CN111748120A - 一种聚多巴胺掺杂葡聚糖水凝胶多孔支架及制备方法及应用 - Google Patents
一种聚多巴胺掺杂葡聚糖水凝胶多孔支架及制备方法及应用 Download PDFInfo
- Publication number
- CN111748120A CN111748120A CN202010487754.XA CN202010487754A CN111748120A CN 111748120 A CN111748120 A CN 111748120A CN 202010487754 A CN202010487754 A CN 202010487754A CN 111748120 A CN111748120 A CN 111748120A
- Authority
- CN
- China
- Prior art keywords
- polydopamine
- glucan
- doped
- porous scaffold
- solution
- 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.)
- Pending
Links
- 239000000017 hydrogel Substances 0.000 title claims abstract description 49
- 229920001503 Glucan Polymers 0.000 title claims abstract description 37
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims abstract description 69
- 239000000499 gel Substances 0.000 claims abstract description 26
- 229920001690 polydopamine Polymers 0.000 claims abstract description 22
- CTENFNNZBMHDDG-UHFFFAOYSA-N Dopamine hydrochloride Chemical compound Cl.NCCC1=CC=C(O)C(O)=C1 CTENFNNZBMHDDG-UHFFFAOYSA-N 0.000 claims abstract description 14
- 229960001149 dopamine hydrochloride Drugs 0.000 claims abstract description 14
- 239000003431 cross linking reagent Substances 0.000 claims abstract description 11
- CUGZWHZWSVUSBE-UHFFFAOYSA-N 2-(oxiran-2-ylmethoxy)ethanol Chemical compound OCCOCC1CO1 CUGZWHZWSVUSBE-UHFFFAOYSA-N 0.000 claims abstract description 3
- 238000002156 mixing Methods 0.000 claims abstract description 3
- 229920002307 Dextran Polymers 0.000 claims description 31
- 239000000243 solution Substances 0.000 claims description 31
- 239000007864 aqueous solution Substances 0.000 claims description 20
- 239000011148 porous material Substances 0.000 claims description 11
- 238000003756 stirring Methods 0.000 claims description 9
- 238000007254 oxidation reaction Methods 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 6
- 239000011259 mixed solution Substances 0.000 claims description 5
- 230000008439 repair process Effects 0.000 claims description 4
- 239000003102 growth factor Substances 0.000 claims description 3
- 229920000715 Mucilage Polymers 0.000 claims description 2
- 239000000853 adhesive Substances 0.000 claims description 2
- 239000003242 anti bacterial agent Substances 0.000 claims description 2
- 239000003963 antioxidant agent Substances 0.000 claims description 2
- 230000003078 antioxidant effect Effects 0.000 claims description 2
- 230000010261 cell growth Effects 0.000 claims description 2
- 239000003995 emulsifying agent Substances 0.000 claims description 2
- 239000000796 flavoring agent Substances 0.000 claims description 2
- 235000013355 food flavoring agent Nutrition 0.000 claims description 2
- 235000003599 food sweetener Nutrition 0.000 claims description 2
- 239000003906 humectant Substances 0.000 claims description 2
- 239000003755 preservative agent Substances 0.000 claims description 2
- 230000002335 preservative effect Effects 0.000 claims description 2
- 239000003765 sweetening agent Substances 0.000 claims description 2
- 239000002562 thickening agent Substances 0.000 claims description 2
- 230000037314 wound repair Effects 0.000 abstract description 5
- 238000000034 method Methods 0.000 abstract 2
- 238000000338 in vitro Methods 0.000 abstract 1
- 239000000203 mixture Substances 0.000 abstract 1
- 230000001590 oxidative effect Effects 0.000 abstract 1
- 238000006116 polymerization reaction Methods 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 20
- VYFYYTLLBUKUHU-UHFFFAOYSA-N dopamine Chemical compound NCCC1=CC=C(O)C(O)=C1 VYFYYTLLBUKUHU-UHFFFAOYSA-N 0.000 description 10
- 241000700159 Rattus Species 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 229960003638 dopamine Drugs 0.000 description 5
- 210000002950 fibroblast Anatomy 0.000 description 5
- 241000237536 Mytilus edulis Species 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 235000020638 mussel Nutrition 0.000 description 4
- WEXRUCMBJFQVBZ-UHFFFAOYSA-N pentobarbital Chemical compound CCCC(C)C1(CC)C(=O)NC(=O)NC1=O WEXRUCMBJFQVBZ-UHFFFAOYSA-N 0.000 description 4
- 206010052428 Wound Diseases 0.000 description 3
- 208000027418 Wounds and injury Diseases 0.000 description 3
- YCIMNLLNPGFGHC-UHFFFAOYSA-N catechol Chemical group OC1=CC=CC=C1O YCIMNLLNPGFGHC-UHFFFAOYSA-N 0.000 description 3
- 238000004132 cross linking Methods 0.000 description 3
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000003760 magnetic stirring Methods 0.000 description 3
- 239000002953 phosphate buffered saline Substances 0.000 description 3
- 206010002091 Anaesthesia Diseases 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 230000037005 anaesthesia Effects 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 229960001412 pentobarbital Drugs 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- HPALAKNZSZLMCH-UHFFFAOYSA-M sodium;chloride;hydrate Chemical compound O.[Na+].[Cl-] HPALAKNZSZLMCH-UHFFFAOYSA-M 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 230000029663 wound healing Effects 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102000008934 Muscle Proteins Human genes 0.000 description 1
- 108010074084 Muscle Proteins Proteins 0.000 description 1
- 206010072170 Skin wound Diseases 0.000 description 1
- 208000031737 Tissue Adhesions Diseases 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 238000001949 anaesthesia Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 229940124350 antibacterial drug Drugs 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 238000003501 co-culture Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 125000002791 glucosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)[C@H](O1)CO)* 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 210000001243 pseudopodia Anatomy 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000001878 scanning electron micrograph Methods 0.000 description 1
- 239000013535 sea water Substances 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- 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
- C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
- C08J9/0061—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof characterized by the use of several polymeric components
-
- 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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0009—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form containing macromolecular materials
- A61L26/0052—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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/0066—Medicaments; Biocides
-
- 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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/008—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
- A61L26/00—Chemical aspects of, or use of materials for, wound dressings or bandages in liquid, gel or powder form
- A61L26/0061—Use of materials characterised by their function or physical properties
- A61L26/0085—Porous materials, e.g. foams or sponges
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/0062—General methods for three-dimensional culture
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
- C12N5/0656—Adult fibroblasts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
- A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
- A61L2300/412—Tissue-regenerating or healing or proliferative agents
-
- 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
- C08J2305/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2301/00 or C08J2303/00
- C08J2305/02—Dextran; 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
- C08J2379/00—Characterised by the use of macromolecular compounds obtained by reactions forming in the main chain of the macromolecule a linkage containing nitrogen with or without oxygen, or carbon only, not provided for in groups C08J2361/00 - C08J2377/00
- C08J2379/04—Polycondensates having nitrogen-containing heterocyclic rings in the main chain; Polyhydrazides; Polyamide acids or similar polyimide precursors
-
- 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
- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/02—Dextran; 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
- C08J2479/00—Characterised by the use of macromolecular compounds obtained by reactions forming in the main chain of the macromolecule a linkage containing nitrogen with or without oxygen, or carbon only, not provided for in groups C08J2461/00 - C08J2477/00
- C08J2479/04—Polycondensates having nitrogen-containing heterocyclic rings in the main chain; Polyhydrazides; Polyamide acids or similar polyimide precursors
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2513/00—3D culture
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/30—Synthetic polymers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2533/00—Supports or coatings for cell culture, characterised by material
- C12N2533/70—Polysaccharides
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Materials Engineering (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Dispersion Chemistry (AREA)
- Cell Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Polymers & Plastics (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Rheumatology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明涉及一种聚多巴胺掺杂葡聚糖水凝胶多孔支架及制备方法及应用，用于细胞体外3D培养及伤口修复，水凝胶支架由以下方法制备得到：使用氢氧化钠溶液溶解葡聚糖，制备葡聚糖溶液，将盐酸多巴胺加入到氢氧化钠溶液氧化聚合生成聚多巴胺溶液，两者按一定比例混合均匀后加入交联剂乙二醇缩水甘油醚，置于37℃恒温箱中12小时后形成凝胶，本发明中所用的葡聚糖是一种细菌分泌的高水溶性天然多糖，具有良好的生物相容性和可降解性，形成凝胶后生成大量微孔结构，具有的大比表面积可为聚多巴胺提供大量的负载位点，有利于细胞进入至凝胶内部，与聚多巴胺产生黏附作用，是一种优异的多孔水凝胶支架材料，在伤口修复及生物组织工程上具有广阔的应用前景。
Description
技术领域
本发明涉及生物支架领域，尤其是涉及一种聚多巴胺掺杂葡聚糖水凝胶多孔支架及制备方法及应用。
背景技术
葡聚糖是某些微生物在生长过程中分泌的一种以葡萄糖为单糖组成的微生物多糖，其中葡萄糖单元之间以糖苷键连接。葡聚糖具有很好的生物相容性和可降解性，但是葡聚糖凝胶的黏附能力较弱，在实际应用中易与机体脱落，从而阻碍其进一步应用。
研究发现，海洋生物贻贝通过其足腺分泌的具有超强黏附性能的蛋白，可在海水等潮湿环境中牢固黏附在各种材料的表面，聚多巴胺是一种贻贝仿生类材料，具有与贻贝黏附蛋白相似的结构和超强的黏附性能。
水凝胶支架应用于生物医学领域，不仅需要优异的生物相容性，还需要具有一定的力学性能和生物学功能，以适用于不同的医用领域和临床需求。开发一种简单快捷、成本低廉的多功能水凝胶多孔支架的需求仍然十分迫切，也是当前水凝胶多孔支架应用于临床实践亟待解决的技术问题。
发明内容
针对现有技术存在的不足，本发明的目的在于提供一种聚多巴胺掺杂葡聚糖水凝胶多孔支架及制备方法及应用。
为实现上述目的，本发明提供一种聚多巴胺掺杂葡聚糖水凝胶多孔支架的制备方法，将盐酸多巴胺固体溶于1.2M的氢氧化钠水溶液中，在常温条件下搅拌进行氧化反应制备含30mg/mL的盐酸多巴胺氢氧化钠水溶液，将葡聚糖溶于1.2M的氢氧化钠水溶液中，在常温条件下搅拌进行氧化反应制备含15％(w/v)葡聚糖溶液，将盐酸多巴胺氢氧化钠水溶液和葡聚糖溶液混合搅拌，加入与混合液体积比为1∶10的交联剂，所述盐酸多巴胺氢氧化钠水溶液和葡聚糖溶液体积比为1-5∶5。
进一步的，所述交联剂为乙二醇缩水甘油醚。
本发明还提供上述制备方法制备的聚多巴胺掺杂葡聚糖水凝胶多孔支架。
本发明还提供上述聚多巴胺掺杂葡聚糖水凝胶多孔支架的应用，聚多巴胺掺杂葡聚糖水凝胶多孔支架用于制备细胞支架的应用。
作为本发明的一种应用方式，所述细胞支架的孔隙为5～30μm。
本发明还提供上述聚多巴胺掺杂葡聚糖水凝胶多孔支架的应用，聚多巴胺掺杂葡聚糖水凝胶多孔支架用于制备皮肤修复凝胶的应用。
作为本发明的一种应用方式，所述皮肤修复凝胶负载有细胞生长因子、抗菌剂、保湿剂、防腐剂、抗氧剂、乳化剂、增稠剂、甜味剂、胶浆剂、芳香剂以及矫味剂中的一种或多种。
本发明具有如下优点：葡聚糖具有很好的生物相容性和可降解性，成胶后生成的多孔葡聚糖凝胶能为细胞提供生长位点，且多糖材料能为细胞的生长提供一定的营养物质，是一种理想的细胞支架材料，在细胞3D培养和伤口修复中具有很好的应用前景，聚多巴胺(PDA)由于具有与贻贝的足肌蛋白相似的官能团(邻苯二酚基团)，可以显著增强材料的粘合性能，聚多巴胺的邻苯二酚基团可以通过氢键，静电相互作用和π-π堆积与各种表面相互作用，使得凝胶具有显着的组织粘附特性，将其掺杂至葡聚糖凝胶中后，有望能够促进细胞在凝胶中的黏附及防止凝胶载体在机体上的脱离，此外，聚多巴胺的掺杂能够增强葡聚糖凝胶的力学性能，调节凝胶孔径大小，结合葡聚糖与聚多巴胺的优点，有望构筑性能优异且实用性强的水凝胶多孔支架，本发明所构筑的水凝胶多孔支架在细胞3D培养和伤口修复等生物医学领域具有广阔的应用前景。
附图说明
图1为葡聚糖/聚多巴胺水凝胶支架的储能模量；
图2为葡聚糖/聚多巴胺水凝胶支架的扫描电镜图；
图3为成纤维细胞与葡聚糖/多巴胺水凝胶共培养后LIVE/DEAD染色分析结果；
图4为空白组(灭菌PBS)和水凝胶组(D4)对大鼠的伤口修复照片。
具体实施方式
下面将结合实施例和效果例对本发明做进一步的详述，而非限制本发明的范围。
实施例1：葡聚糖/聚多巴胺多孔水凝胶细胞支架的制备
其制备方法步骤如下：
1)盐酸多巴胺氢氧化钠水溶液是通过将30g盐酸多巴胺固体溶于1L的1.2M氢氧化钠水溶液中，在空气条件下(25℃，磁力搅拌24h)进行氧化反应来制备；
2)使用氢氧化钠溶液(1.2M)溶解葡聚糖，制备浓度为15％(w/v)的葡聚糖溶液；
3)取2mL步骤1中的盐酸多巴胺氢氧化钠水溶液和5mL步骤2中葡聚糖溶液以及3mL的1.2M的氢氧化钠溶液混合在一起，室温搅拌均匀。
4)向步骤(3)中所制备的混合溶液中加入1.0mL交联剂EGDE，搅拌均匀后置于37℃恒温箱中，交联12h形成凝胶，记为D2。
实施例2：葡聚糖/聚多巴胺多孔水凝胶细胞支架的制备
其制备方法步骤如下：
1)盐酸多巴胺氢氧化钠水溶液是通过将30g盐酸多巴胺固体溶于1L的1.2M氢氧化钠水溶液中，在空气条件下(25℃，磁力搅拌24h)进行氧化反应来制备；
2)使用氢氧化钠溶液(1.2M)溶解葡聚糖，制备浓度为15％(w/v)的葡聚糖溶液；
3)取3mL步骤1中的盐酸多巴胺氢氧化钠水溶液和5mL步骤2中葡聚糖溶液以及2mL的1.2M的氢氧化钠溶液混合在一起，室温搅拌均匀。
4)向步骤(3)中所制备的混合溶液中加入1.0mL交联剂EGDE，搅拌均匀后置于37℃恒温箱中，交联12h形成凝胶，记为D3。
实施例3：葡聚糖/聚多巴胺多孔水凝胶细胞支架的制备
其制备方法步骤如下：
1)盐酸多巴胺氢氧化钠水溶液是通过将30g盐酸多巴胺固体溶于1L的1.2M氢氧化钠水溶液中，在空气条件下(25℃，磁力搅拌24h)进行氧化反应来制备；
2)使用氢氧化钠溶液(1.2M)溶解葡聚糖，制备浓度为15％(w/v)的葡聚糖溶液；
3)取5mL步骤1中的盐酸多巴胺氢氧化钠水溶液和5mL步骤2中葡聚糖溶液混合在一起，室温搅拌均匀。
4)向步骤(3)中所制备的混合溶液中加入1.0mL交联剂EGDE，搅拌均匀后置于37℃恒温箱中，交联12h形成凝胶，记为D4。
实施例4：葡聚糖/聚多巴胺水凝胶支架的力学性能测试
其评估步骤如下：
设置对照组，使用氢氧化钠溶液(1.2M)溶解葡聚糖，制备浓度为15％(w/v)的葡聚糖溶液，取5mL葡聚糖溶液和5mL的1.2M的氢氧化钠溶液混合在一起，加入1.0mL交联剂EGDE，搅拌均匀后置于37℃恒温箱中，交联12h形成凝胶，记为D1。
通过使用流变仪对四组D1、D2、D3、D4不同多巴胺掺杂量的凝胶进行力学性能测试发现，随着聚多巴胺掺杂量的增加，复合凝胶的储能模量显著增加，由780Pa增加到1600Pa，其结果如图1所示。扫描电镜测试复合凝胶的表面形貌发现每种凝胶孔径均一，且孔径大小能够通过聚多巴胺掺杂量调节，孔径在5～30微米之间，如图2所示，随着聚多巴胺掺杂量的增加，孔径逐渐变小，对照组的凝胶孔隙最大，并且整体结构疏松，D2的孔径直径在15～25微米之间；D3的孔隙直径10-20微米之间；D4的孔隙直径在3-10微米之间，使其能够应用于不同细胞种类的支架，模拟不同细胞环境，具有更多样的应用。
实施例5：葡聚糖/聚多巴胺多孔水凝胶细胞支架与小鼠成纤维细胞共培养
其评估步骤如下：
设置对照组，使用氢氧化钠溶液(1.2M)溶解葡聚糖，制备浓度为15％(w/v)的葡聚糖溶液，取5mL葡聚糖溶液和5mL的1.2M的氢氧化钠溶液混合在一起，加入1.0mL交联剂EGDE，搅拌均匀后置于37℃恒温箱中，交联12h形成凝胶，记为D1。
取D1、D4的水凝胶，分成32份，每份干重约为5～7mg，放入孔板中75％酒精消毒12h，后用磷酸缓冲盐溶液(PBS)浸泡冲洗酒精1～2天，每天换液2～3次。
在共聚焦碟子中接种一定量的小鼠成纤维细胞(NIH3T3)，使用DMEM培养基，在5％CO2气体条件下37℃中孵育24h，之后加入D1、D4水凝胶样品，在成纤维细胞与水凝胶样品共培养1天、3天、5天后分别对共聚焦碟子中的成纤维细胞状态通过细胞活/死染色进行评价。实验中的细胞每两天换一次培养基。
图3结果显示，细胞伪足在纤维表明粘附延伸，细胞呈三角型、多角形以及梭型等多种形态。这意味着本发明者构筑的水凝胶多孔支架适合细胞粘附增殖和生长。相比于单纯的葡聚糖水凝胶(D1)，多巴胺掺杂的水凝胶(D4)在培养5天后细胞更加密集，说明多巴胺的掺杂可以有效促进细胞的粘附和增殖，该水凝胶支架为细胞的3D培养提供了一种新的可能。
实施例6：聚多巴胺掺杂的葡聚糖水凝胶支架用于伤口修复
其评估步骤如下：
设置对照组，使用氢氧化钠溶液(1.2M)溶解葡聚糖，制备浓度为15％(w/v)的葡聚糖溶液，取5mL葡聚糖溶液和5mL的1.2M的氢氧化钠溶液混合在一起，加入1.0mL交联剂EGDE，搅拌均匀后置于37℃恒温箱中，交联12h形成凝胶，记为D1。
将4月龄的SD大鼠随机分为3组，用戊巴比妥麻醉后，在鼠背部中间线皮肤处构建一个直径为8mm的圆形伤口。实验组在鼠背部中间贴上8mm大小的D4水凝胶薄片，空白对照组大鼠在伤口处贴上8mm大小的D1水凝胶薄片。术后观察时间点为0、7、14、21天，在对伤口愈合情况进行拍照记录后，采用戊巴比妥麻醉，对大鼠实施安乐死。
图4结果显示，相比于对照组(D1)，多巴胺掺杂的水凝胶组(D4)在培养21天后能有效地促进和加速大鼠的伤口愈合。本发明中所构筑的多巴胺掺水凝胶为临床医用伤口敷料提供了一种新的可能。且该水凝胶支架可以进一步的装载一些生长因子和抗菌药物等，使得这种材料具有修复皮肤伤口的潜在应用价值。
最后有必要在此说明的是：以上实施例只用于对本发明的技术方案作进一步详细地说明，不能理解为对本发明保护范围的限制，本领域的技术人员根据本发明的上述内容作出的一些非本质的改进和调整均属于本发明的保护范围。
Claims (7)
1.一种聚多巴胺掺杂葡聚糖水凝胶多孔支架的制备方法，其特征在于：将盐酸多巴胺固体溶于1.2M的氢氧化钠水溶液中，在常温条件下搅拌进行氧化反应制备含30mg/mL的盐酸多巴胺氢氧化钠水溶液，将葡聚糖溶于1.2M的氢氧化钠水溶液中，在常温条件下搅拌进行氧化反应制备含15％(w/v)葡聚糖溶液，将盐酸多巴胺氢氧化钠水溶液和葡聚糖溶液混合搅拌，加入与混合液体积比为1∶10的交联剂，所述盐酸多巴胺氢氧化钠水溶液和葡聚糖溶液体积比为1-5∶5。
2.一种聚多巴胺掺杂葡聚糖水凝胶多孔支架的制备方法，其特征在于：所述交联剂为乙二醇缩水甘油醚。
3.一种聚多巴胺掺杂葡聚糖水凝胶多孔支架，其特征在于，所述水凝胶多孔支架为根据权利要求1或2所述的一种聚多巴胺掺杂葡聚糖水凝胶多孔支架的制备方法制备的凝胶。
4.一种聚多巴胺掺杂葡聚糖水凝胶多孔支架的应用，其特征在于，根据权利要求3所述的一种聚多巴胺掺杂葡聚糖水凝胶多孔支架用于制备细胞支架的应用。
5.根据权利要求4所述的一种聚多巴胺掺杂葡聚糖水凝胶多孔支架的应用，其特征在于，所述细胞支架的孔隙为5～30μm。
6.一种聚多巴胺掺杂葡聚糖水凝胶多孔支架的应用，其特征在于，根据权利要求3所述的一种聚多巴胺掺杂葡聚糖水凝胶多孔支架用于制备皮肤修复凝胶的应用。
7.根据权利要求6所述一种聚多巴胺掺杂葡聚糖水凝胶多孔支架的应用，其特征在于，所述皮肤修复凝胶负载有细胞生长因子、抗菌剂、保湿剂、防腐剂、抗氧剂、乳化剂、增稠剂、甜味剂、胶浆剂、芳香剂以及矫味剂中的一种或多种。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010487754.XA CN111748120A (zh) | 2020-06-01 | 2020-06-01 | 一种聚多巴胺掺杂葡聚糖水凝胶多孔支架及制备方法及应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010487754.XA CN111748120A (zh) | 2020-06-01 | 2020-06-01 | 一种聚多巴胺掺杂葡聚糖水凝胶多孔支架及制备方法及应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111748120A true CN111748120A (zh) | 2020-10-09 |
Family
ID=72674189
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010487754.XA Pending CN111748120A (zh) | 2020-06-01 | 2020-06-01 | 一种聚多巴胺掺杂葡聚糖水凝胶多孔支架及制备方法及应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111748120A (zh) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113336967A (zh) * | 2021-05-13 | 2021-09-03 | 福州大学 | 一种pH响应的杀菌-自清洁水凝胶及其制备方法和应用 |
| CN113528424A (zh) * | 2021-08-25 | 2021-10-22 | 中国人民解放军陆军特色医学中心 | 一种光敏生物材料多孔支架及其应用 |
| CN113842494A (zh) * | 2021-09-10 | 2021-12-28 | 西北大学 | 一种促进组织再生的可注射止血晶胶及其制备方法和应用 |
| CN114230817A (zh) * | 2021-11-09 | 2022-03-25 | 东华大学 | 一种用于糖尿病足慢性伤口修复的水凝胶敷料及其制备方法 |
| CN115006585A (zh) * | 2022-06-01 | 2022-09-06 | 楼天女 | 一种止血性能好的伤口敷料及其制备方法 |
| CN115316405A (zh) * | 2022-09-14 | 2022-11-11 | 昆山博青生物科技有限公司 | 一种除味杀菌剂及其制备方法 |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100221303A1 (en) * | 2007-10-11 | 2010-09-02 | Universite Paris 7 - Denis Diderot | Method for Preparing Porous Scaffold for Tissue Engineering, Cell Culture and Cell Delivery |
| CN110237782A (zh) * | 2019-06-26 | 2019-09-17 | 中南民族大学 | 一种高强度抗氧化壳聚糖/聚多巴胺复合水凝胶的制备方法 |
| CN111068627A (zh) * | 2019-11-29 | 2020-04-28 | 温州医科大学 | 一种聚多巴胺/普鲁兰复合水凝胶以及制备方法和应用 |
-
2020
- 2020-06-01 CN CN202010487754.XA patent/CN111748120A/zh active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100221303A1 (en) * | 2007-10-11 | 2010-09-02 | Universite Paris 7 - Denis Diderot | Method for Preparing Porous Scaffold for Tissue Engineering, Cell Culture and Cell Delivery |
| CN110237782A (zh) * | 2019-06-26 | 2019-09-17 | 中南民族大学 | 一种高强度抗氧化壳聚糖/聚多巴胺复合水凝胶的制备方法 |
| CN111068627A (zh) * | 2019-11-29 | 2020-04-28 | 温州医科大学 | 一种聚多巴胺/普鲁兰复合水凝胶以及制备方法和应用 |
Non-Patent Citations (1)
| Title |
|---|
| 姜锡瑞等: "《生物发酵产业技术》", 31 May 2016, 北京：中国轻工业出版社 * |
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113336967A (zh) * | 2021-05-13 | 2021-09-03 | 福州大学 | 一种pH响应的杀菌-自清洁水凝胶及其制备方法和应用 |
| CN113336967B (zh) * | 2021-05-13 | 2023-03-24 | 福州大学 | 一种pH响应的杀菌-自清洁水凝胶及其制备方法和应用 |
| CN113528424A (zh) * | 2021-08-25 | 2021-10-22 | 中国人民解放军陆军特色医学中心 | 一种光敏生物材料多孔支架及其应用 |
| CN113842494A (zh) * | 2021-09-10 | 2021-12-28 | 西北大学 | 一种促进组织再生的可注射止血晶胶及其制备方法和应用 |
| CN114230817A (zh) * | 2021-11-09 | 2022-03-25 | 东华大学 | 一种用于糖尿病足慢性伤口修复的水凝胶敷料及其制备方法 |
| CN115006585A (zh) * | 2022-06-01 | 2022-09-06 | 楼天女 | 一种止血性能好的伤口敷料及其制备方法 |
| CN115316405A (zh) * | 2022-09-14 | 2022-11-11 | 昆山博青生物科技有限公司 | 一种除味杀菌剂及其制备方法 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111748120A (zh) | 一种聚多巴胺掺杂葡聚糖水凝胶多孔支架及制备方法及应用 | |
| Mi et al. | In vitro evaluation of a chitosan membrane cross-linked with genipin | |
| US11511018B2 (en) | Conductive biomimetic skin scaffold material with self-repairing function and a method of preparing the same | |
| CN104761737B (zh) | 一种静电纺丝法制备胶原蛋白/氧化石墨烯纳米纤维复合膜的方法 | |
| CN111840631A (zh) | 可注射型抗菌止血水凝胶粘合剂及其制备方法和应用 | |
| Ul-Islam et al. | Ex situ synthesis and characterization of high strength multipurpose bacterial cellulose-aloe vera hydrogels | |
| US20220333080A1 (en) | Crosslinked Hydrogel for Muscle Stem Cell Culture and Preparation Method and Use Thereof | |
| CN110698866A (zh) | 一种超声介导丝素蛋白复合胶原水凝胶及其制备方法 | |
| WO2023143540A1 (zh) | 一种基于单向纳米孔脱水的功能性高分子膜/水凝胶膜、制备方法及装置 | |
| Hashemi et al. | Polyurethane/chitosan/hyaluronic acid scaffolds: providing an optimum environment for fibroblast growth | |
| CN105176095B (zh) | 一种胶原基有机硅橡胶多孔复合膜及其制备方法 | |
| Andleeb et al. | Hydrogel patch with pretreated stem cells accelerates wound closure in diabetic rats | |
| CN110452413B (zh) | 一种胶原蛋白交联剂组合物及其应用 | |
| CN109943974B (zh) | 基于聚羟基脂肪酸酯/明胶电纺纳米纤维的神经导管材料的制备方法 | |
| CN115501381B (zh) | 替代可吸收缝合线的脂肪组织粘合剂、制备方法及其应用 | |
| Zhang et al. | A bacteriostatic and hemostatic medical dressing based on PEG modified keratin/carboxymethyl chitosan | |
| CN113354803B (zh) | 一种含醛基侧基的聚碳酸酯/聚乙二醇嵌段共聚物、可注射自修复水凝胶敷料及制备方法 | |
| Kang et al. | Fabrication and characterization of an electro‐compacted collagen/elastin/hyaluronic acid sheet as a potential skin scaffold | |
| Liu et al. | Reinforcement of a porous collagen scaffold with surface-activated PLA fibers | |
| CN115322451A (zh) | 一种纳米羟基磷灰石复合水凝胶及其制备方法和应用 | |
| CN115487358A (zh) | 一种用于软骨组织修复的凝胶复合支架及制备方法 | |
| CN113018517A (zh) | 一种3d打印皮肤支架及其制备方法和应用 | |
| Denost et al. | Colorectal wall regeneration resulting from the association of chitosan hydrogel and stromal vascular fraction from adipose tissue | |
| He et al. | Evaluation of biological properties of collagen/hyaluronic acid composite scaffolds | |
| CN117964918A (zh) | 一种可注射酶交联的酪胺修饰的蛋清蛋白水凝胶的制备方法 |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201009 |
|
| RJ01 | Rejection of invention patent application after publication |