a diagram showing cell proliferation analysis of hydrogel materials according to examples 9 to 12 of the present invention.
Detailed Description
The concept and technical effects of the present invention will be clearly and completely described below in conjunction with the embodiments to fully understand the objects, features and effects of the present invention. It is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments, and those skilled in the art can obtain other embodiments without inventive effort based on the embodiments of the present invention, and all embodiments are within the protection scope of the present invention.
The invention provides a preparation method of a hydrogel material, a schematic preparation flow diagram of which is shown in figure 1, and the preparation method specifically comprises the following steps:
preparation of (I) acrylic anhydride modified type II collagen
The method is characterized in that II type collagen extracted from pig articular cartilage is taken as a raw material, the extracted II type collagen is subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the purity and the molecular weight of the extracted II type collagen are characterized, and the obtained result is shown in figure 2.
As can be seen from FIG. 2, the obtained type II collagen has beta chain and gamma chain, only one alpha chain, and no staining band below 100kDa, which accords with the characteristics of type II collagen, and the purity of the separated type II collagen is high.
The specific method for modifying the type II collagen by the acrylic anhydride comprises the following steps:
s1, dissolving 0.25g type II Collagen (Collagen) in 100mL of 10mM HCl solution, and stirring at 4 ℃ until the Collagen is dissolved to obtain a first Collagen solution;
s2, adding 0.2M Na into the first collagen solution in a divided manner and slowly2HPO4Adjusting the pH value of the solution to 7.5;
s3, adding acrylic anhydride into the first collagen solution processed in the step S2 according to the addition amount shown in the table 1, uniformly stirring, and reacting for 8 hours;
s4, after the reaction is finished, the reaction solution is filled into a dialysis bag and dialyzed in 10mM HCl solution at 4 ℃ for three days, and the dialysis solution is replaced every 12 h.
And S5, and freeze-drying the obtained solution after dialysis to obtain white sponge-like solid, namely the acrylic anhydride modified type II Collagen (Collagen-AA).
The degree of substitution of the amino groups of collagen was measured by TNBS (2,4,6-Trinitrobenzenesulfonic acid) method for the acrylic anhydride-modified type II collagen synthesized as described above.
The TNBS method is specifically performed as follows;
1. solution to be tested: several samples to be tested and standard samples were dissolved in 10mM hydrochloric acid solution, 0.1M borate buffer solution with pH 8 was added, and 1M sodium hydroxide was used to adjust pH 8 to prepare a solution to be tested of 2.5 mg/mL.
2. Working solution: accurately weigh 60mg glycine and formulate 100mL of 8mM glycine solution with 0.1M borate buffered solution at pH 8; then, 0, 0.5mL, 1mL, 1.5mL, and 2mL of the solution were diluted to 10mL and prepared into glycine solutions with concentrations of 0, 0.4mM, 0.8mM, 1.2mM, and 1.6mM, respectively, for use.
3. And (3) absorbance measurement: respectively adding 1mL of solution to be detected or working solution and 1mL of 1% TNBS solution into each centrifuge tube with a plug, and reacting for 2h at 37 ℃; after the reaction is finished, adding 0.5mL of 1M hydrochloric acid solution into each centrifuge tube to stop the reaction, and adding 1mL of 10% sodium dodecyl sulfate solution to prevent the solution from precipitating; taking a plurality of solutions to be detected or working solutions in each centrifugal tube, and measuring absorbance at 345nm by using an enzyme-labeling instrument; and drawing a working curve of the amino content and the absorbance of the working solution, and calculating the amino content in the sample to be detected according to the absorbance of the solution to be detected, thereby indirectly calculating the amino substitution degree, wherein the obtained result is shown in table 1.
TABLE 1
Preparation of (di) polyethylene glycol diacrylate
The specific method comprises the following steps:
s1, drying polyethylene glycol (PEG) at 50 ℃ in vacuum overnight, K2CO3Grinding, vacuum drying at 100 deg.C overnight, and cooling in vacuum;
s2, 50g of PEG was dissolved in 100mL of anhydrous dichloromethane in a 500mL three-necked flask, and K was added in the amount shown in Table 22CO3Fully stirring and dispersing;
s3, dissolving acryloyl chloride in anhydrous dichloromethane (1:10v/v) according to the addition amount shown in the table 2, dropwise and slowly adding the solution into a three-neck flask, and reacting at room temperature for 24 hours under the protection of nitrogen;
s4, filtering the mixture to remove solids, performing rotary evaporation on the supernatant at 37 ℃ to remove most of the solvent, fully precipitating with diethyl ether, washing with diethyl ether for 3-5 times, and finally completely drying in vacuum to obtain the product, namely the polyethylene glycol diacrylate (PEGDA).
TABLE 2
Sample (I) | Molecular weight of PEG | K2CO3Mass/g | Acryloyl chloride volume/mL |
Example 6 | 1.5k | 13.8 | 8.1 |
Example 7 | 4k | 5.2 | 3.0 |
Example 8 | 10k | 2.1 | 1.2 |
Dissolving the polyethylene glycol diacrylate prepared in examples 6 to 8 in deuterated chloroform1The H NMR spectrum was characterized, and the obtained results are shown in FIG. 3. As can be seen from fig. 3, the chemical shifts can be well assigned to the corresponding protons, ═ 6.5, 6.2, 5.9(6H, H)2C ═ CH-), 3.6(PEG chain protons).
In addition, a certain amount of the polyethylene glycol diacrylate prepared in the above examples 6 to 8 and potassium bromide ground tablets were subjected to Fourier infrared analysis and characterization, and the obtained results are shown in FIG. 4. The infrared absorption peak is 2883cm as can be seen from FIG. 4-1(-CH2-),1724cm-1(-C=O),1635cm-1(H2C=CH-)。
(III) preparation of hydrogel Material
The preparation method comprises the following steps:
s1, adopting the acrylic anhydride modified II type collagen prepared in the embodiment 3, and violently stirring the modified II type collagen at 4 ℃ and dissolving the modified II type collagen in 20mM HCl to prepare a collagen solution with the mass concentration of 1%;
s2, 1mL of 1% collagen solution is added with 20mM HCl and 0.3% LAP solution accounting for 1/10 of the volume of the final solution to be diluted into collagen solutions with different concentrations (shown in Table 3);
s3, adding 200mg of the 1.5k polyethylene glycol diacrylate with the molecular weight prepared in the example 6 into the prepared collagen solution, and stirring and dissolving the mixture at 4 ℃ in dark to ensure that the mass concentration of the mixture is 10 percent;
s4, transferring the uniform transparent bubble-free mixed solution to silica gelPlacing in a mold, and placing in ultraviolet light (365nm, 4800 μ w/cm)2) Irradiating for 10min, and taking out the cross-linked slice after cooling to room temperature;
s5, soaking the hydrogel obtained by crosslinking in a PBS solution for two days, and changing the PBS one day to obtain the colorless transparent neutral hydrogel (PEGDA/Col-AA hydrogel).
TABLE 3
The photo-crosslinking performance characterization of the collagen solution with the above concentration of 0.25% was performed, and the obtained results are shown in fig. 5, where G' is the storage modulus and G ″ is the loss modulus. As can be seen from fig. 5, the collagen solution at this concentration can be crosslinked in response to ultraviolet light, and it can be understood that the acrylic anhydride-modified type II collagen at higher concentration in examples 10 to 16 can also be crosslinked in response to ultraviolet light.
The hydrogel materials prepared in examples 9 to 12 were prepared into disks with a diameter of 20mm and a thickness of 1mm, and the shear modulus was measured at 37 ℃ and γ of 1% at 0.1 to 100rad/s on a rotational rheometer, specifically, the storage modulus G' and the loss modulus G ″ were measured as a function curve of frequency, and the results are shown in fig. 6, where PEGDA indicates no collagen was added in fig. 6. As can be seen from fig. 6, the shear modulus of the hydrogel significantly increased as the content of the collagen component increased.
The hydrogel materials prepared in examples 9-12 were prepared into disks with a diameter of 5mm and a thickness of 1mm for cytotoxicity experiments. Specifically, by culturing murine mesenchymal stem cells in a 24-well plate at a density of about 20000 cells per square centimeter, the hydrogel was soaked in 1mL of the medium by trans-well without contact with the cells. The same density of cell seeding wells without hydrogel served as positive controls. CCK-8 assays were performed on days 1, 4 and 7 after inoculation. Absorbance at 450nm was measured by a microplate reader. Cell viability was calculated using the following formula: cell viability (%) ═ (OD)Test specimen/ODControl sample) X 100%. The results are shown in FIG. 7, in which the 0% group represents the PEGDA group to which no collagen was added. From FIG. 7As can be seen, none of the hydrogel materials prepared in examples 9-12 above were cytotoxic.
Cell proliferation experiments were performed on the hydrogel materials obtained in examples 9 to 12, and specifically, murine bone marrow mesenchymal stem cells were seeded on the hydrogel surface in a 48-well plate at a density of about 20000 cells per square centimeter. CCK-8 assays were performed at 4h and 1, 4 and 7 days after inoculation. Specifically, absorbance at 450nm was measured using a microplate reader, and the cell proliferation rate was calculated using the following formula: cell proliferation rate (%) ═ (OD)sample/ODoriginal) X 100% where ODoriginalIs the absorbance, OD, corresponding to 4h of culture on the surface of the hydrogelsampleThe absorbance of the cells was measured for 1, 4 and 7 days after culturing on the hydrogel surface. The results are shown in FIG. 8, in which the 0% group represents the PEGDA group without collagen addition. As can be seen from fig. 8, the cells proliferated more in the collagen-containing group at 4 days and 7 days, compared to the collagen-free PEGDA group.
From the above, the hydrogel material prepared by the above method of the present invention has excellent mechanical properties and biological activity, and is expected to be used as a tissue engineering material for bone, cartilage, skin, nerve regeneration, etc.; and can be used as 3D printing ink or used for preparing 3D printing ink, and further used for 3D printing.
Claims (10)
1. The hydrogel material is characterized by comprising the following preparation raw materials in percentage by mass: 0.1-1% of acrylic anhydride modified collagen, 5-40% of polyethylene glycol diacrylate, 0.03-0.3% of photoinitiator and the balance of solvent.
2. The hydrogel material of claim 1, wherein the acrylic anhydride modified collagen is acrylic anhydride modified type II collagen.
3. The hydrogel material according to claim 1, wherein the modification degree of the acrylic anhydride-modified collagen is 0.08 to 0.38 mmol/g.
4. The hydrogel material according to claim 1, wherein the acrylic anhydride modified collagen is prepared by a preparation method comprising the steps of: dissolving collagen in a first solvent to obtain a first collagen solution; and then adjusting the pH value of the first collagen solution to 7.0-8.0, adding acrylic anhydride, uniformly mixing, fully reacting, dialyzing, and freeze-drying.
5. The hydrogel material of claim 4, wherein the first solvent is an aqueous hydrochloric acid solution or an aqueous acetic acid solution.
6. The hydrogel material of claim 1, wherein the polyethylene glycol diacrylate is prepared by a preparation method comprising the steps of: dissolving polyethylene glycol and potassium carbonate in a second solvent to obtain a mixed solution; dissolving acryloyl chloride in a second solvent, and then dropwise adding the solution into the mixed solution for reaction; filtering and taking supernatant after the reaction is finished, and removing most of the second solvent by rotary evaporation; then ether precipitation and washing are adopted, and then drying treatment is carried out.
7. The hydrogel material of claim 6, wherein the second solvent is anhydrous dichloromethane.
8. The hydrogel material according to any one of claims 1 to 7, wherein the solvent is an aqueous hydrochloric acid solution.
9. The method for preparing a hydrogel material according to any one of claims 1 to 8, comprising the steps of:
s1, dissolving the acrylic anhydride modified collagen in a solvent, adding a photoinitiator, and uniformly mixing to obtain a collagen solution;
s2, adding polyethylene glycol diacrylate into the collagen solution, and stirring and dissolving the mixture in a dark place to obtain a mixed solution;
and S3, carrying out photocrosslinking reaction on the mixed solution under ultraviolet light.
10. Use of the hydrogel material of any one of claims 1 to 8 in 3D printing.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011014747.4A CN112126027B (en) | 2020-09-24 | 2020-09-24 | Hydrogel material and preparation method and application thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011014747.4A CN112126027B (en) | 2020-09-24 | 2020-09-24 | Hydrogel material and preparation method and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN112126027A true CN112126027A (en) | 2020-12-25 |
CN112126027B CN112126027B (en) | 2022-04-08 |
Family
ID=73840163
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202011014747.4A Active CN112126027B (en) | 2020-09-24 | 2020-09-24 | Hydrogel material and preparation method and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN112126027B (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113425902A (en) * | 2021-07-19 | 2021-09-24 | 陈基施展 | Visible light crosslinked collagen biological ink capable of being printed in 3D mode and preparation method thereof |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20020192289A1 (en) * | 2001-06-18 | 2002-12-19 | Ji Zheng | Polymer gel for cancer treatment |
CN107236135A (en) * | 2017-07-07 | 2017-10-10 | 中国科学院理化技术研究所 | Gelatin hydrogel and preparation method and application thereof |
CN108003360A (en) * | 2017-10-16 | 2018-05-08 | 四川大学 | Stem cell is induced into the preparation method of the II Collagen Type VI hydrogels of cartilage differentiation |
-
2020
- 2020-09-24 CN CN202011014747.4A patent/CN112126027B/en active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20020192289A1 (en) * | 2001-06-18 | 2002-12-19 | Ji Zheng | Polymer gel for cancer treatment |
CN107236135A (en) * | 2017-07-07 | 2017-10-10 | 中国科学院理化技术研究所 | Gelatin hydrogel and preparation method and application thereof |
CN108003360A (en) * | 2017-10-16 | 2018-05-08 | 四川大学 | Stem cell is induced into the preparation method of the II Collagen Type VI hydrogels of cartilage differentiation |
Non-Patent Citations (2)
Title |
---|
YIHU WANG: "Development of a Photo-Crosslinking, Biodegradable GelMA/PEGDA Hydrogel for Guided Bone Regeneration Materials", 《MATERIALS》 * |
王荧: "载药胶原/聚乙二醇二丙烯酸酯复合水凝胶的研究", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113425902A (en) * | 2021-07-19 | 2021-09-24 | 陈基施展 | Visible light crosslinked collagen biological ink capable of being printed in 3D mode and preparation method thereof |
Also Published As
Publication number | Publication date |
---|---|
CN112126027B (en) | 2022-04-08 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Hoch et al. | Stiff gelatin hydrogels can be photo-chemically synthesized from low viscous gelatin solutions using molecularly functionalized gelatin with a high degree of methacrylation | |
Zhang et al. | Synthesis and characterization of hyaluronic acid/human-like collagen hydrogels | |
US20210001009A1 (en) | Biogum and botanical gum hydrogel bioinks for the physiological 3d bioprinting of tissue constructs for in vitro culture and transplantation | |
CN109316630B (en) | 3D printing ink of biological bionic matrix and preparation method thereof | |
CN112190763B (en) | Hyaluronic acid/epsilon-polylysine antibacterial hydrogel and preparation method and application thereof | |
CN114796604B (en) | 3D printing ink for cornea regeneration and preparation method and application thereof | |
CN111218011B (en) | Polyethylene glycol-based hydrogel and preparation method and application thereof | |
CN113444264B (en) | Preparation method and application method of double-network hydrogel for three-dimensional cell culture | |
CN112126027B (en) | Hydrogel material and preparation method and application thereof | |
CN110818921A (en) | Rapidly-curable double-crosslinked hydrogel and preparation method and application thereof | |
CN108341976A (en) | The derivative and synthetic method of methyl-prop alkylene host material based on click chemistry | |
CN107118373A (en) | A kind of POSS PEG hybridized hydrogels, its preparation method and application | |
CN106798949A (en) | A kind of porous hydroxyapatite bone renovating material and preparation method thereof | |
Tirella et al. | Functionalized enzyme-responsive biomaterials to model tissue stiffening in vitro | |
CN113583455B (en) | Collagen-modified chitosan double-network hydrogel, biological ink, preparation method and application | |
CN110713601B (en) | Quick-curing hydrogel based on bioorthogonal reaction, preparation method and application thereof | |
CN113265032B (en) | Preparation method and application of polyallylamine-modified temperature-sensitive copolymer | |
Zhao et al. | Rapid printing of 3D porous scaffolds for breast reconstruction | |
CN110746516B (en) | Natural polymer hydrogel based on collagen and preparation method thereof | |
Huang et al. | Modified mannan for 3D bioprinting: a potential novel bioink for tissue engineering | |
CN108084466B (en) | Composite membrane based on egg white and methacrylic acid derivative polymer and application of composite membrane in stem cell culture | |
CN111529755A (en) | POSS (polyhedral oligomeric silsesquioxane) reinforced hydrogel as well as preparation method and application thereof | |
CN115887772A (en) | Gelatin/sodium alginate hydrogel-based 3D printing biological ink and application thereof | |
CN115475279B (en) | Photosensitive cartilage acellular matrix hydrogel material, and preparation method and application thereof | |
CN108164713A (en) | Degradable and water soluble POSS-PEG hybridized hydrogels and its preparation method and application |
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 |