鼻黏膜来源外胚间充质干细胞与磷酸钙骨水泥材料的体外相容性  

Compatibility of nasal mucosa-derived ectomesenchymal stem cells and calcium phosphate cement in vitro

作　　者：史文涛 戴瑶 陈平波 毕士奇 吕德明 陆浩 张志坚 Shi Wen-tao;Dai Yao;Chen Ping-bo;Bi Shi-qi;Lü De-ming;Lu Hao;Zhang Zhi-jian(Department of Orthopedics,Gaochun People’s Hospital of Nanjing,Nanjing 211300,Jiangsu Province,China;School of Medicine,Jiangsu University,Zhenjiang 212000,Jiangsu Province,China)

机构地区：[1]南京市高淳人民医院骨科,江苏省南京市211300 [2]江苏大学医学院,江苏省镇江市212000

出　　处：《中国组织工程研究》2018年第33期5350-5355,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金地区科学基金资助项目(81571830);中文名称:TG2/SHH基因修饰EMSCs-Fibrin支架对NSCs命运调控机制及修复脊髓损伤研究~~

摘　　要：背景:利用磷酸钙骨水泥载间充质干细胞治疗骨缺损已取得令人满意的成果,但是鼻黏膜来源的外胚层间充质干细胞与磷酸钙骨水泥的体外相容性尚不清楚。目的:探讨磷酸钙骨水泥对大鼠鼻黏膜来源外胚间充质干细胞向成骨细胞分化的影响,进一步探讨鼻黏膜来源外胚间充质干细胞与磷酸钙骨水泥的生物相容性。方法:体外培养扩增鼻黏膜来源外胚间充质干细胞,并将其种植于磷酸钙骨水泥表面和培养板(对照组),均进行成骨诱导14 d。碱性磷酸酶活性法检测鼻黏膜来源外胚间充质干细胞的碱性磷酸酶活性,茜素红染色法测定钙结节面积大小,免疫荧光和免疫印迹法检测成骨诱导相关蛋白表达水平;鬼笔环肽染色法动态观察磷酸钙骨水泥表面的细胞形态;MTT法检测磷酸钙骨水泥对细胞增殖的影响。结果与结论:(1)成骨诱导14 d后,磷酸钙骨水泥表面的鼻黏膜来源外胚间充质干细胞碱性磷酸酶活性较高,形成的钙结节较大,骨相关蛋白表达水平较高;(2)磷酸钙骨水泥对鼻黏膜来源外胚间充质干细胞的增殖有一定的促进作用;(3)结果表明,鼻黏膜来源外胚间充质干细胞与磷酸钙骨水泥支架材料具有良好的生物相容性,磷酸钙骨水泥对鼻黏膜来源外胚间充质干细胞向成骨细胞分化表现出一定的促进作用。BACKGROUND:The use of calcium phosphate cement(CPC)carrying mesenchymal stem cells for the treatment of bone defects has achieved satisfactory results.The in vitro compatibility of ectomesenchymal stem cells(EMSCs)derived from the nasal mucosa with CPC is unknown.OBJECTIVE:To investigate the effect of calcium phosphate cement on the differentiation of rat nasal mucosa-derived EMSCs into osteoblasts and to further explore the biocompatibility of EMSCs and CPC.METHODS:EMSCs were cultured,expanded in vitro and implanted onto the CPC surface or onto culture plates.Then,implanted EMSCs were induced to differentiate into osteoblasts for 14 days.Alkaline phosphatase activity assay was used to detect the alkaline phosphatase activity of EMSCs.The area of calcium nodules was determined by alizarin red staining.The expression level of osteogenesis-related proteins was detected by western blot assay and immunofluorescence method.Morphology of EMSCs on the CPC surface was dynamically observed using phalloidin staining.MTT assay was used to detect the proliferation of EMSCs on the CPC surface.RESULTS AND CONCLUSION:After 14 days of osteogenic induction,there was higher alkaline phosphatase activity,larger calcium nodules and higher levels of osteogenesis-related proteins in the EMSCs implanted on the surface of CPC than those implanted on the culture plates.The CPC had a certain role in promoting the proliferation of EMSCs.To conclude,EMSCs and CPC scaffolds have good biocompatibility,and moreover,CPC can certainly promote the differentiation of EMSCs into osteoblasts.

关 键 词：鼻黏膜 外胚间充质干细胞 磷酸钙骨水泥 成骨分化 生物相容性 干细胞 国家自然科学基金 鼻黏膜 间质干细胞 磷酸钙类 细胞分化 组织工程 

分 类 号：R394.2[医药卫生—医学遗传学]