动态压力联合胰岛素样生长因子1基因转染对壳聚糖/明胶支架中兔脂肪间充质干细胞成软骨分化的影响  

Effect of dynamic compression combined with insulin-like growth factor-1 gene transfection on chondrogenic differentiation of rabbit adipose-derived mesenchymal stem cells in chitosan/gelatin scaffolds

作　　者：张传辉 李建军[2] 杨军[2] Zhang Chuanhui;Li Jianjun;Yang Jun(Department of Orthopeadic Surgery,Chaoyang City Center Hospital,Chaoyang 122000,Liaoning Province,China;Department of Orthopedic Trauma,Shengjing Hospital of China Medical University,Shenyang 110004,Liaoning Province,China)

机构地区：[1]朝阳市中心医院骨外科,辽宁省朝阳市122000 [2]中国医科大学附属盛京医院创伤骨科,辽宁省沈阳市110004

出　　处：《中国组织工程研究》2021年第28期4485-4491,共7页Chinese Journal of Tissue Engineering Research

基　　金：辽宁省自然科学基金资助项目(20102264),项目负责人:李建军。

摘　　要：背景:应用生物反应器体外模拟人体内生物力学环境,联合应用基因转染技术体外构建组织工程软骨是目前组织工程研究领域的新思路。对于机械应力促进脂肪间充质干细胞成软骨分化的作用机制,目前国内外尚无定论。目的:研究动态压力与胰岛素样生长因子1(insulin-like growth factor-1,IGF-1)基因转染对诱导负载于壳聚糖/明胶复合支架上的兔脂肪间充质干细胞成软骨分化的交互作用。方法:将壳聚糖/明胶复合支架材料置于6孔培养板中,分4组干预:A组滴加脂肪间充质干细胞悬液,孵育4 h后加入完全培养基培养9 d;B组滴加IGF-1转染的脂肪间充质干细胞悬液,孵育4 h后加入完全培养基培养9 d;C组滴加脂肪间充质干细胞悬液,孵育4 h后加入完全培养基培养2 d,随后进行动态压力培养(2%形变,频率1 Hz),加压20 min,间歇20 min,每天6个循环周期,继续培养7 d;D组滴加IGF-1转染的脂肪间充质干细胞悬液,孵育4 h后加入完全培养基培养2 d,随后进行动态压力培养,继续培养7 d。培养结束后,检测细胞-支架复合物中细胞的增殖速度、糖胺聚糖含量、钙离子含量及软骨相关基因表达(IGF-1、Sox-9、Ⅱ型胶原、蛋白聚糖和X型胶原)。结果与结论:①各组细胞均具有较好的增殖能力,其中增殖速度由快至慢依次为D组、B组、C组、A组;②B、C、D组的糖胺聚糖、钙离子含量高于A组(P<0.01),D组高于B、C组(P<0.01),B组高于C组(P<0.01);③4组间X型胶原mRNA表达无差异(P﹥0.05);B、C、D组的IGF-1、Sox-9、Ⅱ型胶原、蛋白聚糖mRNA表达高于A组(P<0.01),D组高于B、C组(P<0.01),B组高于C组(P<0.01);④结果表明,动态压力联合IGF-1基因转染协同诱导壳聚糖/明胶复合支架上脂肪间充质干细胞的成软骨分化。BACKGROUND:It is a new idea that the application of bioreactor to simulate the biomechanical environment of human body, and the combination with gene transfection technology to construct tissue-engineered cartilage in vitro in the field of tissue engineering research.At present, there is no conclusion about the action mechanism of mechanical stress promoting the chondrogenic differentiation of adipose-derived mesenchymal stem cells at home and abroad.OBJECTIVE:To investigate the comparative and interactive effects of dynamic compression and insulin-like growth factor-1 on the chondrogenesis of rabbit adipose-derived mesenchymal stem cells in chitosan/gelatin scaffolds.METHODS:Chitosan/gelatin composite scaffolds were placed in a six-well culture plate and divided into four groups.In group A, rabbit adipose-derived mesenchymal stem cells were added into the complete medium for 9 days after incubation for 4 hours.In group B, rabbit adipose-derived mesenchymal stem cells transfected with insulin-like growth factor-1 were incubated for 4 hours and then cultured in complete medium for 9 days.In group C, rabbit adiposederived mesenchymal stem cells with insulin-like growth factor-1 had been cultured in chitosan/gelatin scaffolds for 2 days before dynamic compression after incubation for 4 hours.The cells/scaffold constructs were subjected to cyclic compression with 2% strain and 1 Hz, pressure for 20 minutes, interval of 20 minutes, 6 cycles per day for 7 consecutive days.In group D, adipose-derived mesenchymal stem cells transfected with insulin-like growth factor-1 were incubated for 4 hours in chitosan/gelatin scaffolds and then cultured in complete medium for 2 days, followed by dynamic pressure culture for 7 days.After the trial intervention, the cell proliferation rate, total glycosaminoglycan content, calcium content and cartilage related gene expression(insulin-like growth factor 1, Sox-9, type II collagen, aggrecan and type X collagen) were quantified.RESULTS AND CONCLUSION:(1) All the cells in each group had

关 键 词：软骨 因子 动态压力 干细胞 胰岛素样生长因子1 组织工程 软骨细胞 钙离子 钙离子通道 

分 类 号：R459.9[医药卫生—治疗学] R329.25[医药卫生—临床医学]