欢迎访问《解剖学报》官方网站！今天是
English
骨髓间充质干细胞与阿魏酸处理的PHBV膜的体外相容性
Compatibility study of bone mesenchymal stem cells and ferulic acid treatment PHBV film in vitro
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。
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 BMSCsin 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.
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
Bone mesenchymal stem cells / 3-(4-hydroxy-3-methoxyphenyl)-2-propenoic acid / Ferulic acid / Bone tissue engineering / Scanning electron microscopy / Rat
［1］Sendil D, Gürsel I, Wise DL, et al. Antibiotic release from biodegradable PHBV microparticles［J］. J Contr Rel, 1999,59(2):207-217.
［2］Chen J, Davis SS. The release of diazepam from poly(hydroxybutyrate-hydroxyvalerate) microspheres［J］. J Microencapsulation, 2002,19（2）:191-201.
［3］Chen GQ,Wu Q.The application of polyhydroxyalkanoates as tissue engineering materials［J］.Biomaterials，2005，26(33)：6565-6578.
［4］Li H，Chang J.Fabrication and characterization of bioactive wollastonite/PHBV composites scaffolds［J］. Biomaterials，2004，25（24）：5473-5480.
［5］Choi J S, Lee S W, Jeong L, et al. Effect of organosoluble salts on the nanofibrous structure of electrospun poly(3-hydroxybutyrate-co-3-hydroxyvalerate) ［J］. Int J Biol Macromol, 2004, 34(4):249-256.
［6］Tezcaner A，Bugra K,Hasici V.Retinal pigment epithelium cell culture on surface modified poly thin films［J］.Biomaterials, 2003, 24(25):4573-4583.
［7］Volova TG, Shishatskaya EI, Sevastianov V, et al.Results of biomedical investigations of PHB and PHB/PHV fibers［J］. Biochem Eng J，2003，16（2）：125-133.
［8］Zuo L,Bai ShL， Pan F, et al.The compatibility study of osteoblast and the silicate-phosphate coating on the surface of the magnesiumin vitro［J］.Acta Anatomica Sinica, 2011，42(1):65-69.（in Chinese）
左林，柏树令，潘锋，等.成骨细胞与镁合金表面硅酸盐-磷酸盐复合涂层的体外相容性研究［J］.解剖学报，2011，42(1):65-69.
［9］Zhang L,Bai SL, Pan F,et al. Compatibility of human umbilical vein endothelial cells and surface-modified PHBHHx film in vitro［J］. Acta Anatomica Sinica, 2009，6(9):10-13. (in Chinese)
张磊，柏树令，潘锋，等.人脐静脉内皮细胞与表面改性的PHBHHx膜的体外相容性研究［J］.解剖学报，2009，6(9):10-13.
［10］Peister A, Mellad JA, Larson BL, et al. Adult stem cells from bone marrow (MSCs) isolated from different strains of inbred mice vary in surface epitopes, rates of proliferation, and differentiation potential［J］. Blood, 2004,103(5):1662-1668.
［11］Yang L,Zhang RH,Xie HJ,Cai Y,et al. Establishment of a stable isolation culture system and identification of rat bone marrow mesenchymal stem cells［J］. Journal of Clinical Rehabilitative Tissue Engineering Research, 2009;13(6):1064-1068. (in Chinese)
杨丽,张荣华,谢厚杰,等.建立大鼠骨髓间充质干细胞稳定分离培养体系与鉴定［J］.中国组织工程研究与临床康复,2009, 13(6):1064-1068.
［12］Yang YJ，Li ZhZh，Zhang XQ. Present research of using PHAs synthesized by microbes［J］. Chemical Research and Application, 2006,18（9）. (in Chinese)
羊依金，李志章，张雪乔.微生物降解塑料的研究进展.［J］化学研究与应用，2006,18（9）
［13］Tamilvanan, S, Venkateshan N, Ludwig A. The potential of lipid-and polymer-based drug delivery carriers for eradicating biofilm consortia on device-related nosocomial infections［J］.J Control Release，2008，128（1）：2-22.
［14］Wang J，Zhang HY,Hu Y,et al.In vitro studies of bone marrow mesenchymal stem cells on HA/TCP bone graft scaffold ［J］.Acta Anatomica Sinica,2008,39(4):539-542. (in Chinese)
王健，张海燕，胡勇,等.骨髓间充质干细胞复合羟基磷灰石/磷酸三钙植骨材料的体外研究［J］.解剖学报,2008,39(4):539-542.
国家自然科学基金重点资助项目
/
| 〈 |
|
〉 |
