解剖学报

解剖学报 ›› 2014, Vol. 45 ›› Issue (1): 47-52.doi: 10.3969/j.issn.0529-1356.2014.01.009

• 细胞和分子生物学 • 上一篇    下一篇

骨髓间充质干细胞与阿魏酸处理的PHBV膜的体外相容性

宋章硕1 柏树令1* 潘峰2 田晓红1 佟浩1   

  1. 1.中国医科大学组织工程研究所; 2.中国医科大学人体解剖学教研室， 沈阳 110001
  • 收稿日期:2013-01-04 修回日期:2013-09-11 出版日期:2014-02-06 发布日期:2014-02-06
  • 通讯作者: 柏树令 E-mail:baishuling@hotmail.com
  • 基金资助:

    国家自然科学基金重点资助项目

Compatibility study of bone mesenchymal stem cells and ferulic acid treatment PHBV film in vitro

SONG Zhang-shuo1 BAI Shu-ling 1* PAN Feng2 TIAN Xiao-hong1 TONG Hao1   

  1. 1.Department of Tissue Engineering，College of Basic Medical Sciences，China Medical University，Shenyang 110001，China;2. Department of Human Anatomy, College of Basic Medical Sciences，China Medical University，Shenyang 110001，China
  • Received:2013-01-04 Revised:2013-09-11 Online:2014-02-06 Published:2014-02-06
  • Contact: BAI Shu-ling E-mail:baishuling@hotmail.com

摘要：

目的 观察骨髓间充质干细胞（BMSCs）在含有不同浓度3-(4-羟基-3-甲氧基苯基)-2-丙烯酸（阿魏酸）处理的3-羟基丁酸-co-3-羟基戊酸共聚物（PHBV）膜上的生长情况，探讨PHBV膜与BMSCs的生物相容性，为骨组织工程支架材料的选择提供依据。方法 用全骨髓培养法分离培养大鼠BMSCs，流式细胞仪检测细胞表面抗原，进行鉴定。将第3代大鼠 BMSCs接种于材料表面，分别培养8h、12h、24h后，用扫描电镜观察，BMSCs在不同浓度阿魏酸处理的材料表面上的黏附的形态变化，制备浸提液，用浸提液进行细胞培养。用MTT法和细胞周期法检测材料对细胞的影响。结果 成功培养出BMSCs，流式细胞仪检测CD44、CD90表达结果呈阳性，CD34、CD11b表达结果呈阴性。扫描电镜下可见，PHBV材料表面粗糙，可见结晶体。将BMSCs与材料复合培养后，可见不含阿魏酸和含5%阿魏酸组材料表面的细胞形态规则、分布均匀、活力旺盛。MTT分析结果和细胞周期检测显示，经5%阿魏酸处理的PHBV膜对大鼠BMSCs的体外生长和增殖的促进作用最佳。 结论 BMSCs与阿魏酸处理的PHBV膜具有良好的生物相容性，体外培养环境下有利于细胞的生长、黏附和增殖。

关键词: 骨髓间充质干细胞, PHBV, 3-（4-羟基-3-甲氧基苯基)-2-丙烯酸, 骨组织工程, 扫描电镜, 大鼠

Abstract:

Objective To evaluate the growing condition of bone mesenchymal stem cells (BMSCs) which were cultivated in the film of different concentration in the ferulic acid treatment of abamectin poly-3-hydroxy butyrate-co-3-hydroxy valerate(PHBV), and to investigate biocompatibility between the membrane of poly-3-hydroxy butyrate-co-3-hydroxy valerate and bone mesenchymal stem cells to provide a basis for the choice of scaffold material of bone tissue engineering. Methods The BMSCs were harvested by a whole bone marrow culture method,and identified by flow cytometric to analysis detection cell surface antigen. The BMSCs of the third passage were inoculated on the material surface and cultured for 8 hours,12 hours and 24 hours. Changes in form of BMSCs on different concentrations in the ferulic acid treatment surfaces were observed by scanning electron microscopy(SEM).The material leaching liquor to culture BMSCs was prepared and material was detected with the influence of cells by MTT and flow cytometer cell cycle assays method. Results The BMSCs were successfully cultured, these cells were uniformly negative for CD44，CD90 and positive for CD34，CD11b.SCM showed that PHBV material surface coarse, visible crystals.The cells on the group of a and b material surface growed well,and proliferated significantly.The MTT analysis and flow cytometer cell cycle assays showed that the PHBV film of 5% ferulic acid treatment promoted the growth and proliferation of BMSCsin vitro. Conclusion The PHBV film with 5% ferulic acid treatment had good biocompatibility of BMSCs. Environment is conducive to cell growth, adhesion and proliferation in vitro.

Key words: Bone mesenchymal stem cells, 3-(4-hydroxy-3-methoxyphenyl)-2-propenoic acid, Ferulic acid, Bone tissue engineering, Scanning electron microscopy, Rat