过表达BMP-12的兔BM-MSCs在3D打印PLGA支架上的增殖与分化特性研究  

Proliferation and differentiation of BMP-12 overexpressed rabbit BM-MSCs on 3D printed PLGA scaffolds

作　　者：崔磊 陈鹏[2] 张文涛[2] 谢小肖 李伟[2] 江长青 CUI Lei;CHEN Peng;ZHANG Wentao(Clinical College of Peking University Shenzhen Hospital,Anhui Medical University,Shenzhen,Guangdong,518036,China;不详)

机构地区：[1]安徽医科大学北京大学深圳医院临床学院,广东深圳518036 [2]北京大学深圳医院运动医学科

出　　处：《实用手外科杂志》2021年第3期368-374,共7页Journal of Practical Hand Surgery

基　　金：深圳市科技计划项目(项目编号:20180214121124778);深圳市医疗卫生三名工程(项目编号:SZSM201612078)。

摘　　要：目的探讨过表达骨形态发生蛋白-12(Bone Morphogenic Protein 12,BMP-12)的兔骨髓间充质干细胞(Bone Marrow-derived Mesenchymal Stem Cells,BM-MSCs)在3D打印聚乳酸-羟基乙酸(Polylactic-co-glycolic Acid,PLGA)支架上的增殖与分化特性。方法采用3D打印技术制备PLGA支架,并检测其降解性能,分离、培养兔BM-MSCs,使用BMP-12重组腺病毒表达载体转染兔BMMSCs,并将其种植在PLGA支架上,检测支架的细胞毒性及种植的细胞在支架上的增殖活性,通过实时荧光定量PCR和Westernblot技术检测种子细胞Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、BMP-12、SCX、Tnmd、Tnc的基因和蛋白表达水平。结果PLGA支架具有良好的生物相容性和生物可降解性。在种植了过表达BMP-12的BM-MSCs的3 D打印PLGA支架上,BM-MSCs的Ⅰ型胶原蛋白、Ⅲ型胶原蛋白、BMP-12、SCX、Tnmd、Tnc的mRNA和蛋白表达水平显著升高。结论本研究结果提示,在携带过表达BMP-12的BM-MSCs的3 D打印PLGA支架上,BMP-12的过表达促进了BM-MSCs的腱向分化。携带过表达BMP-12的BM-MSCs的PLGA支架可以为肌腱损伤修复提供一种新的治疗方法。Objective To investigate the proliferation and differentiation of rabbit bone marrow mesenchymal stem cells(BM-MSCs) overexpressing bone morphogenetic protein-12(BMP-12) on 3D printed PLGA scaffolds. Methods PLGA scaffolds was prepared by 3D printing technology. Bone marrow mesenchymal stem cells were isolated and cultured. BMP-12 recombinant adenovirus expression vector was used to transfect rabbit BM-MSCs. The cytotoxicity of the scaffolds and the proliferation of the cells on the PLGA scaffolds were detected. The protein expression levels of type Ⅰ collagen, type Ⅲ collagen,BMP-12, SCX, Tnmd and Tnc were detected by real-time fluorescent quantitative PCR and Western blot. Results PLGA scaffolds had good biocompatibility and biodegradability. On the 3D printed PLGA scaffolds with BMP-12 overexpressed BM-MSCs, the mRNA and protein expression levels of type I collagen, type Ⅲ collagen, BMP-12, Scx,Tnmd and Tnc in BM-MSCs were significantly increased.Conclusion These results suggest that overexpression of BMP-12 promotes tendon differentiation of BMMSCs on 3 D printed PLGA scaffolds. 3D printed PLGA scaffolds carrying BMP-12 overexpressed BM-MSCs provides a new treatment for tendon injury.

关 键 词：骨髓间充质干细胞 PLGA支架 BMP-12 肌腱修复 3 D打印 

分 类 号：R686.1[医药卫生—骨科学] R318.08[医药卫生—外科学]