载神经生长因子软骨及软骨下骨双层仿生支架修复兔软骨缺损  

Repair of rabbit cartilage defects with double-layer bionic scaffold loaded with nerve growth factor cartilage and subchondral bone

作　　者：周杰 叶鹏 张天喜 李兴屿 李沙沙 喻安永 邓江[2] Zhou Jie;Ye Peng;Zhang Tianxi;Li Xingyu;Li Shasha;Yu Anyong;Deng Jiang(Emergency Department of Affiliated Hospital of Zunyi Medical University,Zunyi 563003,Guizhou Province,China;Department of Orthopedics,Zunyi First People’s Hospital,Zunyi 563003,Guizhou Province,China)

机构地区：[1]遵义医科大学附属医院急诊科,贵州省遵义市563003 [2]遵义市第一人民医院骨科,贵州省遵义市563003

出　　处：《中国组织工程研究》2023年第34期5421-5429,共9页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(H0606)地区科学基金,基金名称:SF/CS/nHA仿生支架结合骨软骨镶嵌移植术和PRP对大面积骨软骨缺损修复的研究,项目负责人:邓江;贵州省科技计划项目(黔科合支撑[2021]074号),基金名称:基于HIF-1a信号通路NGF-BMSC/ColⅡ/ColⅠ-SF-CS-nHA种植体软骨缺损修复作用机制研究,项目负责人:叶鹏;遵义市科技联合基金[遵市科合HZ字(2020)247号],基金名称:NGF/Co1Ⅱ-CS/Co1Ⅰ-SF-CS-nHA双相支架基于BMP-2信号通路修复软骨缺损的研究,项目负责人:叶鹏;遵义医科大学博士启动基金[院字(2018)02号],基金名称:BMP-2/BMSCs联合抗生素支架修复骨缺损的实验研究,项目负责人:叶鹏。

摘　　要：背景:大面积骨软骨缺损修复效果差,双层或多层支架由于更加符合软骨解剖、更具仿生性,成为一种新的治疗策略。目的:探讨神经生长因子/Ⅱ型胶原蛋白-丝素蛋白-壳聚糖/Ⅰ型胶原蛋白-丝素蛋白-壳聚糖-纳米羟基磷灰石软骨及软骨下骨双层仿生支架修复软骨缺损的效果。方法:采用冷冻干燥、乳化-溶剂挥发法制备不同比例的Ⅱ型胶原蛋白-丝素蛋白-壳聚糖/Ⅰ型胶原蛋白-丝素蛋白-壳聚糖-纳米羟基磷灰石双层支架,检测支架的理化性能,筛选较适宜的各成分比例支架用于动物实验。将双层支架浸泡于神经生长因子缓释微球溶液中,制备载神经生长因子的双层支架。取30只新西兰大白兔,按照随机数字表法分为3组,每组10只,均建立骨软骨缺损模型,空白对照组不进行任何治疗,对照组植入未载神经生长因子的双层支架,实验组植入载神经生长因子的双层支架,术后4,8,12周,分别进行软骨修复大体观察、软骨组织形态观察。结果与结论:(1)通过孔隙率、吸水膨胀率、热水溶失率等指标的相关结果,得出:上层支架中Ⅱ型胶原蛋白:丝素蛋白:壳聚糖的质量比为1:1:1、下层支架中Ⅰ型胶原蛋白:丝素蛋白:壳聚糖:纳米羟基磷灰石质量比为1:1:1:1时,双层支架具有良好的理化性能,适宜用于动物实验。(2)神经生长因子缓释微球的平均粒径为(25.87±6.54)μm,载药量为0.516μg/mg,包封率为40.5%。(3)动物实验中,大体观察结果显示,实验组软骨修复快于对照组、空白对照组,且修复效果最好。苏木精-伊红染色与阿利新蓝染色显示,至术后12周时,空白对照组缺损区可见纤维组织及少量软骨陷窝结构,对照组缺损区修复组织大部分为纤维软骨,软骨下骨未完全重建,骨软骨层分界不清晰,实验组软骨层与软骨下骨层修复比较完整、分界明显,可见潮线。免疫组化染色显示,至术后12周时,实验组修�BACKGROUND:The outcome of large osteochondral defects is poor.Double-layer or multi-layer scaffolds have become a new treatment strategy because they are more consistent with cartilage anatomy and mimicry.OBJECTIVE:To investigate the repair effect of nerve growth factor/type II collagen-silk fibroin-chitosan/type I collagen-silk fibroin-chitosan-nano hydroxyapatite cartilage and subchondral bone bionic scaffold in the repair of cartilage defects.METHODS:Double-layer scaffolds loaded with type II collagen-silk fibroin-chitosan/type I collagen-silk fibroin-chitosan-nano hydroxyapatite were prepared by freeze-drying,emulsification,and solvent evaporation.The physical and chemical properties of the scaffolds were tested and the appropriate proportion of scaffold components was selected for animal experiments.A double-layer scaffold containing nerve growth factor was prepared by immersion in nerve growth factor sustained-release microsphere solution.Thirty New Zealand white rabbits were randomly divided into three groups(n=10).An osteochondral defect model was established.The blank control group did not receive any treatment.The control group was implanted with a double-layer scaffold without nerve growth factor.The experimental group was implanted with a double-layer scaffold loaded with nerve growth factor.The gross observation of cartilage repair and the morphological observation of cartilage tissue were performed 4,8 and 12 weeks after operation.RESULTS AND CONCLUSION:(1)The relevant results of porosity,water absorption expansion rate,and hot water dissolution rate concluded that when the mass ratio of type II collagen:silk fibroin:chitosan in the upper scaffold was 1:1:1,and the mass ratio of type I collagen:silk fibroin:chitosan:nano-hydroxyapatite in the lower scaffold was 1:1:1:1.The double-layer scaffold had good physical and chemical properties and was suitable for animal experiments.(2)The average particle size of the sustained release of nerve growth factor microspheres was(25.87±6.54)μm,the drug loading

关 键 词：双层支架 神经生长因子 缓释微球 软骨缺损 组织工程学 胶原蛋白:生物相容性 

分 类 号：R459.9[医药卫生—治疗学] R318.08[医药卫生—临床医学] R-331