CN115671405A - 一种关节腔注射凝胶及其制备方法 - Google Patents
一种关节腔注射凝胶及其制备方法 Download PDFInfo
- Publication number
- CN115671405A CN115671405A CN202211389774.9A CN202211389774A CN115671405A CN 115671405 A CN115671405 A CN 115671405A CN 202211389774 A CN202211389774 A CN 202211389774A CN 115671405 A CN115671405 A CN 115671405A
- Authority
- CN
- China
- Prior art keywords
- gel
- carboxymethyl chitosan
- polyethylene glycol
- cavity injection
- active ester
- 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
- 238000002347 injection Methods 0.000 title claims abstract description 28
- 239000007924 injection Substances 0.000 title claims abstract description 28
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 238000001879 gelation Methods 0.000 title description 4
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 claims abstract description 58
- 229920001661 Chitosan Polymers 0.000 claims abstract description 54
- 239000000243 solution Substances 0.000 claims abstract description 35
- 229920001223 polyethylene glycol Polymers 0.000 claims abstract description 30
- 239000002202 Polyethylene glycol Substances 0.000 claims abstract description 28
- 238000004132 cross linking Methods 0.000 claims abstract description 26
- 150000002148 esters Chemical class 0.000 claims abstract description 23
- 238000006243 chemical reaction Methods 0.000 claims abstract description 22
- 238000002156 mixing Methods 0.000 claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 14
- 230000001954 sterilising effect Effects 0.000 claims abstract description 11
- 238000001816 cooling Methods 0.000 claims abstract description 8
- 238000006467 substitution reaction Methods 0.000 claims description 23
- 230000006196 deacetylation Effects 0.000 claims description 22
- 238000003381 deacetylation reaction Methods 0.000 claims description 22
- 239000011259 mixed solution Substances 0.000 claims description 10
- 238000003756 stirring Methods 0.000 claims description 9
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 6
- 230000035484 reaction time Effects 0.000 claims description 5
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 3
- 239000007853 buffer solution Substances 0.000 claims description 3
- 239000003102 growth factor Substances 0.000 claims description 3
- 235000015097 nutrients Nutrition 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 2
- 229940035676 analgesics Drugs 0.000 claims 1
- 239000000730 antalgic agent Substances 0.000 claims 1
- 230000001050 lubricating effect Effects 0.000 abstract description 9
- 239000003431 cross linking reagent Substances 0.000 abstract description 6
- 230000036632 reaction speed Effects 0.000 abstract description 3
- 230000035807 sensation Effects 0.000 abstract description 3
- 239000000499 gel Substances 0.000 description 53
- 239000008055 phosphate buffer solution Substances 0.000 description 26
- 210000001179 synovial fluid Anatomy 0.000 description 13
- 230000008961 swelling Effects 0.000 description 11
- 201000008482 osteoarthritis Diseases 0.000 description 8
- 230000015556 catabolic process Effects 0.000 description 7
- 238000006731 degradation reaction Methods 0.000 description 7
- 238000007789 sealing Methods 0.000 description 6
- 238000010008 shearing Methods 0.000 description 6
- 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 description 5
- 238000007865 diluting Methods 0.000 description 5
- 229920002674 hyaluronan Polymers 0.000 description 5
- 229960003160 hyaluronic acid Drugs 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 239000013589 supplement Substances 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 229920002101 Chitin Polymers 0.000 description 3
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 3
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 3
- 102000016943 Muramidase Human genes 0.000 description 3
- 108010014251 Muramidase Proteins 0.000 description 3
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 3
- 125000003277 amino group Chemical group 0.000 description 3
- 210000000845 cartilage Anatomy 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 230000003412 degenerative effect Effects 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 210000002744 extracellular matrix Anatomy 0.000 description 3
- 229960000274 lysozyme Drugs 0.000 description 3
- 235000010335 lysozyme Nutrition 0.000 description 3
- 239000004325 lysozyme Substances 0.000 description 3
- 239000011664 nicotinic acid Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- 208000006820 Arthralgia Diseases 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 208000008558 Osteophyte Diseases 0.000 description 2
- 208000002193 Pain Diseases 0.000 description 2
- 206010048873 Traumatic arthritis Diseases 0.000 description 2
- 230000008468 bone growth Effects 0.000 description 2
- 210000001612 chondrocyte Anatomy 0.000 description 2
- 230000006835 compression Effects 0.000 description 2
- 238000007906 compression Methods 0.000 description 2
- 201000010934 exostosis Diseases 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 210000001258 synovial membrane Anatomy 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241001260012 Bursa Species 0.000 description 1
- 239000004593 Epoxy Substances 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 206010023230 Joint stiffness Diseases 0.000 description 1
- NNJVILVZKWQKPM-UHFFFAOYSA-N Lidocaine Chemical compound CCN(CC)CC(=O)NC1=C(C)C=CC=C1C NNJVILVZKWQKPM-UHFFFAOYSA-N 0.000 description 1
- 208000001132 Osteoporosis Diseases 0.000 description 1
- 229920002564 Polyethylene Glycol 3500 Polymers 0.000 description 1
- 238000005299 abrasion Methods 0.000 description 1
- 150000001299 aldehydes Chemical class 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 239000007857 degradation product Substances 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000003467 diminishing effect Effects 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000007863 gel particle Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 206010020718 hyperplasia Diseases 0.000 description 1
- 230000036046 immunoreaction Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 210000000281 joint capsule Anatomy 0.000 description 1
- 229960004194 lidocaine Drugs 0.000 description 1
- 210000003041 ligament Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 238000005461 lubrication Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229960005112 moxifloxacin hydrochloride Drugs 0.000 description 1
- IDIIJJHBXUESQI-DFIJPDEKSA-N moxifloxacin hydrochloride Chemical compound Cl.COC1=C(N2C[C@H]3NCCC[C@H]3C2)C(F)=CC(C(C(C(O)=O)=C2)=O)=C1N2C1CC1 IDIIJJHBXUESQI-DFIJPDEKSA-N 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 238000000053 physical method Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- -1 potassium ferricyanide Chemical compound 0.000 description 1
- 229940071643 prefilled syringe Drugs 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 125000006850 spacer group Chemical group 0.000 description 1
- 210000005065 subchondral bone plate Anatomy 0.000 description 1
- 150000005846 sugar alcohols Polymers 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 210000002435 tendon Anatomy 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 239000003932 viscosupplement Substances 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/04—Macromolecular 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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/04—Macromolecular materials
- A61L31/06—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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—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
- A61L31/00—Materials for other surgical articles, e.g. stents, stent-grafts, shunts, surgical drapes, guide wires, materials for adhesion prevention, occluding devices, surgical gloves, tissue fixation devices
- A61L31/14—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
- A61L31/16—Biologically active materials, e.g. therapeutic substances
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Surgery (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Heart & Thoracic Surgery (AREA)
- Vascular Medicine (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明提供了一种关节腔注射凝胶及其制备方法,涉及生物摩擦学领域,具体通过聚乙二醇活性酯与交联用的羧甲基壳聚糖混合发生交联反应,得到凝胶,将凝胶粉碎后与非交联的羧甲基壳聚糖溶液混合,灭菌,冷却后制得关节腔注射凝胶,其中所述的聚乙二醇活性酯与交联用的羧甲基壳聚糖的质量比为1:20‑20:1。此方法制得的关节腔注射凝胶柔韧性更好、无颗粒感,摩擦系数小、润滑性能更好,热稳定性能好,更具有安全性能,并且采用聚乙二醇活性酯做交联剂,反应速度快,所需时间短。
Description
技术领域
本发明属于生物摩擦学领域,具体涉及一种关节腔注射凝胶及其制备方法。
背景技术
骨关节炎(osteoarthritis)是一种非炎症性的退行性关节病,常表现为关节疼痛、僵硬,特别是长时间活动后,该病好发于50岁以上人群,可从20岁开始发病,但大多数无症状,一般不易发现,骨关节炎的患病率随着年龄增长而增加,女性比男性更多见。骨关节炎的主要病理为软骨退行性变性和消失,以及关节边缘韧带附着处和软骨下骨质反应性增生形成骨赘,并由此引起关节疼痛、僵直畸形和功能障碍。
关节包括软骨、滑膜和关节囊,关节软骨的基本成分是软骨细胞和细胞外基质,其中软骨细胞只占1%,而细胞外基质占99%,细胞外基质中约3%为糖胺聚糖。关节滑液是由关节滑囊和腱鞘的滑液膜分泌的,含有类似粘蛋白物质的透明粘质润滑液,有润滑的作用,是人体器官组织的分泌物,起着润滑、滋润器官和排出毒素的作用。若滑液减少,或滑液变得黏稠,就会使代谢产物留于体内,产生各种疾患:如关节滑液随年龄增大而减少,关节缺少润滑剂,关节就会因磨损而出现退行性关节炎、骨刺、骨质疏松等,关节软骨长期缺乏关节滑液还会造成骨关节坏死。
目前,退行性关节炎在发生的早期多采用注射粘弹性补充剂的方法治疗。常用的粘弹性补充剂包括透明质酸和羧甲基壳聚糖(又称羧甲基葡糖胺聚糖,几丁糖),作为传统的粘弹性补充剂,透明质酸已经应用多年,效果良好,但作为新型粘弹性补充剂的羧甲基壳聚糖,作为粘弹性补充剂所产生的治疗效果要优于透明质酸。专利申请CN105709274A公开了一种用于人工关节的可缓释透明质酸的类关节囊温敏凝胶及其制备方法,可获得用于人工关节的可缓释透明质酸的类关节囊温敏凝胶,该凝胶可在体温以下的温度为溶胶,即液体状态,注射进人体后均布在人工关节周围凝胶化,形成类似于关节囊状的隔衬,凝胶化后的多孔网络,形成向人工关节摩擦面输运人工滑液的缓释环境。中国专利CN107349476B公开了一种仿生关节滑液及其制备方法,具体是以羧甲基壳聚糖作为主要原料,分别采用不同交联度的羧甲基壳聚糖与非交联的羧甲基壳聚糖通过物理方法混合制备得到仿生关节滑液,这种方法可形成粘弹性与流动性指标达到正常关节滑液的仿生关节滑液,可在退行性或外伤性关节炎造成关节滑液缺失或粘弹性下降时,作为粘弹性补充剂使用,有效减缓退行性或外伤性关节炎病变的进程,并缓解疼痛。但是其摩擦系数较大,凝胶颗粒感较重,润滑性能较差。
所以,目前缺少一种摩擦系数小、热稳定更好、安全性强且溶胀度小的用于关节腔的注射凝胶。
发明内容
本发明针对现有技术存在的问题,提供了一种关节腔注射凝胶及其制备方法,利用该制备方法得到的关节腔注射凝胶,其摩擦系数小,润滑性能更好,安全性能与热稳定性能更强,形成的凝胶其粘弹性与流动性指标都达到正常关节腔滑液的标准,有效增加骨生长因子并具有止痛消炎的功效。
为实现上述目的,本发明采用的技术方案如下:
本发明提供了一种关节腔注射凝胶的制备方法,通过聚乙二醇活性酯与交联用的羧甲基壳聚糖混合发生交联反应,得到凝胶,将凝胶粉碎后与非交联的羧甲基壳聚糖溶液混合,灭菌,冷却后制得关节腔注射凝胶;
优选地,所述的聚乙二醇活性酯与交联用的羧甲基壳聚糖的质量比为1:20-20:1。
优选地,所述的聚乙二醇活性酯中聚乙二醇的分子量为2000Da-10000Da,2-8臂,其化学结构式如下:
采用聚乙二醇活性酯(PEG-NHS)做交联剂,反应速度快,反应时间短,可减少制备过程中的细菌、内毒素的增长,对比小分子环氧和醛类的交联剂,其毒性更小且分子链长、可控,若分子量太小则分子链不够长;若分子量太大则交联用的活性基团摩尔比太低。
进一步优选地,所述的聚乙二醇活性酯中聚乙二醇的分子量为2000Da-5000Da,2-4臂;
最优选地,所述的聚乙二醇活性酯中聚乙二醇的分子量为3500Da,4臂。
优选地,所述的交联用的羧甲基壳聚糖的脱乙酰度为30-80%,分子量为500-1500kDa,取代度为80%-200%,粘度为300-1000mpa.s(质量分数1%时)。
进一步优选地,所述的交联用的羧甲基壳聚糖的脱乙酰度为40-60%,分子量为600-800kDa,取代度为100%-180%,粘度为300-500mpa.s(质量分数1%时)。
最优选地,所述的交联用的羧甲基壳聚糖的脱乙酰度为50%,分子量为700KDa,取代度为120%,粘度为360mpa.s(质量分数1%时)。
优选地,所述的非交联的羧甲基壳聚糖溶液的脱乙酰度为小于30%,分子量为500-1500kDa,取代度为80%-200%,粘度为300-1000mpa.s(质量分数1%时)。
进一步优选地,所述的非交联的羧甲基壳聚糖溶液的脱乙酰度为10-28%,分子量为600-800kDa,取代度为100%-180%,粘度为300-500mpa.s(质量分数1%时)。
最优选地,所述的非交联的羧甲基壳聚糖溶液的脱乙酰度为25%,分子量为700kDa,取代度为120%,粘度为360mpa.s(质量分数1%时)。
一般来说,所述的羧甲基壳聚糖分子量越大,粘度越大,越接近正常滑液,摩擦越小,降解时间越长。
优选地,所述的交联反应的pH为7-8.5。
优选地,所述的制备方法具体包括以下步骤:
S1、将交联用的羧甲基壳聚糖溶解于磷酸缓冲溶液中,得到混合液1;
S2、将聚乙二醇活性酯加入混合液1中进行交联反应,静置得凝胶;
S3、将凝胶粉碎后添加非交联羧甲基壳聚糖溶液,得混合液2,灭菌后冷却,得关节腔注射凝胶。
优选地,步骤S1所述的磷酸缓冲溶液的浓度为0.2mol/L,pH为7.2。
优选地,步骤S1所述的交联用的羧甲基壳聚糖的质量分数为1%-20%。
优选地,步骤S2所述交联反应的条件为:温度为20-25℃,搅拌速度为60-90rpm,反应时间为10-30min。
进一步优选地,步骤S2所述交联反应的条件为:温度为25℃,搅拌速度为80rpm,反应时间为15min。
优选地,步骤S2所述的静置的时间为2-4h。
进一步优选地,步骤S2所述的静置时间为2h。
优选地,步骤S2中所述的聚乙二醇活性酯需先溶于PBS中。
进一步优选地,所述的PBS的浓度为0.2mol/L,pH为7.2。
优选地,步骤S3所述的凝胶需先加入PBS稀释。
优选地,步骤S3所述的混合液2中还包括骨骼生长因子(skeleton growthfactor,SGF)、消炎止痛药如利多卡因、盐酸莫西沙星、维生素、多肽营养物质。
优选地,步骤S3所述的凝胶与非交联羧甲基壳聚糖溶液的质量比为1:1-9:1。
进一步优选地,步骤S3所述的凝胶与非交联羧甲基壳聚糖溶液的质量比为4:1。
优选地,步骤S3所述的混合液2中非交联羧甲基壳聚糖的浓度为10-50mg/ml。
优选地,步骤S3所述的灭菌条件为:温度115℃-121℃,时间12-45min;所述冷却的温度为18-25℃。
进一步优选地,步骤S3所述的灭菌条件为:温度为121℃,时间为15min;所述的冷却的温度为25℃。
本发明还提供了一种关节腔注射凝胶,采用上述的制备方法制备得到。
本发明高交联度的凝胶,其柔韧性更好,无颗粒感,碳链的增加导致疏水性的增加,使得凝胶溶胀度更低,减少因溶胀体积变大产生的组织压迫;而交联剂中的聚乙二醇使羧甲基壳聚糖和多元醇分子间形成可逆的氢键,增强羧甲基几丁质分子结构的稳定性,减少高温高压对羧甲基几丁质分子链的破坏,提高羧甲基几丁质的灭菌稳定性,使降解时间更长。
非交联的羧甲基壳聚糖溶液的脱乙酰度要求小于30,避免其氨基可能引起的免疫反应,进而影响生物的安全性,还避免和分子链上的羧基发生美拉德反应引起自身交联,破坏产品稳定性,使产品颜色发黄;而制备交联的羧甲基壳聚糖的脱乙酰度要求为30%-80%,因为其氨基需要和交联剂中的活性酯进行交联反应,形成稳定的酰胺键,交联后氨基不能太多;所有羧甲基壳聚糖的羧甲基取代度应在80%-200%最优,在此取代度范围内具有更好的水溶性。
相对于现有技术,本发明具有以下有益效果:
1.采用聚乙二醇活性酯做交联剂,反应速度快,反应时间短,减少污染;
2.制备的高交联度的关节腔注射凝胶,其柔韧性更好,无颗粒感,碳链的增加导致疏水性的增加,使得凝胶溶胀度更低,减少因溶胀体积变大产生的组织压迫;
3.本发明的关节腔注射凝胶摩擦系数小、润滑性能更好;
4.采用溶菌酶对本发明的关节腔注射凝胶进行降解,其降解时间长;
5.本发明制备的关节腔注射凝胶热稳定性能好,更具有安全性能。
具体实施方式
下面结合具体实施例,对本发明作进一步详细的阐述,下述实施例不用于限制本发明,仅用于说明本发明。以下实施例中所使用的实验方法如无特殊说明,实施例中未注明具体条件的实验方法,通常按照常规条件,下述实施例中所使用的材料、试剂等,如无特殊说明,均可从商业途径得到。
实施例1
称取1g聚乙二醇活性酯PEG-NHS(PEH分子量3500,4臂)溶于10ml的0.2mol/LpH7.2的PBS中,称取3g羧甲基壳聚糖(脱乙酰度50%,取代度1.2,分子量700kDa,粘度为360mpa.s)溶于50ml PBS中充分溶解。将上述两种溶液快速搅拌混合均匀,密封置于25℃环境下反应2h,反应后将凝胶剪碎加入PBS稀释至30mg/ml。配置羧甲基壳聚糖(脱乙酰度25%,取代度1.2,分子量700kDa,粘度为360mpa.s)溶液,浓度20mg/ml,pH 7.2。按照4:1质量比将凝胶和溶液通过压缩空气过70目筛网粉碎分散混合,混合15次,将混匀后的凝胶灌装到预灌封注射器中,121℃高温蒸汽灭菌15min,冷却后得样品。
实施例2
称取1.5g聚乙二醇活性酯PEG-NHS(PEH分子量5000,4臂)溶于10ml的0.2mol/LpH7.2的PBS中,称取3g羧甲基壳聚糖(脱乙酰度45%,取代度1,分子量500kDa,粘度为300mpa.s)溶于50ml PBS中充分溶解。将上述两种溶液快速搅拌混合均匀,密封置于25℃环境下反应2h,反应后将凝胶剪碎加入PBS稀释至30mg/ml。配置羧甲基壳聚糖(脱乙酰度30%,取代度0.85,分子量500kDa,粘度为300mpa.s)溶液,浓度20mg/ml,pH 7.2。按照3:1凝胶和溶液通过压缩空气过70目筛网粉碎分散混合,混合15次,将混匀后的凝胶灌装到预灌封注射器中,121℃高温蒸汽灭菌15min,冷却后得样品。
实施例3
称取3g聚乙二醇活性酯PEG-NHS(PEH分子量2000,2臂)溶于10ml的0.2mol/LpH7.2的PBS中,称取3g羧甲基壳聚糖(脱乙酰度55%,取代度1,分子量600kDa,粘度为330mpa.s)溶于50ml PBS中充分溶解。将上述两种溶液快速搅拌混合均匀,密封置于25℃环境下反应2h,反应后将凝胶剪碎加入PBS稀释至30mg/ml。配置羧甲基壳聚糖(脱乙酰度25%,取代度0.85,分子量600kDa,粘度为330mpa.s)溶液,浓度20mg/ml,pH7.2。按照2:1凝胶和溶液通过压缩空气过70目筛网粉碎分散混合,混合15次,将混匀后的凝胶灌装到预灌封注射器中,121℃高温蒸汽灭菌15min,冷却后得样品。
对比例1
称取1g聚乙二醇活性酯PEG-NHS(PEG分子量1500,4臂)溶于10ml的0.2mol/LpH7.2的PBS中,称取3g羧甲基壳聚糖(脱乙酰度50%,取代度1.2,分子量700kDa,粘度为360mpa.s)溶于50ml PBS中充分溶解。将上述两种溶液快速搅拌混合均匀,密封置于25℃环境下反应2h,反应后将凝胶剪碎加入PBS稀释至30mg/ml。配置羧甲基壳聚糖(脱乙酰度25%,取代度1.2,分子量700kDa,粘度为360mpa.s)溶液,浓度20mg/ml,pH 7.2。按照4:1质量比将凝胶和溶液通过压缩空气过70目筛网粉碎分散混合,混合15次,将混匀后的凝胶灌装到预灌封注射器中,121℃高温蒸汽灭菌15min,冷却后得样品。
对比例2
称取1g聚乙二醇活性酯PEG-NHS(PEG分子量15000,4臂)溶于10ml的0.2mol/LpH7.2的PBS中,称取3g羧甲基壳聚糖(脱乙酰度50%,取代度1.2,分子量700kDa,粘度为360mpa.s)溶于50ml PBS中充分溶解。将上述两种溶液快速搅拌混合均匀,密封置于25℃环境下反应2h,反应后将凝胶剪碎加入PBS稀释至30mg/ml。配置羧甲基壳聚糖(脱乙酰度25%,取代度1.2,分子量700kDa,粘度为360mpa.s)溶液,浓度20mg/ml,pH 7.2。按照4:1质量比将凝胶和溶液通过压缩空气过70目筛网粉碎分散混合,混合15次,将混匀后的凝胶灌装到预灌封注射器中,121℃高温蒸汽灭菌15min,冷却后得样品。
对比例3
称取0.1g聚乙二醇活性酯PEG-NHS(PEG分子量3500,4臂)溶于10ml的0.2mol/LpH7.2的PBS中,称取3g羧甲基壳聚糖(脱乙酰度50%,取代度1.2,分子量700kDa,粘度为360mpa.s)溶于50ml PBS中充分溶解。将上述两种溶液快速搅拌混合均匀,密封置于25℃环境下反应2h,反应后将凝胶剪碎加入PBS稀释至30mg/ml。配置羧甲基壳聚糖(脱乙酰度25%,取代度1.2,分子量700kDa,粘度为360mpa.s)溶液,浓度20mg/ml,pH 7.2。按照4:1质量比将凝胶和溶液通过压缩空气过70目筛网粉碎分散混合,混合15次,将混匀后的凝胶灌装到预灌封注射器中,121℃高温蒸汽灭菌15min,冷却后得样品。
测试例
粘弹性检测
参照专利申请CN107349476A中第0031-0032段的方法,检测关节腔注射凝胶的流变学数据。结果显示,各样品弹性模量与粘性模量指标均与正常滑液的指标相近,具体结果如表1所示。
表1粘弹性模量数据
润滑能力检测
根据专利CN114634585A第0331段的方法测量摩擦系数,检测凝胶的润滑能力。结果显示,本发明的关节腔注射凝胶的摩擦系数更低,润滑性能更好,与市售产品Syvnisc具有更低的摩擦系数,具体结果如下表所示。
表2润滑能力数据
体外降解能力检测
根据专利CN107179381A实施例4中所述的六重交联羧甲基壳聚糖凝胶的溶菌酶降解的方法,采用溶菌酶对混合凝胶进行降解,并采用GB/T5009.7-2016《食品安全国家标准食品中还原糖的测定》中第三法铁氰化钾还原糖测定法对降解产物进行测定。具体结果如下表所示,本发明的关节腔注射凝胶完全降解时间更长,体外降解能力强。
表3体外降解时间数据
稳定性检测
将混匀后的凝胶灌装到预灌封注射器中,121℃高温蒸汽灭菌15min,将灭菌后的样品和灭菌前的样品进行外观对比和弹性模量检测,发现本发明中的关节腔注射凝胶稳定性更佳,具体结果如下表所示。
表4稳定性数据
安全性检测
根据GB/T16886.5-2017体外细胞毒性试验方法对样品进行检测。结果显示,本发明的关节腔注射凝胶细胞存活率更高,更具有安全性,具体数据如下表所示。
表5安全性数据
溶胀度检测
根据CN110643057A中第0154-0155段的方法检测凝胶的溶胀度,按下式计算凝胶溶胀率。溶胀率=(溶胀后样品质量-取样量)×100%/取样量。结果显示,本发明的关节腔注射凝胶溶胀度更低,具体结果如下表所示。
表6溶胀度数据
最后应当说明的是,以上内容仅用以说明本发明的技术方案,而非对本发明保护范围的限制,本领域的普通技术人员对本发明的技术方案进行的简单修改或者等同替换,均不脱离本发明技术方案的实质和范围。
Claims (10)
1.一种关节腔注射凝胶的制备方法,其特征在于:通过聚乙二醇活性酯与交联用的羧甲基壳聚糖混合发生交联反应,得到凝胶,将凝胶粉碎后与非交联的羧甲基壳聚糖溶液混合,灭菌,冷却后制得关节腔注射凝胶;其中,所述的聚乙二醇活性酯与交联用的羧甲基壳聚糖的质量比为1:20-20:1。
2.根据权利要求1所述的制备方法,其特征在于:所述的聚乙二醇活性酯包括聚乙二醇,所述聚乙二醇的分子量为2000Da-10000Da,2-8臂。
3.根据权利要求1所述的制备方法,其特征在于:所述的交联用的羧甲基壳聚糖的脱乙酰度为30%-80%,分子量500-1500kDa,取代度为80%-200%,粘度300-1000mpa.s。
4.根据权利要求1所述的制备方法,其特征在于:所述的非交联的羧甲基壳聚糖溶液的脱乙酰度小于30%,分子量500-1500kDa,取代度为80%-200%,粘度300-1000mpa.s。
5.根据权利要求1所述的制备方法,其特征在于:具体包括以下步骤:
S1、将交联用的羧甲基壳聚糖溶解于磷酸缓冲溶液中,得到混合液1;
S2、将聚乙二醇活性酯加入混合液1中进行交联反应,静置得凝胶;
S3、将凝胶粉碎后添加非交联羧甲基壳聚糖溶液,得混合液2,灭菌后冷却,得关节腔注射凝胶。
6.根据权利要求5所述的制备方法,其特征在于:步骤S1所述的混合液1中的交联用的羧甲基壳聚糖的质量分数为1-20%。
7.根据权利要求5所述的制备方法,其特征在于:步骤S2所述交联反应的条件为:温度为20-25℃,pH为7-8.5,搅拌速度为60-90rpm,反应时间为10-30min。
8.根据权利要求5所述的制备方法,其特征在于:步骤S3所述的凝胶与非交联羧甲基壳聚糖溶液的质量比为1:1-9:1。
9.根据权利要求5所述的制备方法,其特征在于:步骤S3所述的混合液2中的非交联羧甲基壳聚糖的浓度为10-50mg/ml。
10.一种关节腔注射凝胶,其特征在于:采用权利要求1-9任一项所述的制备方法制备得到,
所述的关节腔注射凝胶还包括生长因子、止痛药和营养物质。
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202211389774.9A CN115671405B (zh) | 2022-11-08 | 2022-11-08 | 一种关节腔注射凝胶及其制备方法 |
PCT/CN2023/126089 WO2024099063A1 (zh) | 2022-11-08 | 2023-10-24 | 一种关节腔注射凝胶及其制备方法 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202211389774.9A CN115671405B (zh) | 2022-11-08 | 2022-11-08 | 一种关节腔注射凝胶及其制备方法 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN115671405A true CN115671405A (zh) | 2023-02-03 |
CN115671405B CN115671405B (zh) | 2023-11-10 |
Family
ID=85050654
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202211389774.9A Active CN115671405B (zh) | 2022-11-08 | 2022-11-08 | 一种关节腔注射凝胶及其制备方法 |
Country Status (2)
Country | Link |
---|---|
CN (1) | CN115671405B (zh) |
WO (1) | WO2024099063A1 (zh) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2024099063A1 (zh) * | 2022-11-08 | 2024-05-16 | 北京大清生物技术股份有限公司 | 一种关节腔注射凝胶及其制备方法 |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090117070A1 (en) * | 2004-06-23 | 2009-05-07 | Angiotech Pharmaceuticals (Us), Inc. | Methods and Crosslinked Polymer Compositions for Cartilage Repair |
CN107349476A (zh) * | 2017-06-13 | 2017-11-17 | 爱美客技术发展股份有限公司 | 仿生关节滑液及其制备方法 |
US20180214561A1 (en) * | 2015-06-23 | 2018-08-02 | Xiamen Sinopeg Biotech Co., Ltd. | Eight-arm polyethylene glycol derivative, production method therefor, and modifiedbio-relatedsubstancethereof |
CN112675355A (zh) * | 2020-12-16 | 2021-04-20 | 北京大清生物技术股份有限公司 | 一种可降解医用水凝胶材料及其制备方法与应用 |
US20210161672A1 (en) * | 2018-06-11 | 2021-06-03 | Histogenics Corporation | Scaffold with adhesive for articular cartilage repair |
CN113797385A (zh) * | 2021-08-18 | 2021-12-17 | 山东大学 | 一种壳聚糖/聚乙二醇水凝胶及其制备方法与应用 |
CN114732943A (zh) * | 2022-04-19 | 2022-07-12 | 中国科学院合肥物质科学研究院 | 基于壳聚糖-活性酯凝胶的抗菌材料及其制备方法与应用 |
CN115252875A (zh) * | 2021-04-29 | 2022-11-01 | 浙江大学 | 一种医用组织粘合胶及其制备方法 |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US8754564B2 (en) * | 2010-05-27 | 2014-06-17 | Covidien Lp | Hydrogel implants with varying degrees of crosslinking |
US20140170224A1 (en) * | 2012-12-14 | 2014-06-19 | Agency For Science, Technology And Research | Gelatin-based microgels |
US10173357B2 (en) * | 2013-09-26 | 2019-01-08 | Northwestern University | Poly(ethylene glycol) cross-linking of soft materials to tailor viscoelastic properties for bioprinting |
CN111939324B (zh) * | 2020-08-14 | 2022-06-17 | 深圳市人民医院 | 一种天然多糖基可注射原位成型水凝胶及其制备方法和应用 |
CN115671405B (zh) * | 2022-11-08 | 2023-11-10 | 北京大清生物技术股份有限公司 | 一种关节腔注射凝胶及其制备方法 |
-
2022
- 2022-11-08 CN CN202211389774.9A patent/CN115671405B/zh active Active
-
2023
- 2023-10-24 WO PCT/CN2023/126089 patent/WO2024099063A1/zh unknown
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20090117070A1 (en) * | 2004-06-23 | 2009-05-07 | Angiotech Pharmaceuticals (Us), Inc. | Methods and Crosslinked Polymer Compositions for Cartilage Repair |
US20180214561A1 (en) * | 2015-06-23 | 2018-08-02 | Xiamen Sinopeg Biotech Co., Ltd. | Eight-arm polyethylene glycol derivative, production method therefor, and modifiedbio-relatedsubstancethereof |
CN107349476A (zh) * | 2017-06-13 | 2017-11-17 | 爱美客技术发展股份有限公司 | 仿生关节滑液及其制备方法 |
US20210161672A1 (en) * | 2018-06-11 | 2021-06-03 | Histogenics Corporation | Scaffold with adhesive for articular cartilage repair |
CN112675355A (zh) * | 2020-12-16 | 2021-04-20 | 北京大清生物技术股份有限公司 | 一种可降解医用水凝胶材料及其制备方法与应用 |
CN115252875A (zh) * | 2021-04-29 | 2022-11-01 | 浙江大学 | 一种医用组织粘合胶及其制备方法 |
CN113797385A (zh) * | 2021-08-18 | 2021-12-17 | 山东大学 | 一种壳聚糖/聚乙二醇水凝胶及其制备方法与应用 |
CN114732943A (zh) * | 2022-04-19 | 2022-07-12 | 中国科学院合肥物质科学研究院 | 基于壳聚糖-活性酯凝胶的抗菌材料及其制备方法与应用 |
Non-Patent Citations (2)
Title |
---|
CHUANXU YANG等: "Impact of PEG Chain Length on the Physical Properties and Bioactivity of PEGylated Chitosan/siRNA Nanoparticles in Vitro and in Vivo", ACS APPLIED MATERIALS & INTERFACES, vol. 9, pages 12203 * |
王婷 等: "聚乙二醇化壳聚糖的制备及其应用研究进展", 化学通报, vol. 83, no. 6, pages 536 - 545 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2024099063A1 (zh) * | 2022-11-08 | 2024-05-16 | 北京大清生物技术股份有限公司 | 一种关节腔注射凝胶及其制备方法 |
Also Published As
Publication number | Publication date |
---|---|
WO2024099063A1 (zh) | 2024-05-16 |
CN115671405B (zh) | 2023-11-10 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US8524213B2 (en) | Polymeric materials, their preparation and use | |
EP2783702B1 (en) | Water insoluble gel composition and method for preparing same | |
EP1753787B1 (en) | Method of covalently linking hyaluronan and chitosan | |
EP1701981B1 (en) | Cohesive gels from cross-linked hyaluronan and/or hylan,their preparation and use | |
AU2011334627C1 (en) | Glucan compositions | |
US20140343013A1 (en) | Stabalized Glycosaminoglycan Preparations and Related Methods | |
EP2614828A2 (en) | Low-modification biocompatible high polymer sulfhydryl-modified derivatives, cross-linked material thereof, and uses of said material | |
CN101244290A (zh) | 一种用于组织填充的交联透明质酸微粒凝胶的制备方法 | |
CN104086788A (zh) | 一种注射用修饰透明质酸钠凝胶 | |
CN103333349A (zh) | 一种注射用透明质酸-胶原蛋白复合水凝胶及其制备方法 | |
CN113429589B (zh) | 甘草酸基pH敏感型缓释水凝胶材料及其制备方法与应用 | |
CN115429935B (zh) | 一种可注射性的交联硫酸软骨素水凝胶及其制备方法 | |
CN115671405B (zh) | 一种关节腔注射凝胶及其制备方法 | |
CN114502599B (zh) | 一种超支化聚甘油多缩水甘油醚及其作为多糖交联剂的用途 | |
CN1593385A (zh) | 可注射温敏配合物凝胶及其制备方法 | |
CN115554462A (zh) | 一种抗溶胀的可注射水凝胶粘接剂及其制备方法 | |
Zheng et al. | Fabrication of phenylalanine amidated pectin using ultra-low temperature enzymatic method and its hydrogel properties in drug sustained release application | |
CN107029281A (zh) | 一种可吸收止血材料的制备方法 | |
CN112516075B (zh) | 一种载有泼尼松的透明质酸-壳聚糖温敏水凝胶及其制备方法 | |
CN104231285A (zh) | 透明质酸衍生物凝胶及其制备方法 | |
CN107349476B (zh) | 仿生关节滑液及其制备方法 | |
CN110655662B (zh) | 一种橄榄苦苷交联制备透明质酸钠凝胶的方法及透明质酸钠凝胶 | |
JPH09327507A (ja) | 創傷包帯剤として使用するための固体の多糖材料 | |
EP3231455A1 (en) | Biocompatible composition and method for preparing same | |
CN107362352B (zh) | 一种蛋白或多肽组合物及其制备方法和用途 |
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 |