CN112980003A - 一种基于天然多糖型抗菌水凝胶、制备方法及用途 - Google Patents
一种基于天然多糖型抗菌水凝胶、制备方法及用途 Download PDFInfo
- Publication number
- CN112980003A CN112980003A CN202110416389.8A CN202110416389A CN112980003A CN 112980003 A CN112980003 A CN 112980003A CN 202110416389 A CN202110416389 A CN 202110416389A CN 112980003 A CN112980003 A CN 112980003A
- Authority
- CN
- China
- Prior art keywords
- gox
- chitosan
- hyaluronic acid
- preparation
- natural polysaccharide
- 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 54
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 23
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- 150000004676 glycans Chemical class 0.000 title claims abstract description 18
- 229920001282 polysaccharide Polymers 0.000 title claims abstract description 18
- 239000005017 polysaccharide Substances 0.000 title claims abstract description 18
- 230000002757 inflammatory effect Effects 0.000 claims abstract description 7
- 230000035876 healing Effects 0.000 claims abstract description 6
- 239000000463 material Substances 0.000 claims abstract description 3
- 229920001661 Chitosan Polymers 0.000 claims description 29
- 239000004366 Glucose oxidase Substances 0.000 claims description 25
- 229940116332 glucose oxidase Drugs 0.000 claims description 25
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 17
- 229920002674 hyaluronan Polymers 0.000 claims description 17
- 229960003160 hyaluronic acid Drugs 0.000 claims description 17
- 238000006243 chemical reaction Methods 0.000 claims description 15
- 239000000243 solution Substances 0.000 claims description 13
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 13
- 239000008367 deionised water Substances 0.000 claims description 10
- 229910021641 deionized water Inorganic materials 0.000 claims description 10
- 108010015776 Glucose oxidase Proteins 0.000 claims description 9
- 150000001875 compounds Chemical class 0.000 claims description 9
- 235000019420 glucose oxidase Nutrition 0.000 claims description 9
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 7
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 6
- 239000002840 nitric oxide donor Substances 0.000 claims description 6
- JQWHASGSAFIOCM-UHFFFAOYSA-M sodium periodate Chemical compound [Na+].[O-]I(=O)(=O)=O JQWHASGSAFIOCM-UHFFFAOYSA-M 0.000 claims description 6
- 239000004475 Arginine Substances 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical compound NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 claims description 4
- 239000002262 Schiff base Substances 0.000 claims description 4
- 150000004753 Schiff bases Chemical class 0.000 claims description 4
- 238000007112 amidation reaction Methods 0.000 claims description 4
- 125000003277 amino group Chemical group 0.000 claims description 4
- 239000007864 aqueous solution Substances 0.000 claims description 4
- BPMFZUMJYQTVII-UHFFFAOYSA-N guanidinoacetic acid Chemical compound NC(=N)NCC(O)=O BPMFZUMJYQTVII-UHFFFAOYSA-N 0.000 claims description 4
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 229960000583 acetic acid Drugs 0.000 claims description 3
- 230000002378 acidificating effect Effects 0.000 claims description 3
- 125000003172 aldehyde group Chemical group 0.000 claims description 3
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 3
- 239000003814 drug Substances 0.000 claims description 3
- 229940088598 enzyme Drugs 0.000 claims description 3
- 239000002243 precursor Substances 0.000 claims description 3
- MEJYXFHCRXAUIL-UHFFFAOYSA-N 2-[carbamimidoyl(methyl)amino]acetic acid;hydrate Chemical compound O.NC(=N)N(C)CC(O)=O MEJYXFHCRXAUIL-UHFFFAOYSA-N 0.000 claims description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 2
- CHJJGSNFBQVOTG-UHFFFAOYSA-N N-methyl-guanidine Natural products CNC(N)=N CHJJGSNFBQVOTG-UHFFFAOYSA-N 0.000 claims description 2
- 150000001413 amino acids Chemical class 0.000 claims description 2
- 238000010523 cascade reaction Methods 0.000 claims description 2
- 229960004826 creatine monohydrate Drugs 0.000 claims description 2
- 238000004132 cross linking Methods 0.000 claims description 2
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 claims description 2
- 239000012362 glacial acetic acid Substances 0.000 claims description 2
- 230000001737 promoting effect Effects 0.000 claims description 2
- 230000008439 repair process Effects 0.000 claims description 2
- ZRALSGWEFCBTJO-UHFFFAOYSA-O guanidinium Chemical compound NC(N)=[NH2+] ZRALSGWEFCBTJO-UHFFFAOYSA-O 0.000 claims 3
- 238000000502 dialysis Methods 0.000 claims 1
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 abstract description 48
- 210000004204 blood vessel Anatomy 0.000 abstract description 4
- 238000010606 normalization Methods 0.000 abstract description 4
- 238000011160 research Methods 0.000 abstract description 3
- 241000894006 Bacteria Species 0.000 abstract description 2
- 206010052428 Wound Diseases 0.000 description 15
- 208000027418 Wounds and injury Diseases 0.000 description 15
- 230000029663 wound healing Effects 0.000 description 10
- 230000001954 sterilising effect Effects 0.000 description 6
- 238000004659 sterilization and disinfection Methods 0.000 description 6
- 238000005303 weighing Methods 0.000 description 6
- 238000004108 freeze drying Methods 0.000 description 5
- 206010072170 Skin wound Diseases 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 description 3
- 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 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 229910052739 hydrogen Inorganic materials 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 2
- 206010063560 Excessive granulation tissue Diseases 0.000 description 2
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 241000191967 Staphylococcus aureus Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 210000001126 granulation tissue Anatomy 0.000 description 2
- 125000002795 guanidino group Chemical group C(N)(=N)N* 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 238000003756 stirring Methods 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 description 1
- 102000008186 Collagen Human genes 0.000 description 1
- 108010035532 Collagen Proteins 0.000 description 1
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 description 1
- 108010001742 S-Nitrosoglutathione Proteins 0.000 description 1
- HYHSBSXUHZOYLX-WDSKDSINSA-N S-nitrosoglutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CSN=O)C(=O)NCC(O)=O HYHSBSXUHZOYLX-WDSKDSINSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006502 antiplatelets effects Effects 0.000 description 1
- 230000003305 autocrine Effects 0.000 description 1
- 125000005615 azonium group Chemical group 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 229920001436 collagen Polymers 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 210000002950 fibroblast Anatomy 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000012737 fresh medium Substances 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001879 gelation Methods 0.000 description 1
- 239000003102 growth factor Substances 0.000 description 1
- 239000000416 hydrocolloid Substances 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000005012 migration Effects 0.000 description 1
- 238000013508 migration Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 230000037311 normal skin Effects 0.000 description 1
- 238000000655 nuclear magnetic resonance spectrum Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 229940082615 organic nitrates used in cardiac disease Drugs 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000003076 paracrine Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 210000000578 peripheral nerve Anatomy 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000004381 surface treatment Methods 0.000 description 1
- 230000030968 tissue homeostasis Effects 0.000 description 1
- 238000011282 treatment Methods 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 230000024883 vasodilation Effects 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
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
- A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/42—Proteins; Polypeptides; Degradation products thereof; Derivatives thereof, e.g. albumin, gelatin or zein
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/56—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule
- A61K47/61—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic macromolecular compound, e.g. an oligomeric, polymeric or dendrimeric molecule the organic macromolecular compound being a polysaccharide or a derivative thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/02—Local antiseptics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial 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
- C08J3/00—Processes of treating or compounding macromolecular substances
- C08J3/24—Crosslinking, e.g. vulcanising, of macromolecules
-
- 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
- C08J2305/08—Chitin; Chondroitin sulfate; Hyaluronic acid; 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
- C08J2405/00—Characterised by the use of polysaccharides or of their derivatives not provided for in groups C08J2401/00 or C08J2403/00
- C08J2405/08—Chitin; Chondroitin sulfate; Hyaluronic acid; Derivatives thereof
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Epidemiology (AREA)
- General Chemical & Material Sciences (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Dispersion Chemistry (AREA)
- Polymers & Plastics (AREA)
- Inorganic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Dermatology (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明公开了一种基于天然多糖型抗菌水凝胶、制备方法及其应用，该水凝胶是由胍基化壳聚糖和葡萄糖氧化酶接枝的透明质酸为主要材料，配制成不同比例的混合溶液，通过席夫碱交联反应制备而成。本发明的抗菌水凝胶材料在炎症伤口处程序性释放一氧化氮，抑制细菌生长，同时促进血管正常化，加速伤口愈合。抗菌水凝胶制备方法简单，使用方便，具有良好的研究和开发利用前景。
Description
技术领域
本发明涉及化工医药，特别涉及一种基于天然多糖型抗菌水凝胶、制备方法及其应用。
背景技术
皮肤覆盖全身表面，不仅是人体最大的器官之一，而且还是维持体内环境稳定和抵御外界环境感染的重要屏障。由于皮肤是直接与外界环境相接触的组织，大面积暴露于外界，使得皮肤很容易遭受外力作用造成皮肤组织出现离断或缺损，导致皮肤创面的形成。而在皮肤创面愈合过程中，伤口极易受到大肠杆菌和金黄色葡萄球菌等病原体的感染，引起感染性炎症，导致伤口难以愈合。因此，探究皮肤炎性伤口愈合的机制，为皮肤创伤疾病提供新的治疗策略，具有重大意义。
传统的创伤治疗方法是将伤口经过杀菌清洗，用纱布包裹，但是纱布易于皮肤伤口组织粘连，换药时常常破坏新生的上皮和肉芽组织，引起出血，造成病人二次疼痛。Winter课题组在1962年提出“湿性愈合”的理论，Hinman和Maibach也论证了湿性环境相比干燥环境，可以显著提高伤口愈合的速度。从而奠定了采用新型敷料处理创面的理论基础。基于这一思想的各种新式敷料得到广泛的发展，如水状胶体敷料、水凝胶敷料等。其中水凝胶敷料可以保持伤口局部湿润，有助于血管的正常化；同时可以保护新生肉芽组织不受损伤；保护末梢神经，减少疼痛；可以促进生长因子释放，促进创面的愈合。因此得到各国的普遍重视。
近年来将一氧化氮(NO)应用于创伤愈合这一理念受到了极大的关注与重视，NO与NO供体如有机硝酸酯类、偶氮鎓二醇盐、S-亚硝基谷胱甘肽及载有NO的纳米粒子均被证实可以在不同程度上促进创伤愈合。越来越多的研究表明NO作为一种关键的自分泌与旁分泌因子在维持正常皮肤组织内环境稳态中发挥着重要作用。在创伤愈合的三个阶段中，NO分别扮演着不同的作用：首先，在炎症阶段，NO介导血管舒张和抗血小板作用；其次在增殖期，NO促进新生血管正常化，成纤维细胞、上皮细胞和内皮细胞的迁移、增殖；最后在重塑步骤里，NO能够加速创面胶原形成及增强创面组织机械强度。因此，NO应用于伤口愈合方面具有巨大的研究价值。
发明内容
发明目的：为了解决皮肤创面受细菌感染，引发炎症导致伤口难以愈合的问题，提供一种具有较强的杀菌抑菌效果的基于天然多糖型抗菌水凝胶、
本发明另一目的是提供所述抗菌水凝胶的制备方法及其用途。
技术方案：本发明提供一种基于天然多糖型抗菌水凝胶，由胍基化壳聚糖(G-CS)和透明质酸-葡萄糖氧化酶(OHA-GOx)通过席夫碱交联反应制备而成。
进一步地，所述胍基化壳聚糖(G-CS)是由壳聚糖(CS)上氨基通过酰胺化反应修饰含有羧基的胍基型NO供体化合物获得。
进一步地，所述含有羧基的胍基型NO供体化合物为精氨酸(Arg)、胍基乙酸或一水肌酸等氨基酸及其衍生物。
进一步地，所述透明质酸-葡萄糖氧化酶(OHA-GOx)是由氧化后带有醛基结构的透明质酸(HA)的部分醛基通过席夫碱反应与葡萄糖氧化酶(GOx)上的氨基连接得到OHA-GOx。
进一步地，所述胍基化壳聚糖(G-CS)和透明质酸-葡萄糖氧化酶(OHA-GOx)的质量比为：10∶1-1∶1，优选为7∶1-1∶1。
所述基于天然多糖型抗菌水凝胶制备方法包括以下步骤：
(1)胍基化壳聚糖(G-CS)的制备
称取壳聚糖(CS)完全溶解于微酸水溶液中，再称取含有羧基的胍基型NO供体化合物，通过酰胺化反应过夜，经透析冻干得到胍基化壳聚糖(G-CS)。
(2)透明质酸-葡萄糖氧化酶(OHA-GOx)的制备
称取透明质酸(HA)和高碘酸钠溶解于水中，室温下反应48h，加入终止剂终止反应，得到的反应液透析冻干，然后称取冻干产物与GOx，室温下在去离子水中避光反应，透析冻干得到OHA-GOx。
(3)水凝胶的制备
将步骤(1)和(2)得到的胍基化壳聚糖(G-CS)和透明质酸-葡萄糖氧化酶(OHA-GOx)溶于PBS，通过一定质量比混合均匀，静置得到目标产物天然多糖型抗菌水凝胶。
(3)将两种CAHG水凝胶前体溶液，分装于喷瓶中，在皮肤患者创面处喷射原位成胶，利用炎症伤口处含糖环境来进行程序性的NO的释放，进行杀菌抗菌，促进伤口愈合。
进一步地，步骤(1)中微酸水溶液可选择冰乙酸或稀盐酸。
进一步地，步骤(1)壳聚糖(CS)分子量为30-1000kDa，步骤(2)透明质酸(HA)分子量为50-1200kDa。
进一步地，步骤(3)中水凝胶前体材料溶解采用去离子水、PB或PBS。
所述抗菌水凝胶在制备炎症伤口处利用酶级联反应促进修复愈合药物中的用途。
有益效果：本发明通过程序性的释放NO和H2O2协同杀菌促进伤口愈合，采用的喷射型原位成胶给药方式，便于创面治疗；本发明制备的水凝胶具有较强的杀菌抑菌效果，利用炎症伤口处含糖环境程序性的释放NO和H2O2，来达到协同杀菌抑菌，同时NO能促进血管正常化，加速伤口愈合；另一方面，该过程主要原材料为天然多糖，来源广泛、储量丰富，且制备方法操作简单、条件温和，适合工业化生产，具有广阔的应用前景。
附图说明
图1实施例1中CS-Arg的氢核磁图谱；
图2实施例2中OHA的氢核磁图谱；
图3实施例3中CAHG水凝胶不同比例的成胶图谱；
图4实施例4中CAHG水凝胶的H2O2的释放图谱；
图5实施例4中CAHG水凝胶的NO的释放图谱；
图6实施例5中CAHG水凝胶的体外抑菌图谱。
具体实施方式
实施例1：壳聚糖-精氨酸(CS-Arg)的制备
CS-Arg的合成，合成路线及过程如下：
称取0.5g壳聚糖(CS)于反应瓶，加入50mL的醋酸溶液；另取1.0g精氨酸以及1.3gN-羟基琥珀酰亚胺(NHS)和3.3g(1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐)(EDC)溶于10mL去离子水中，加入完全溶解后的壳聚糖溶液中，调节pH为4-6，室温下磁力搅拌反应过夜。反应结束后，将反应液用去离子水透析48h，冻干得到化合物壳聚糖-精氨酸(CS-Arg)。
实施例2：透明质酸-葡萄糖氧化酶(OHA-GOx)的制备
OHA-GOx的合成，合成路线及过程如下：
称取1.0g透明质酸(HA)和1.4g高碘酸钠溶解在250mL去离子水中，室温避光下磁力搅拌48h，反应结束加入1mL的乙二醇终止反应，得到的反应液再经过透析24h，冻干得到化合物氧化的透明质酸(OHA)。
称取100mg冻干好的OHA溶解于50mL去离子水中，再称取10mg葡萄糖氧化酶(GOx)，溶解在10mL去离子水中，然后将其缓慢滴加到OHA溶液里。室温下避光磁力搅拌24h，得到的反应液再透析24h，冻干得到化合物氧化的透明质酸-葡萄糖氧化酶(OHA-GOx)。
实施例3：水凝胶的制备
称取60mg CS-Arg和100mg OHA-GOx，分别溶于1mL PBS里，得到60mg/mL的CS-Arg溶液以及100mg/mL的OHA-GOx溶液，并以质量比3∶1混合，即得到CAHG水凝胶。如图3，CS-Arg和OHA-GOx在1∶1、2∶1和3∶1质量比下的倒置成胶情况，在1∶1时，混合溶液还是流动态，未完全成胶，当CS-Arg的比例提高为3倍时，倒置摆放30秒不流动，为凝胶态。
实施例4：CAHG水凝胶的H2O2和NO体外释放。
NO和H2O2的体外释放实验在37℃下，两种不同介质：(i)PBS缓冲液；(ii)PBS缓冲液(含1mg/mL葡萄糖)。制备好的CAHG水凝胶样品分成两份，将其置于20mL的相应缓冲液中，然后放入37℃恒温摇床。在指定的时间点，从释放体系取出5mL的释放介质，然后补充相同体积的新鲜介质。释放的H2O2和NO用对应的检测试剂盒染色后用酶标仪测定。该释放实验重复三次。如图4、5所示，由CAHG水凝胶的H2O2和NO释放情况可知，在含葡萄糖条件下，CAHG水凝胶能利用酶级联反应程序性的释放H2O2和NO，而CAHG水凝胶在PBS缓冲液里时，H2O2基本不释放，NO释放量大大减少。
实施例5：CAHG水凝胶的体外抑菌圈实验
用灭菌水溶解制备好相同比例浓度的壳聚糖-氧化透明质酸水凝胶(CH水凝胶)、壳聚糖-精氨酸-氧化透明质酸水凝胶(CAH水凝胶)、壳聚糖-氧化透明质酸-葡萄糖氧化酶水凝胶(CHG水凝胶)和壳聚糖-精氨酸-氧化透明质酸-葡萄糖氧化酶水凝胶(CAHG水凝胶)等四组水凝胶待用；大肠杆菌和金黄色葡萄球菌经LB液体培养基培养至对数期并测量OD600≈0.2，细菌浓度为3×108CFU/mL，取20uL菌液用涂布棒均匀涂布在LB固体培养基表面，再用挖孔器在固体培养基上挖出一个直径5mm的孔，将制备好的四组水凝胶置于孔中，最终将培养基置于37℃恒温箱里培养12h后观察抑菌圈直径大小。如图5，四种水凝胶的抑菌实验表明，CAHG水凝胶相对于其他四组表现出了显著的抗菌效果，说明H2O2和NO在含有葡萄糖的环境下能释放出来达到杀菌抑菌的作用。
Claims (10)
1.一种基于天然多糖型抗菌水凝胶，其特征在于，由胍基化壳聚糖(G-CS)和透明质酸-葡萄糖氧化酶(OHA-GOx)通过席夫碱交联反应制备而成。
2.根据权利要求1所述的基于天然多糖型抗菌水凝胶，其特征在于，所述胍基化壳聚糖(G-CS)是由壳聚糖(CS)上氨基通过酰胺化反应修饰含有羧基的胍基型NO供体化合物获得。
3.根据权利要求1所述的基于天然多糖型抗菌水凝胶，其特征在于，所述含有羧基的胍基型NO供体化合物为精氨酸(Arg)、胍基乙酸或一水肌酸等氨基酸及其衍生物。
4.根据权利要求1所述的基于天然多糖型抗菌水凝胶，其特征在于，所述透明质酸-葡萄糖氧化酶(OHA-GOx)是由氧化后带有醛基结构的透明质酸(HA)的部分醛基通过席夫碱反应与葡萄糖氧化酶(GOx)上的氨基连接得到OHA-GOx。
5.根据权利要求1所述的基于天然多糖型抗菌水凝胶，其特征在于，所述胍基化壳聚糖(G-CS)和透明质酸-葡萄糖氧化酶(OHA-GOx)的质量比为：10∶1-1∶1。
6.权利要求1-5任一项所述基于天然多糖型抗菌水凝胶的制备方法，其特征在于，包括以下步骤：
(1)胍基化壳聚糖(G-CS)的制备：
称取壳聚糖(CS)完全溶解于微酸水溶液中，再称取含有羧基的胍基型NO供体化合物，通过酰胺化反应过夜，经透析冻干得到胍基化壳聚糖(G-CS)；
(2)透明质酸-葡萄糖氧化酶(OHA-GOx)的制备：
称取透明质酸(HA)和高碘酸钠溶解于去离子水里，室温下反应，加入终止剂终止反应，得到的反应液透析冻干，然后称取冻干产物与葡萄糖氧化酶(GOx)，室温下在去离子水中避光反应，透析冻干得到透明质酸-葡萄糖氧化酶(OHA-GOx)；
(3)水凝胶的制备：
将步骤(1)和(2)得到的胍基化壳聚糖(G-CS)和透明质酸-葡萄糖氧化酶(OHA-GOx)溶解，通过一定质量比混合均匀，静置，即得。
7.根据权利要求6所述基于天然多糖型抗菌水凝胶的制备方法，其特征在于，步骤(1)中微酸水溶液可选择冰乙酸或稀盐酸。
8.根据权利要求6所述基于天然多糖型抗菌水凝胶的制备方法，其特征在于，步骤(1)壳聚糖(CS)分子量为30-1000kDa，步骤(2)透明质酸(HA)分子量为50-1200kDa。
9.根据权利要求6所述基于天然多糖型抗菌水凝胶的制备方法，其特征在于，步骤(3)中水凝胶前体材料溶解采用去离子水、PB或PBS。
10.权利要求1所述抗菌水凝胶在制备炎症伤口处利用酶级联反应促进修复愈合药物中的用途。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110416389.8A CN112980003B (zh) | 2021-04-16 | 2021-04-16 | 一种基于天然多糖型抗菌水凝胶、制备方法及用途 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110416389.8A CN112980003B (zh) | 2021-04-16 | 2021-04-16 | 一种基于天然多糖型抗菌水凝胶、制备方法及用途 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112980003A true CN112980003A (zh) | 2021-06-18 |
| CN112980003B CN112980003B (zh) | 2022-06-28 |
Family
ID=76341010
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110416389.8A Active CN112980003B (zh) | 2021-04-16 | 2021-04-16 | 一种基于天然多糖型抗菌水凝胶、制备方法及用途 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112980003B (zh) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114010584A (zh) * | 2021-11-09 | 2022-02-08 | 华熙生物科技股份有限公司 | 一种抑菌创面修复凝胶及其制备方法 |
| CN114716694A (zh) * | 2022-05-06 | 2022-07-08 | 南京工业大学 | 一种促血管化和抗钙化且可3d打印心脏瓣膜的水凝胶 |
| CN115725091A (zh) * | 2022-11-28 | 2023-03-03 | 中国药科大学 | 响应性控释一氧化氮光热水凝胶及其制备方法和应用 |
| CN116082717A (zh) * | 2023-02-02 | 2023-05-09 | 领博生物科技(杭州)有限公司 | 水凝胶、血管纤维骨架及其制备方法和应用 |
| CN118812877A (zh) * | 2024-06-17 | 2024-10-22 | 弘知生物科技(浙江)有限公司 | 一种含聚胍类高分子聚合物的长效抑菌水凝胶及制备方法 |
Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004112851A1 (en) * | 2003-06-23 | 2004-12-29 | Johnson & Johnson Medical Limited | Hydrogel compositions comprising enzymes |
| WO2010037395A2 (en) * | 2008-10-01 | 2010-04-08 | Dako Denmark A/S | Mhc multimers in cancer vaccines and immune monitoring |
| WO2010037397A1 (en) * | 2008-10-01 | 2010-04-08 | Dako Denmark A/S | Mhc multimers in cmv immune monitoring |
| US20100168390A1 (en) * | 2007-07-03 | 2010-07-01 | Dako Denmark A/S | Mhc multimers, methods for their generation, labeling and use |
| CN105726463A (zh) * | 2015-12-09 | 2016-07-06 | 宁波大学 | 一种用于胰岛素精控释放的双响应双交联可注射水凝胶的制备方法及应用 |
| US20170049111A1 (en) * | 2014-04-30 | 2017-02-23 | Matoke Holdings Limited | Antimicrobial compositions |
| US20190177683A1 (en) * | 2017-12-07 | 2019-06-13 | Indiana University Research And Technology Corporation | Stabilization of enzyme-immobilized hydrogels for extended hypoxic cell culture |
| US20190307904A1 (en) * | 2016-03-30 | 2019-10-10 | Convatec Technologies Inc. | Modified wound dressings |
| CN111154149A (zh) * | 2019-12-31 | 2020-05-15 | 广州医科大学附属第一医院（广州呼吸中心） | 一种水凝胶及其制备方法与敷料 |
| CN111410754A (zh) * | 2020-03-30 | 2020-07-14 | 暨南大学 | 一种s-亚硝基硫醇/多糖基原位成型水凝胶及其制备方法与应用 |
| CN111870693A (zh) * | 2020-08-12 | 2020-11-03 | 中国药科大学 | 一种黑磷促进精氨酸快速释放no的方法 |
-
2021
- 2021-04-16 CN CN202110416389.8A patent/CN112980003B/zh active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2004112851A1 (en) * | 2003-06-23 | 2004-12-29 | Johnson & Johnson Medical Limited | Hydrogel compositions comprising enzymes |
| US20100168390A1 (en) * | 2007-07-03 | 2010-07-01 | Dako Denmark A/S | Mhc multimers, methods for their generation, labeling and use |
| WO2010037395A2 (en) * | 2008-10-01 | 2010-04-08 | Dako Denmark A/S | Mhc multimers in cancer vaccines and immune monitoring |
| WO2010037397A1 (en) * | 2008-10-01 | 2010-04-08 | Dako Denmark A/S | Mhc multimers in cmv immune monitoring |
| US20170049111A1 (en) * | 2014-04-30 | 2017-02-23 | Matoke Holdings Limited | Antimicrobial compositions |
| CN105726463A (zh) * | 2015-12-09 | 2016-07-06 | 宁波大学 | 一种用于胰岛素精控释放的双响应双交联可注射水凝胶的制备方法及应用 |
| US20190307904A1 (en) * | 2016-03-30 | 2019-10-10 | Convatec Technologies Inc. | Modified wound dressings |
| US20190177683A1 (en) * | 2017-12-07 | 2019-06-13 | Indiana University Research And Technology Corporation | Stabilization of enzyme-immobilized hydrogels for extended hypoxic cell culture |
| CN111154149A (zh) * | 2019-12-31 | 2020-05-15 | 广州医科大学附属第一医院（广州呼吸中心） | 一种水凝胶及其制备方法与敷料 |
| CN111410754A (zh) * | 2020-03-30 | 2020-07-14 | 暨南大学 | 一种s-亚硝基硫醇/多糖基原位成型水凝胶及其制备方法与应用 |
| CN111870693A (zh) * | 2020-08-12 | 2020-11-03 | 中国药科大学 | 一种黑磷促进精氨酸快速释放no的方法 |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114010584A (zh) * | 2021-11-09 | 2022-02-08 | 华熙生物科技股份有限公司 | 一种抑菌创面修复凝胶及其制备方法 |
| CN114010584B (zh) * | 2021-11-09 | 2023-10-20 | 华熙生物科技股份有限公司 | 一种抑菌创面修复凝胶及其制备方法 |
| CN114716694A (zh) * | 2022-05-06 | 2022-07-08 | 南京工业大学 | 一种促血管化和抗钙化且可3d打印心脏瓣膜的水凝胶 |
| CN115725091A (zh) * | 2022-11-28 | 2023-03-03 | 中国药科大学 | 响应性控释一氧化氮光热水凝胶及其制备方法和应用 |
| CN116082717A (zh) * | 2023-02-02 | 2023-05-09 | 领博生物科技(杭州)有限公司 | 水凝胶、血管纤维骨架及其制备方法和应用 |
| CN118812877A (zh) * | 2024-06-17 | 2024-10-22 | 弘知生物科技(浙江)有限公司 | 一种含聚胍类高分子聚合物的长效抑菌水凝胶及制备方法 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN112980003B (zh) | 2022-06-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112980003B (zh) | 一种基于天然多糖型抗菌水凝胶、制备方法及用途 | |
| CN110314242B (zh) | 一种可控释放的抗生素复合水凝胶的制备方法及其用途 | |
| CN105597146B (zh) | 伤口愈合用多功能温敏凝胶复合物敷料及其制备与使用方法 | |
| CN101654529B (zh) | 壳聚糖和/或其衍生物金属复合物的制备方法及应用 | |
| CN112480434B (zh) | 一种铜离子抗菌水凝胶及制备方法和应用 | |
| CN112851951B (zh) | 一种接枝ε-聚赖氨酸的双醛基壳聚糖及其制备方法和应用 | |
| CN103159967B (zh) | 一种具有自抗炎功能的胶原基海绵伤口敷料的制备方法 | |
| WO2023231049A1 (zh) | 一种促进创面无瘢痕愈合的抗菌敷料及其制备方法 | |
| CN111494709A (zh) | 兼具抗肿瘤和抑菌功能的促组织修复水凝胶的制备及应用 | |
| CN105837837B (zh) | 一种含有纳米银的医用壳聚糖温敏水凝胶的制备方法 | |
| TWI478943B (zh) | 高分子組成物用於製備具有抑制基質金屬蛋白酶之活性的功效的醫療器材或抑制劑的用途 | |
| CN109758604B (zh) | 一种皮肤修复制剂及其制备方法和应用 | |
| CN102648987B (zh) | 一种不对称双交联复合材料及其制备方法和应用 | |
| CN104740141B (zh) | 一种抗菌喷剂及其制备方法 | |
| CN114456239A (zh) | 生泰素以及由其制备的外用抗菌肽凝胶制剂与应用 | |
| CN115869459A (zh) | 促伤口愈合的多肽水凝胶及其制备方法与应用 | |
| CN113509591A (zh) | 一种抗菌阳离子可注射水凝胶敷料及其制备方法 | |
| CN111410754A (zh) | 一种s-亚硝基硫醇/多糖基原位成型水凝胶及其制备方法与应用 | |
| CN107049930A (zh) | 一种促愈合伤口凝胶及其制备方法 | |
| CN114479124B (zh) | 一种可自愈水凝胶、其制备方法及应用 | |
| CN116920165A (zh) | 一种原位产氧的水凝胶及其制备方法和应用 | |
| CN116376123A (zh) | 可注射葡聚糖基水凝胶、负载重楼皂苷i的葡聚糖基水凝胶及制备方法与应用 | |
| CN115850731A (zh) | 一种可注射多功能糖肽水凝胶、制备方法及其应用 | |
| CN112618785B (zh) | 一种多孔抗菌水凝胶敷料及其制备方法 | |
| CN115558128A (zh) | 一种具有过氧化物酶活性水凝胶敷料及其制备方法和应用 |
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 |