脂肪干细胞-丝素/壳聚糖支架复合物在外泌体诱导下的体外成骨效应  

Osteogenic Effect of Exosomes on Adipose-Derived Stem Cells and Silk Fibroin/Chitosan Scaffold Complexes in vitro

作　　者：魏松乔 郭澍[1] 佟爽[1] 朱梦茹 张华[1] 陈义庆 崔梦莹 WEI Songqiao;GUO Shu;TONG Shuang;ZHU Mengru;ZHANG Hua;CHEN Yiqing;CUI Mengying(Department of Plastic Surgery,The First Hospital,China Medical University,Shenyang 110001,China)

机构地区：[1]中国医科大学附属第一医院整形外科

出　　处：《中国医科大学学报》2019年第10期891-895,共5页Journal of China Medical University

基　　金：国家自然科学基金(51272286)

摘　　要：目的探究成骨分化脂肪干细胞(ADSCs)来源的外泌体对ADSCs-丝素/壳聚糖三维支架复合物(ADSCs-SF/CS)的成骨效应。方法体外提取、培养和鉴定ADSCs,采用冷冻干燥法制备三维支架。将ADSCs诱导为成骨细胞,超速离心提取外泌体。将ADSCs接种于支架上,扫描电镜观察。将细胞支架复合体分为普通培养液组(阴性对照组)、成骨诱导液组(阳性对照组)、普通培养液加外泌体组(实验组a)、成骨诱导液加外泌体组(实验组b),连续培养14 d后,采用Western blotting检测成骨相关蛋白Runx2的表达量以证实ADSCs成骨分化,并通过CCK-8法及碱性磷酸酶(ALP)活性检测探究外泌体对ADSCs-SF/CS复合物中ADSCs体外增殖及成骨分化效应的作用。结果CCK-8结果显示,实验组a与阴性对照组相比,加入外泌体有利于ADSCs的增殖,Runx2及ALP活性结果显示,实验组b与阳性对照组相比,加入外泌体可使ADSCs-SF/CS复合物中的ADSCs成骨效应显著增强。结论成骨诱导后,ADSCs来源的外泌体可诱导附着于SF/CS上的ADSCs向成骨细胞分化,并且对ADSCs的成骨具有促进效应。Objective To investigate the osteogenic effect of exosomes on adipose-derived stem cells(ADSCs)and silk fibroin/chitosan(SF/CS)3D scaffold complexes in vitro.Methods ADSCs were extracted,cultivated,and identified.SF/CS scaffolds were prepared using a freeze-drying method.ADSCs were induced to differentiate into osteoblasts,and exosomes were extracted by ultracentrifugation.ADSCs were seeded onto the SF/CS scaffolds and observed using a scanning electron microscope.The cell-scaffold complexes were divided into four groups for testing:the common culture group(negative control group),osteogenesis-induced liquid group(positive control group),common culture plus exosomes group(experimental group a),and osteogenesis-induced liquid plus exosomes group(experimental group b).After fostering the complexes for 14 days,the osteogenic effect of exosomes on ADSCs was evaluated by measuring the expression of the osteogenesis-related protein Runx2 and the activity of alkaline phosphatase(ALP).Results Cell counting assays comparing experimental group a and the negative control group revealed that adding exosomes to the culture media was beneficial to the proliferation of ADSCs.Comparison of Runx2 expression and ALP activity between experimental group b and the positive control group indicated that the addition of exosomes significantly enhanced the osteogenic effect of ADSCs in the cell-scaffold complexes.Conclusion Exosomes derived from ADSCs induce the differentiation of ADSCs into osteoblasts and promote the osteogenesis of ADSCs in vitro.

关 键 词：脂肪干细胞 丝素/壳聚糖三维支架 外泌体 成骨分化 骨组织工程 

分 类 号：R622.9[医药卫生—整形外科]