恒河猴间充质干细胞诱导分化成骨细胞复合6种支架材料的相容性研究  

Experimental study on cellular compatibility of six kinds of new scaffolds with osteoblasts differentiated from MSCs of rhesus monkey

作　　者：闵少雄[1] 李森[2] 刘成龙[1] 曹延林[1] 张力[1] 靳安民[1] 

机构地区：[1]南方医科大学附属珠江医院骨科中心,广州510282 [2]泸州医学院附属医院脊柱骨科,泸州646000

出　　处：《中国矫形外科杂志》2012年第15期1420-1423,共4页Orthopedic Journal of China

基　　金：广东省科技计划项目(编号:2010B080701019)

摘　　要：[目的]评价外消旋聚乳酸(poly-DL-lactide,PDLLA)、生物活性玻璃(bioactive glass,BG)、β-三磷酸钙(beta-tricalcium phosphate,β-TCP)与聚乳酸-聚乙二醇-聚乳酸三嵌段共聚物[poly(lactic acid)-poly-ethylene glycol-poly(lactic acid)triblock copolymers,PLA-PEG-PLA]组合成的6种新型支架的细胞粘附性和细胞相容性,筛选出较好的细胞支架。[方法]选用恒河猴间充质干细胞(mesenchymal stem cells,MSCs)定向分化为成骨细胞,采用体外细胞培养的方法,将细胞接种于PDLLA、PDLLA/BG、PDLLA/β-TCP、PDLLA/PLA-PEG-PLA、PDLLA/PLA-PEG-PLA/BG及PDLLA/PLA-PEG-PLA/β-TCP 6种可吸收性生物支架材料表面上,倒置显微镜、扫描电镜(scanning electron microscope,SEM)、碱性磷酸酶(alkaline phosphatase,ALP)染色及甲基偶氮唑盐微量酶反应比色(methyl thiazolyl tetrazolium,MTT)法观察恒河猴成骨细胞的粘附、增殖。[结果]倒置显微镜、SEM、ALP染色显示成骨细胞在PDLLA、PDLLA/PLA-PEG-PLA、PDLLA/β-TCP及PDLLA/BG 4种材料上能良好粘附、增殖,并向内部生长,而在PDLLA/PLA-PEG-PLA/BG、PDLLA/PLA-PEG-PLA/β-TCP两组材料上粘附差、细胞逐渐死亡;MTT法结果显示PDLLA/PLA-PEG-PLA、PDLLA/β-TCP及PDLLA/BG细胞活性优于单纯PDLLA支架,其中PDLLA/PLA-PEG-PLA最佳。[结论]PDLLA、PDLLA/PLA-PEG-PLA、PDLLA/BG及PDLLA/β-TCP4组材料能与成骨细胞良好粘附,具有良好的细胞相容性。[ Objective ] To evaluate cell adhesion and cellular compatibility of six new composite scaffolds made from PDL- LA, BG, β -TCP and PLA -PEG -PLA, find good scaffold for bone tissue engineering. [ Methods ] The osteoblasts differentiated from MSCs of rhesus monkey were used to estimate osteocompatibility of six kinds of new scaffolds in vitro. The cells were inoculated onto of PDLLA, PDLLA/BG, PDLLA/β- TCP, PDLLA/PLA - PEG - PLA, PDLLA/PLA - PEG - PLA/BG and PDLLA/PLA -PEG- PLA/β-TCP scaffolds. Inverted microscope, SEM, ALP staining and MTT assay were used to evaluate the adhesiveness and proliferation of rhesus monkey osteoblasts. [ Results ] Inverted microscope, SEM, ALP staining demonstrated that osteoblasts on PDLLA, PDLLA/BG, PDLLA/13 - TCP and PDLLA/PLA - PEG - PLA 4 kinds of polymers had better adhesion, proliferation, and internal growth. Comparatively, osteoblasts on PDLLA/PLA - PEG - PLA/BG and PDLLA/PLA - PEG - PLA/β - TCP cell - polymers had poor adhesion and cell died gradually. The results of MTT showed better cell viability of PDLLA/BG, PDLLAβ- TCP and PDLLA/PLA - PEG - PLA than that of PDLLA cell - polymer. Among these polymers, PDLLA/PLA- PEG- PEA showed the best results of osteoblasts adhesion, proliferation and viability. [ Conclusion] PDLLA, PDLLA/BG, PDLLA/β -TCP and PDLLA/PLA- PEG- PLA scaffolds showed good cellular compatibility and cell adhesion.

关 键 词：细胞相容性 聚乳酸 外消旋聚乳酸 聚乙二醇 间充质干细胞 成骨细胞 

分 类 号：R318.08[医药卫生—生物医学工程]