CAJ | 학술논문

目的研究异种骨材料对骨髓间充质干细胞(BMSC)的增殖、迁移、细胞周期及定向分化为成骨细胞的能力,并对其生物安全性进行评价.方法取体外培养第5代骨髓间充质干细胞直接接触培养,观察经环氧化物固定制备的异种骨浸提液对骨髓间充质干细胞的增殖率、迁移率、细胞周期、碱性磷酸酶(ALP)及钙结节形成的影响,以H-DMEM培养液组作为阴性对照.结果扫描电镜观察BMSC于异种骨上生长状态饱满,细胞呈梭形或多边形；骨髓间充质干细胞在浸提液中培养,第1、3、5、7d组与对照组相比,结果差异无统计学意义,无细胞毒性；膜滤器(Transwell小室)检测细胞迁移,浸提液组为(46.00±5.00)个/视野,对照组为(37.00±3.80)个/视野,P＜0.05.浸提液组组织在G2/M期的细胞为(27.77±1.06)％,对照组为(26.60±1.05)％,P＜0.01.浸提液组的ALP活性高于对照组(P＜0.05),茜素红染色显示浸提液组于14 d时钙沉积明显增多.结论所制备异种骨具有良好的细胞相容性,并对骨髓间充质干细胞的迁移和成骨分化有一定的促进作用.
목적 연구이충골재료대골수간충질간세포(BMSC)적증식、천이、세포주기급정향분화위성골세포적능력,병대기생물안전성진행평개.방법 취체외배양제5대골수간충질간세포직접접촉배양,관찰경배양화물고정제비적이충골침제액대골수간충질간세포적증식솔、천이솔、세포주기、감성린산매(ALP)급개결절형성적영향,이H-DMEM배양액조작위음성대조.결과 소묘전경관찰BMSC우이충골상생장상태포만,세포정사형혹다변형；골수간충질간세포재침제액중배양,제1、3、5、7d조여대조조상비,결과차이무통계학의의,무세포독성；막려기(Transwell소실)검측세포천이,침제액조위(46.00±5.00)개/시야,대조조위(37.00±3.80)개/시야,P＜0.05.침제액조조직재G2/M기적세포위(27.77±1.06)％,대조조위(26.60±1.05)％,P＜0.01.침제액조적ALP활성고우대조조(P＜0.05),천소홍염색현시침제액조우14 d시개침적명현증다.결론 소제비이충골구유량호적세포상용성,병대골수간충질간세포적천이화성골분화유일정적촉진작용.
cell cycle and osteoblastic differentiation of bone marrow derived stromal cells (BMSCs),and evaluate the biosafety of heterogeneous bone.Methods The heterogeneous bones were fixed with epoxides and the extracts were prepared according to ISO 10993-12:2007.Mouse passage 5 BMSCs were cultured with the heterogeneous bone extracts for 24 hours to observe cell proliferation,migration,cell cycle,ALP activity and mineralization node formation.The morphology of BMSCs was observed by scanning electron microscopy (SEM).Cells cultured with H-DMEM without addition of the extracts served as control.Results Morphological observation under SEM showed healthy growth of BMSCs on heterogeneous bone extracts with spindle-shape or polygonal shape.There was no obvious difference in cell proliferation after 1,3,5 and 7 days co-culture with the extracts when compared to the control,indicating the absence of cytotoxicity.The transwell cell migration test showed (46.00±5.00) cells/field in the extract group and (37.00 ± 3.80) cells/field in the control group (P ＜ 0.05).The extract group induced cell cycle arrest at G2/M phase in about (27.77 ± 1.06)％ of the cells,while the control group (26.60 ± 1.05) ％ (P ＜ 0.01).ALP activity and mineralization nodes detected by Alizarin bordeaux staining were significantly higher in the extract group than in the control group after 14 days' co-culture (P ＜ 0.05).Conclusion The heterogeneous bone was biocompatible.It can potentially promote BMSC migration and osteoblastic differentiation.