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

[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。

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

[Objective]To evaluate cell adhesion and cellular compatibility of six new composite scaffolds made from PDLLA,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/β-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,PDLLA/β-TCP and PDLLA/PLA-PEG-PLA than that of PDLLA cell-polymer.Among these polymers,PDLLA/PLA-PEG-PLA 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.