CAJ | 학술논문

目的通过体外实验观察兔骨髓基质干细胞(BMSCs)与改良复合磷酸钙骨水泥(BCPC)共培养的生物相容性及诱导成骨活性,探讨其可能的机制.方法配制相同大小规格的BCPC及普通磷酸钙骨水泥(CPC)块,实验分为4组:空白对照组(不加载体)、普通CPC组、BCPC组、加BMP组(加BMP的BCPC).将各组材料与兔BMSCs复合培养,倒置显微镜下观察细胞生长情况,进行细胞计数,碱性磷酸酶(ALP)试剂盒检测细胞合成ALP活性变化,扫描电镜观察细胞在材料表面的生长及长入情况,real-time PCR检测成骨及成软骨相关基因mRNA表达情况并比较表达差别.结果复合培养后倒置显微镜观察各组细胞与载体边缘接触良好,各组细胞增殖差异无统计学意义(P＞0.05);扫描电镜观察细胞在BCPC表面生长增殖良好,逐渐伸出伪足并长入材料孔隙;复合培养7 d后,BCPC组及加BMP组比CPC组和空白对照组表达更高的ALP活性,差异有统计学意义(P＜0.05);复合培养第10天,BCPC组及加BMP组分别与CPC组及空白对照组比较,其成骨及成软骨相关基因mRNA表达差异有统计学意义(P＜0.05).结论配制的BCPC具有良好的生物相容性和一定的成骨诱导能力,其疏松的结构及更大的孔径适宜细胞长入材料内部.
목적 통과체외실험관찰토골수기질간세포(BMSCs)여개량복합린산개골수니(BCPC)공배양적생물상용성급유도성골활성,탐토기가능적궤제.방법 배제상동대소규격적BCPC급보통린산개골수니(CPC)괴,실험분위4조:공백대조조(불가재체)、보통CPC조、BCPC조、가BMP조(가BMP적BCPC).장각조재료여토BMSCs복합배양,도치현미경하관찰세포생장정황,진행세포계수,감성린산매(ALP)시제합검측세포합성ALP활성변화,소묘전경관찰세포재재료표면적생장급장입정황,real-time PCR검측성골급성연골상관기인mRNA표체정황병비교표체차별.결과 복합배양후도치현미경관찰각조세포여재체변연접촉량호,각조세포증식차이무통계학의의(P＞0.05);소묘전경관찰세포재BCPC표면생장증식량호,축점신출위족병장입재료공극;복합배양7 d후,BCPC조급가BMP조비CPC조화공백대조조표체경고적ALP활성,차이유통계학의의(P＜0.05);복합배양제10천,BCPC조급가BMP조분별여CPC조급공백대조조비교,기성골급성연골상관기인mRNA표체차이유통계학의의(P＜0.05).결론 배제적BCPC구유량호적생물상용성화일정적성골유도능력,기소송적결구급경대적공경괄의세포장입재료내부.
Objective To investigate in vitro the biocompatibility of modified calcium phosphate bone cement(BCPC) and rabbit bone mesenchymal stem cells (BMSCs) as well as the effect of the composite on the growth and osteogenic differentiation of rabbit BMSCs. Methods The blocks of BCPC and ordinary calcium phosphate cement (CPC) were prepared in the same size. The experiment was conducted in 4 groups: ordinary CPC group, BCPC group, BCPC plus bone morphogenetic protein (BMP) group and blank control group.Rabbit BMSCs were co-cultured with the 4 vectors respectively. Cell growth was observed by inverted phase microscopy. Changes in the alkaline phosphatase(ALP) activity were detected by an ALP test kit. The BMSCs growth on the surface of the vectors was observed by scanning electron microscopy. Real-time PCR was used to measure the expressions of relative gene mRNA for osteogenesis and chondroblasts. Results There was no statistical difference in cell proliferation among the 4 groups( P ＞ 0.05 ). The BMSCs contacted well with the vector edges and grew well on the surface of the vectors. Scanning electron microscopy showed the pseudopodia extended from the cells into the pore of BCPC. Higher activity of ALP was detected in the BMP and BCPC groups than in the other 2 groups 7 days after co-culturing( P ＜ 0.05 ). On the 10th day after co-culturing, the expressions of relative gene mRNA for osteogenesis and chondroblasts were significantly higher in the BCPC and BMP groups than in the control group( P ＜ 0.05 ). Conclusions The BCPC has a good biocompatibility with BMSCs and can induce osteoblasts to a certain degree. The porous structure and large apertures of the vector enable cells to grow into its deep layers.