CN103272268B - 一种抗菌角膜修复材料及其制备方法 - Google Patents
一种抗菌角膜修复材料及其制备方法 Download PDFInfo
- Publication number
- CN103272268B CN103272268B CN201310181772.5A CN201310181772A CN103272268B CN 103272268 B CN103272268 B CN 103272268B CN 201310181772 A CN201310181772 A CN 201310181772A CN 103272268 B CN103272268 B CN 103272268B
- Authority
- CN
- China
- Prior art keywords
- cornea
- solution
- cross
- collagen
- antibacterial
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 239000000463 material Substances 0.000 title claims abstract description 58
- 210000004087 cornea Anatomy 0.000 title claims abstract description 42
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 19
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 229920001436 collagen Polymers 0.000 claims abstract description 35
- 239000000243 solution Substances 0.000 claims abstract description 35
- 102000008186 Collagen Human genes 0.000 claims abstract description 31
- 108010035532 Collagen Proteins 0.000 claims abstract description 31
- 239000012528 membrane Substances 0.000 claims abstract description 25
- 238000004132 cross linking Methods 0.000 claims abstract description 19
- 238000007605 air drying Methods 0.000 claims abstract description 14
- 238000003756 stirring Methods 0.000 claims abstract description 14
- 238000006243 chemical reaction Methods 0.000 claims abstract description 12
- 239000003431 cross linking reagent Substances 0.000 claims abstract description 11
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims abstract description 9
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000003054 catalyst Substances 0.000 claims abstract description 7
- 239000008367 deionised water Substances 0.000 claims abstract description 7
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 7
- 210000002435 tendon Anatomy 0.000 claims abstract description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000012984 antibiotic solution Substances 0.000 claims abstract description 5
- 230000008439 repair process Effects 0.000 claims description 26
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 claims description 16
- 230000003115 biocidal effect Effects 0.000 claims description 13
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical group ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 10
- 229960003405 ciprofloxacin Drugs 0.000 claims description 8
- GSDSWSVVBLHKDQ-UHFFFAOYSA-N 9-fluoro-3-methyl-10-(4-methylpiperazin-1-yl)-7-oxo-2,3-dihydro-7H-[1,4]oxazino[2,3,4-ij]quinoline-6-carboxylic acid Chemical compound FC1=CC(C(C(C(O)=O)=C2)=O)=C3N2C(C)COC3=C1N1CCN(C)CC1 GSDSWSVVBLHKDQ-UHFFFAOYSA-N 0.000 claims description 7
- 102000002734 Collagen Type VI Human genes 0.000 claims description 7
- 108010043741 Collagen Type VI Proteins 0.000 claims description 7
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 claims description 7
- 229930182566 Gentamicin Natural products 0.000 claims description 7
- 229960001699 ofloxacin Drugs 0.000 claims description 7
- 229960000707 tobramycin Drugs 0.000 claims description 7
- NLVFBUXFDBBNBW-PBSUHMDJSA-N tobramycin Chemical compound N[C@@H]1C[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N NLVFBUXFDBBNBW-PBSUHMDJSA-N 0.000 claims description 7
- 238000000746 purification Methods 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 5
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical group CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 2
- 150000001875 compounds Chemical class 0.000 claims description 2
- 230000003287 optical effect Effects 0.000 abstract description 6
- 238000000465 moulding Methods 0.000 abstract description 2
- 238000005406 washing Methods 0.000 abstract description 2
- 235000015278 beef Nutrition 0.000 abstract 1
- 238000002791 soaking Methods 0.000 abstract 1
- 239000003814 drug Substances 0.000 description 7
- 239000002994 raw material Substances 0.000 description 5
- 238000002054 transplantation Methods 0.000 description 5
- 229940079593 drug Drugs 0.000 description 4
- 241000191967 Staphylococcus aureus Species 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- -1 hydrochloric acid compound Chemical class 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- 238000001356 surgical procedure Methods 0.000 description 3
- 206010061218 Inflammation Diseases 0.000 description 2
- KFSLWBXXFJQRDL-UHFFFAOYSA-N Peracetic acid Chemical compound CC(=O)OO KFSLWBXXFJQRDL-UHFFFAOYSA-N 0.000 description 2
- 210000002919 epithelial cell Anatomy 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 206010023365 keratopathy Diseases 0.000 description 2
- 230000002980 postoperative effect Effects 0.000 description 2
- 201000004569 Blindness Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000425571 Trepanes Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000000845 anti-microbial effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 230000000721 bacterilogical effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 210000000887 face Anatomy 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 206010023332 keratitis Diseases 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000011587 new zealand white rabbit Methods 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 238000013269 sustained drug release Methods 0.000 description 1
- 238000002834 transmittance Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/14—Macromolecular materials
- A61L27/22—Polypeptides or derivatives thereof, e.g. degradation products
- A61L27/24—Collagen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61F—FILTERS IMPLANTABLE INTO BLOOD VESSELS; PROSTHESES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF, TUBULAR STRUCTURES OF THE BODY, e.g. STENTS; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; FOMENTATION; TREATMENT OR PROTECTION OF EYES OR EARS; BANDAGES, DRESSINGS OR ABSORBENT PADS; FIRST-AID KITS
- A61F2/00—Filters implantable into blood vessels; Prostheses, i.e. artificial substitutes or replacements for parts of the body; Appliances for connecting them with the body; Devices providing patency to, or preventing collapsing of, tubular structures of the body, e.g. stents
- A61F2/02—Prostheses implantable into the body
- A61F2/14—Eye parts, e.g. lenses, corneal implants; Implanting instruments specially adapted therefor; Artificial eyes
- A61F2/142—Cornea, e.g. artificial corneae, keratoprostheses or corneal implants for repair of defective corneal tissue
-
- 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
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3687—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by the use of chemical agents in the treatment, e.g. specific enzymes, detergents, capping agents, crosslinkers, anticalcification agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/36—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix
- A61L27/3683—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment
- A61L27/3691—Materials for grafts or prostheses or for coating grafts or prostheses containing ingredients of undetermined constitution or reaction products thereof, e.g. transplant tissue, natural bone, extracellular matrix subjected to a specific treatment prior to implantation, e.g. decellularising, demineralising, grinding, cellular disruption/non-collagenous protein removal, anti-calcification, crosslinking, supercritical fluid extraction, enzyme treatment characterised by physical conditions of the treatment, e.g. applying a compressive force to the composition, pressure cycles, ultrasonic/sonication or microwave treatment, lyophilisation
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
- A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L27/54—Biologically active materials, e.g. therapeutic substances
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- 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
- A61L2300/404—Biocides, antimicrobial agents, antiseptic agents
- A61L2300/406—Antibiotics
-
- 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
- A61L2430/16—Materials or treatment for tissue regeneration for reconstruction of eye parts, e.g. intraocular lens, cornea
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Transplantation (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Oral & Maxillofacial Surgery (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Dermatology (AREA)
- Epidemiology (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Botany (AREA)
- Organic Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- General Chemical & Material Sciences (AREA)
- Vascular Medicine (AREA)
- Toxicology (AREA)
- Zoology (AREA)
- Ophthalmology & Optometry (AREA)
- Biochemistry (AREA)
- Heart & Thoracic Surgery (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Cardiology (AREA)
- Materials For Medical Uses (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
本发明公开了一种抗菌角膜修复材料的制备方法,包括以下步骤:(1)将从牛筋中提取的Ⅰ型胶原纯化,用乙酸或盐酸溶液配制浓度为6.0~10.0mg/ml的胶原溶液;(2)将胶原溶液浇铸到角膜修复材料成型模具中,在室温条件下自然风干成膜;(3)将所得的胶原膜浸泡到抗生素溶液中,搅拌使膜与溶液充分接触;然后向上述溶液中加入交联剂和催化剂,搅拌发生交联反应;(4)将步骤(3)所得的交联膜材料取出,用去离子水冲洗3~5次,然后在室温条件下自然风干,即得到抗菌角膜修复材料。该材料具有较好的力学性能、光学性能和生物相容性,且具有良好的抗菌效果。可用于医疗领域中受损角膜组织的修复和替代。
Description
技术领域
本发明涉及一种抗菌角膜修复材料的制备方法,具体涉及到一种能够在修复受损角膜组织的同时还能降低发生角膜炎症反应的风险的生物材料的制备方法。本发明所得的材料可应用于受损角膜组织的修复和替代。
背景技术
角膜疾病是一种十分常见的眼科病,世界上约有三分之一的失明患者是由于角膜疾病而导致的。角膜移植是治疗角膜盲的一种有效手段,而同种异体的捐赠角膜是目前临床上唯一可行的角膜修复材料。但是由于健康的捐赠角膜的数量远少于角膜移植的需求量,因此人工角膜的研发就显得十分必要。
除了角膜移植材料的匮乏之外,如何避免角膜移植手术后的炎症反应是角膜移植能否成功所面临的另外一个难题。目前常见的方法就是在术后频繁地点药,这种方法不但浪费了大量的药物,同时很难保证药物的用量和加药时间,对患者本身来说又是一件极其麻烦的事情。因此很多组织工程修复材料会通过对药物的包裹或吸附等物理作用来实现药物负载,而针对角膜独特的光学透明性和韧性要求,这类修复材料往往很难保证材料在具有药物缓释效果的同时还具有较好的光学性能和力学性能。因此在保证角膜修复材料所需理化性能和生物学性能的基础上,使之在术后伤口恢复的一段时间里具有抗菌的效果,那将极大地降低角膜移植手术后发生炎症反应的风险。
发明内容
本发明的目的在于克服现有技术的缺点,提供一种新的抗菌角膜修复材料的制备方法。本发明采用高纯度Ι型胶原和眼科手术后常用的抗生素小分子为原料,按照一定的比例,经过模具成膜,抗生素溶液浸泡,搅拌交联,洗涤和风干等工艺,得到一种具有较好的力学性能,光学性能和生物相容性的角膜修复材料。同时,本发明提出的抗菌角膜修复材料还能在一定时间内对导致眼科手术后炎症反应的诸如金黄色葡萄球菌等细菌具有良好的抗菌效果。本材料可用于医疗领域中受损角膜组织的修复和替代,同时还能降低移植手术后细菌感染的风险。
本发明的目的通过以下技术方案实现:
一种抗菌角膜修复材料的制备方法,包括以下步骤:
(1)将从牛筋中提取的Ι型胶原纯化,用乙酸或盐酸溶液配制浓度为6.0~10.0mg/ml的胶原溶液;
(2)将胶原溶液浇铸到角膜修复材料成型模具中,在室温条件下自然风干成膜;
(3)将步骤(2)所得的胶原膜浸泡到浓度为5.0~25.0mg/ml的抗生素溶液中,搅拌使膜与溶液充分接触;然后向上述溶液中加入交联剂和催化剂,搅拌发生交联反应;所述交联剂与催化剂的质量比为4:1,胶原和抗生素的总质量与交联剂的质量比为(5~7):1;
(4)将步骤(3)所得的交联膜材料取出,用去离子水冲洗3~5次,然后在室温条件下自然风干,即得到抗菌角膜修复材料。
优选地,步骤(2)中所述角膜修复材料成型模具具有与角膜组织相似的几何外形。
优选地,步骤(3)所述抗生素的浓度为10.0~15.0mg/ml。
优选地,步骤(3)所述抗生素为妥布霉素、庆大霉素、氧氟沙星或环丙沙星。
优选地,步骤(3)所述交联剂为1-乙基-3(3-二甲基氨丙基)碳化二亚胺(EDC),催化剂为N-羟基丁二酰亚胺(NHS)。
优选地,所述胶原和抗生素的总质量与交联剂的质量比为6:1。
优选地,步骤(3)中所述交联反应的时间为2~6小时。
与现有技术相比,本发明具有如下优点:
(1)本发明制备的角膜修复材料能够在术后伤口恢复的一段时间里具有抗菌性,能够极大地降低角膜移植手术后发生炎症反应的风险。
(2)本发明添加抗生素后制备的角膜修复材料仍然具有与天然角膜组织类似的光学性能、力学性能和生物学性能。
(3)本发明制备的角膜修复材料能够极大地方便患者的术后护理,并降低大量药物所需的费用。
(4)本发明所采用的成型工艺简单,原料成本较低,有利于规模生产。
附图说明
图1是实施例1的Tob-Col膜的样品图。
图2是实施例1的Tob-Col膜与对照组Col膜的透光率曲线图。
图3是实施例1的Tob-Col膜与对照组Col膜的抗张强度图。
图4是金黄色葡萄球菌在Col膜上的生长图。
图5是金黄色葡萄球菌在实施例1的Tob-Col膜上的生长图。
图6是实施例1的Tob-Col膜与对照组Col膜的细胞毒性测试结果图。
图7是人眼角膜上皮细胞在实施例1的Tob-Col膜材料表面的生长图。
图8是实施例1的Tob-Col膜在动物眼表的角膜移植图。
具体实施方式
为了更好的理解本发明,下面结合实施例对本发明做进一步地说明,但本发明要求保护的范围并不局限于此。
实施例1
以妥布霉素(Tobramycin)和胶原(Col)为原料,制备一种妥布霉素-胶原交联膜(Tob-Col)材料。这种Tob-Col膜材料的制备步骤如下:
(1)将从牛筋中提取的Ι型胶原纯化,用盐酸配制浓度为6mg/mL的胶原溶液;
(2)将8mL浓度为6mg/mL的胶原溶液浇铸到角膜修复材料成型模具中,在室温条件下自然风干成膜;
(3)将步骤(2)所得的胶原膜浸泡到浓度为15mg/mL的妥布霉素溶液中,搅拌使胶原膜与妥布霉素溶液充分接触;
(4)在步骤(3)的溶液中加入EDC和NHS(EDC:NHS=4:1),其中胶原和妥布霉素的总质量与EDC的质量比为6:1,搅拌发生交联反应,2小时之后将所得的交联膜材料取出,用去离子水冲洗3次,然后在室温条件下自然风干得到Tob-Col膜。
实施例2
以庆大霉素(Gentamicin)和胶原(Col)为原料,制备一种庆大霉素-胶原交联膜(Gen-Col)材料。这种Gen-Col膜材料的制备步骤如下:
(1)将从牛筋中提取的Ι型胶原纯化,用盐酸配制浓度为8mg/mL的胶原溶液;
(2)将8mL浓度为8mg/mL的胶原溶液浇铸到角膜修复材料成型模具中,在室温条件下自然风干成膜;
(3)将步骤(2)所得的胶原膜浸泡到浓度为15mg/mL的庆大霉素溶液中,搅拌使胶原膜与庆大霉素溶液充分接触;
(4)在步骤(3)的溶液中加入EDC和NHS(EDC:NHS=4:1),其中胶原和庆大霉素的总质量与EDC的质量比为6:1,搅拌3小时使抗生素小分子与胶原发生交联反应;
(5)将步骤(4)所得的交联膜材料取出,用去离子水冲洗3次,然后在室温条件下自然风干得到Gen-Col膜。
实施例3
以环丙沙星(Ciprofloxacin)和胶原(Col)为原料,制备一种环丙沙星-胶原交联膜(Cip-Col)材料。这种Cip-Col膜材料的制备步骤如下:
(1)将从牛筋中提取的Ι型胶原纯化,用乙酸配制浓度为8mg/mL的胶原溶液;
(2)将8mL浓度为8mg/mL的胶原溶液浇铸到角膜修复材料成型模具中,在室温条件下自然风干成膜;
(3)将步骤(2)所得的胶原膜浸泡到浓度为5mg/mL的环丙沙星溶液中,搅拌使胶原膜与环丙沙星溶液充分接触;
(4)在步骤(3)的溶液中加入EDC和NHS(EDC:NHS=4:1),其中胶原和环丙沙星的总质量与EDC的质量比为6:1,搅拌4小时使环丙沙星分子与胶原发生交联反应;
(5)将步骤(4)所得的交联膜材料取出,用去离子水冲洗3次,然后在室温条件下自然风干得到Cip-Col膜。
实施例4
以氧氟沙星(Ofloxacin)和胶原(Col)为原料,制备一种氧氟沙星-胶原交联膜(Ofl-Col)材料。这种Ofl-Col膜材料的制备步骤如下:
(1)将从牛筋中提取的Ι型胶原纯化,用盐酸配制浓度为10mg/mL的胶原溶液;
(2)将8mL浓度为10mg/mL的胶原溶液浇铸到角膜修复材料成型模具中,在室温条件下自然风干成膜;
(3)将步骤(2)所得的胶原膜浸泡到浓度为25mg/mL的氧氟沙星溶液中,搅拌使胶原膜与氧氟沙星溶液充分接触;
(4)在步骤(3)的溶液中加入EDC和NHS(EDC:NHS=4:1),其中胶原和氧氟沙星的总质量与EDC的质量比为6:1,搅拌6小时使抗生素小分子与胶原发生交联反应;
(5)将步骤(4)所得的交联膜材料取出,用去离子水冲洗3次,然后在室温条件下自然风干得到Ofl-Col膜。
通过对实施例1中的材料进行理化性能表征,图1和图2显示该Tob-Col角膜修复材料具有较好的光学性能,图3表明该Tob-Col膜具有合适的力学性能;图4和图5表明该Tob-Col膜相对Col膜而言,具有较好的抗菌性能;图6和图7分别显示了人眼角膜上皮细胞在Tob-Col膜表面的增殖以及细胞的形貌,细胞能够在材料表面稳定增殖,紧密地贴附于材料表面并且保持细胞正常的纺锤形貌。图8是将新西兰大白兔的自身角膜用环钻和刀片切除和剥离后,将Tob-Col膜进行板层移植后的术后照片,可以看到本材料能够耐受眼科手术缝线的缝合,并且能在眼表保持透明。
Claims (6)
1.一种抗菌角膜修复材料的制备方法,其特征在于,包括以下步骤:
(1)将从牛筋中提取的Ι型胶原纯化,用乙酸或盐酸溶液配制浓度为6.0~10.0mg/ml的胶原溶液;
(2)将胶原溶液浇铸到角膜修复材料成型模具中,在室温条件下自然风干成膜;
(3)将步骤(2)所得的胶原膜浸泡到浓度为5.0~25.0mg/ml的抗生素溶液中,搅拌使膜与溶液充分接触;然后向上述溶液中加入交联剂和催化剂,搅拌发生交联反应;所述交联剂与催化剂的质量比为4:1,胶原和抗生素的总质量与交联剂的质量比为(5~7):1;所述抗生素为妥布霉素、庆大霉素、氧氟沙星或环丙沙星,交联剂为1-乙基-3(3-二甲基氨丙基)碳化二亚胺,催化剂为N-羟基丁二酰亚胺;
(4)将步骤(3)所得的交联膜材料取出,用去离子水冲洗3~5次,然后在室温条件下自然风干,即得到抗菌角膜修复材料。
2.根据权利要求1所述的制备方法,其特征在于,步骤(2)中所述角膜修复材料成型模具具有与角膜组织相似的几何外形。
3.根据权利要求1所述的制备方法,其特征在于,步骤(3)所述抗生素的浓度为10.0~15.0mg/ml。
4.根据权利要求1或2或3所述的制备方法,其特征在于,所述胶原和抗生素的总质量与交联剂的质量比为6:1。
5.根据权利要求1或2或3所述的制备方法,其特征在于,步骤(3)中所述交联反应的时间为2~6小时。
6.权利要求1~5任意一项方法制备的抗菌角膜修复材料。
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310181772.5A CN103272268B (zh) | 2013-05-16 | 2013-05-16 | 一种抗菌角膜修复材料及其制备方法 |
US14/891,604 US9585984B2 (en) | 2013-05-16 | 2013-12-20 | Antibacterial cornea repair material and preparation method thereof |
PCT/CN2013/090086 WO2014183445A1 (zh) | 2013-05-16 | 2013-12-20 | 一种抗菌角膜修复材料及其制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201310181772.5A CN103272268B (zh) | 2013-05-16 | 2013-05-16 | 一种抗菌角膜修复材料及其制备方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN103272268A CN103272268A (zh) | 2013-09-04 |
CN103272268B true CN103272268B (zh) | 2015-04-22 |
Family
ID=49054955
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201310181772.5A Active CN103272268B (zh) | 2013-05-16 | 2013-05-16 | 一种抗菌角膜修复材料及其制备方法 |
Country Status (3)
Country | Link |
---|---|
US (1) | US9585984B2 (zh) |
CN (1) | CN103272268B (zh) |
WO (1) | WO2014183445A1 (zh) |
Families Citing this family (17)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103272268B (zh) * | 2013-05-16 | 2015-04-22 | 华南理工大学 | 一种抗菌角膜修复材料及其制备方法 |
CN105037787A (zh) * | 2015-07-21 | 2015-11-11 | 常州大学 | 一种牛磺酸改性胶原基材料及其制备方法 |
CN105148325B (zh) * | 2015-09-18 | 2018-05-15 | 广州市朴道联信生物科技有限公司 | 一种新的角膜组织修复材料及其制备方法 |
CN106913912A (zh) * | 2015-12-28 | 2017-07-04 | 常州亚环环保科技有限公司 | 一种负载氧氟沙星的胶原蛋白膜抗菌修复材料的制备方法 |
CN105854087B (zh) * | 2016-04-07 | 2019-02-19 | 广州市朴道联信生物科技有限公司 | 一种具有基质修复能力的角膜修复材料及其制备方法 |
CN106039403B (zh) * | 2016-07-15 | 2019-12-03 | 广州尤尼智康生物科技有限公司 | 具有生物活性的角膜修复植片的制备方法和角膜修复植片 |
CN113456889B (zh) * | 2016-07-27 | 2022-06-28 | 珐博进(中国)医药技术开发有限公司 | 生物合成角膜 |
RU2635454C1 (ru) * | 2016-11-18 | 2017-11-13 | Федеральное государственное бюджетное научное учреждение "Научно-исследовательский институт глазных болезней" | Способ лечения гнойной язвы роговицы |
CN107261208A (zh) * | 2017-05-19 | 2017-10-20 | 广州市朴道联信生物科技有限公司 | 一种分层次负载药物角膜修复材料的制备方法 |
CN110404110B (zh) * | 2019-08-21 | 2021-10-22 | 华南理工大学 | 一种耐缝合的胶原基角膜再生修复材料及其制备方法 |
CN113456895B (zh) * | 2021-07-20 | 2022-05-06 | 广州市朴道联信生物科技有限公司 | 一种GelMA-胶原双网络抗菌角膜修复材料及其制备方法和应用 |
CN113717431A (zh) * | 2021-08-26 | 2021-11-30 | 常州大学 | 一种负载牛磺酸分子的胶原基支架材料及其制备方法 |
CN113773379B (zh) * | 2021-09-13 | 2023-08-01 | 熹微(苏州)生物医药科技有限公司 | 一种制备聚乙二醇化类胶原蛋白的方法及其应用 |
US11452288B1 (en) | 2022-03-08 | 2022-09-27 | Terry Earl Brady | Innocuous sterilant using hemocyanin and functionalized fullerenes with broad-spectrum intracellular and interstitial microbiocidal and radical scavenging effects for packaged matter, biologics and organics including liquids, gases, tissue, organs, cells, and limbs with copper mediated oxygenation for viability and preservation |
CN114920966B (zh) * | 2022-06-20 | 2023-06-20 | 湖北工业大学 | 一种组织粘接抑菌性水凝胶膜及其制备方法 |
CN115887740B (zh) * | 2022-10-08 | 2024-02-20 | 湖南中腾湘岳生物科技有限公司 | 一种多孔止血粉及其制备方法 |
CN117085182B (zh) * | 2023-08-28 | 2024-04-09 | 山东第一医科大学附属眼科研究所(山东省眼科研究所、山东第一医科大学附属青岛眼科医院) | 一种仿生生物材料的制备方法及其应用 |
Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1461658A (zh) * | 2002-05-31 | 2003-12-17 | 暨南大学 | 生物诱导型活性人工角膜及其使用方法 |
CN101543643A (zh) * | 2009-04-02 | 2009-09-30 | 天津大学 | 具有生物活性的胶原基复合角膜替代物及其制备方法 |
CN101543642A (zh) * | 2009-04-02 | 2009-09-30 | 天津大学 | 胶原基互穿聚合物网络组织工程角膜替代物及其制备方法 |
CN101745145A (zh) * | 2010-01-19 | 2010-06-23 | 温州医学院眼视光研究院 | 治疗性仿生角膜盖及其制备方法 |
CN102552975A (zh) * | 2012-02-28 | 2012-07-11 | 青岛中皓生物工程有限公司 | 一种组织工程人角膜基质载体支架及其制备方法 |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5259998A (en) * | 1991-10-04 | 1993-11-09 | Chiron Ophthalmics, Inc. | Method for casting dissolvable ophthalmic shields in a mold |
US20050163818A1 (en) | 1996-11-05 | 2005-07-28 | Hsing-Wen Sung | Drug-eluting device chemically treated with genipin |
JP2009507110A (ja) * | 2005-09-09 | 2009-02-19 | オタワ ヘルス リサーチ インスティテュート | 相互侵入ネットワーク、およびそれに関連する方法および組成物 |
WO2009146151A2 (en) * | 2008-04-04 | 2009-12-03 | Forsight Labs, Llc | Corneal onlay devices and methods |
CN101480505B (zh) * | 2009-01-23 | 2012-12-19 | 华南理工大学 | 一种表面功能化壳聚糖角膜修复材料的制备方法 |
CN102989038B (zh) * | 2012-11-05 | 2014-07-30 | 天津大学 | 可递送内皮抑制激素的胶原基复合角膜替代物的制备方法和应用 |
CN103272268B (zh) * | 2013-05-16 | 2015-04-22 | 华南理工大学 | 一种抗菌角膜修复材料及其制备方法 |
-
2013
- 2013-05-16 CN CN201310181772.5A patent/CN103272268B/zh active Active
- 2013-12-20 WO PCT/CN2013/090086 patent/WO2014183445A1/zh active Application Filing
- 2013-12-20 US US14/891,604 patent/US9585984B2/en active Active
Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1461658A (zh) * | 2002-05-31 | 2003-12-17 | 暨南大学 | 生物诱导型活性人工角膜及其使用方法 |
CN101543643A (zh) * | 2009-04-02 | 2009-09-30 | 天津大学 | 具有生物活性的胶原基复合角膜替代物及其制备方法 |
CN101543642A (zh) * | 2009-04-02 | 2009-09-30 | 天津大学 | 胶原基互穿聚合物网络组织工程角膜替代物及其制备方法 |
CN101745145A (zh) * | 2010-01-19 | 2010-06-23 | 温州医学院眼视光研究院 | 治疗性仿生角膜盖及其制备方法 |
CN102552975A (zh) * | 2012-02-28 | 2012-07-11 | 青岛中皓生物工程有限公司 | 一种组织工程人角膜基质载体支架及其制备方法 |
Non-Patent Citations (1)
Title |
---|
角膜修复材料的制备及生物相容性研究;龙玉宇;《中国优秀硕士学位论文全文数据库》;20130115;第22-23页 * |
Also Published As
Publication number | Publication date |
---|---|
WO2014183445A1 (zh) | 2014-11-20 |
US20160082151A1 (en) | 2016-03-24 |
CN103272268A (zh) | 2013-09-04 |
US9585984B2 (en) | 2017-03-07 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN103272268B (zh) | 一种抗菌角膜修复材料及其制备方法 | |
Zhou et al. | Genipin-crosslinked polyvinyl alcohol/silk fibroin/nano-hydroxyapatite hydrogel for fabrication of artificial cornea scaffolds—a novel approach to corneal tissue engineering | |
Liu et al. | Marine collagen scaffolds in tissue engineering | |
Wang et al. | Exploring natural silk protein sericin for regenerative medicine: an injectable, photoluminescent, cell-adhesive 3D hydrogel | |
Liang et al. | Fabrication and characters of a corneal endothelial cells scaffold based on chitosan | |
CN106310380B (zh) | 一种纳米纤维化丝素蛋白凝胶及其制备方法 | |
CN103877617B (zh) | 可注射蚕丝素蛋白-海藻酸盐双交联水凝胶及其制备方法和使用方法 | |
CN106310349A (zh) | 一种再生丝素蛋白凝胶膜 | |
Ulag et al. | 3D printed artificial cornea for corneal stromal transplantation | |
WO2016027988A1 (ko) | 초박막 실크피브로인/콜라겐 복합이식체 및 이의 제조방법 | |
Sapru et al. | Non-immunogenic, porous and antibacterial chitosan and Antheraea mylitta silk sericin hydrogels as potential dermal substitute | |
Zhao et al. | Collagen based film with well epithelial and stromal regeneration as corneal repair materials: Improving mechanical property by crosslinking with citric acid | |
CN102580147B (zh) | 人工角膜及其制备方法 | |
Wang et al. | Ductility and porosity of silk fibroin films by blending with glycerol/polyethylene glycol and adjusting the drying temperature | |
CN105037787A (zh) | 一种牛磺酸改性胶原基材料及其制备方法 | |
CN104548201B (zh) | 一种角膜组织修复材料及其制备方法 | |
JP2009516038A (ja) | 架橋されたゼラチン状材料を基剤とする成形体、該成形体の製造方法、及び該成形体の使用 | |
Liang et al. | Tissue-engineered membrane based on chitosan for repair of mechanically damaged corneal epithelium | |
CN101543642B (zh) | 胶原基互穿聚合物网络组织工程角膜替代物及其制备方法 | |
CN101480505B (zh) | 一种表面功能化壳聚糖角膜修复材料的制备方法 | |
Liu et al. | A collagen film with micro-rough surface can promote the corneal epithelization process for corneal repair | |
CN105148325A (zh) | 一种新的角膜组织修复材料及其制备方法 | |
CN1879578A (zh) | 一种具有生物活性的人工角膜的制备方法 | |
US20200282107A1 (en) | Sterile clear concentrated solution of biocompatible collagen, process for the preparation and use thereof | |
WO2016049791A1 (zh) | 一种用于玻璃体替代材料的原位凝胶及其制备方法与应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant |