利福平对兔脂肪干细胞增殖、成骨及黏附支架的影响  

Effects of Rifampicin on the Proliferation,Osteogenic Differentiation and Adhesion to Scaffold of Rabbit Adipose-derived Stem Cells

作　　者：梁求真 吐汉.艾买提 宋兴华[1] 蒋岳华[1] 马艳[3] 毕晓娟[3] 白晶晶[1] 罗兰[4] 

机构地区：[1]新疆医科大学第一附属医院骨科中心,乌鲁木齐830054 [2]新疆医科大学第一附属医院呼吸科,乌鲁木齐830054 [3]新疆医科大学第一附属医院临床医学研究院干细胞研究中心,乌鲁木齐830054 [4]新疆医科大学第一附属医院药学部,乌鲁木齐830054

出　　处：《医学研究杂志》2014年第2期27-31,共5页Journal of Medical Research

基　　金：国家自然科学基金资助项目(81360283)

摘　　要：目的探讨利福平（rifampicin，RFP）对兔脂肪干细胞（rabbitadipose-derivedstemcells，rADSCs）增殖、成骨及黏附支架的影响，为后期RFP缓释系统与rADSCs的复合奠定实验基础。方法分离培养rADSCs，检测细胞周期及表面抗原CD44，成骨、成脂诱导后分别行茜素红及油红O染色。用利福平生长液干预rADSCs，按浓度分为0、10、20及30μg／ml4组，分别绘制细胞生长曲线、检测细胞凋亡率及碱性磷酸酶（alkalinephosphatase，ALP）含量。制备同种异体脱钙骨支架，并复合各组rADSCs，计算细胞黏附率，扫描电子显微镜观察复合情况。结果rADSCs呈长梭形生长，G0／G1期占78．3％±4．2％，表面抗原CD44阳性表达。成骨诱导后茜素红染色及成脂诱导后油红O染色均为阳性。生长曲线示30μg／ml组第4-11天增殖能力较其他3组弱（P〈0．01），细胞凋亡示30μg／ml组凋亡率高于其他3组（P〈0．01），ALP检测示成骨诱导后第14天30μg／ml组ALP含量较其他3组少（P〈0．01），细胞黏附率示30μg／ml组较其他3组低（P〈0．01）。扫描电镜观察30μg／ml组黏附的细胞数量少，形态松散。结论不影响rADSCs增殖、成骨及黏附支架的RFP临界浓度约为20μg／ml。Objective To evaluate the effects of rifampicin （RFP） on the proliferation, osteogenic differentiation and adhesion to scaffold of rabbit adipose - derived stem cells（ rADSCs）. Methods The adipose tissue was obtained from inguinal fat pads of the rabbit,and digested with type I collagenase. The characteristics of rADSCs were certified by the results of cell cycle, anti - CD44 antibody and multipotential differentiation including osteogenic induction and adipogenic induction. The rADSCs were randomly divided into four group according to drug concentration of rifampicin, including group 0μg/ml, group 10μg/ml, group 20μg/ml and group 30μg/ml. The cell growth curves, cell apoptosis and alkaline phosphatase（ ALP） activity were used for determining the effects of rifampicin on rADSCs. De- calcified allograft bone scaffold were prepared, and the adhesion rate was calculated after inoculation. The appearance of the complex was also observed by scanning electron microscope（SEM）. Results The rADSCs grew spindle - shaped in vitro, G0/G1, cycle was around 78.3% ＋ 4.2%. Expression of CD44 was positive. Alizarin red staining demonstrated positive formation of mineralized nods. The results of Oil red O staining was also positive. The results of cell growth curves showed the proliferation in group 30μg/ml were lower than that in other groups from day 4 to 11. The cell apoptosis rate was higher in group 30μg/ml than that in other groups（P 〈 0.01 ）. The expression of ALP was fewer on day 14 in group 30μg/ml than that in other groups（P 〈 O. 01 ）. The adhesion rate was lower in group 30μg/ml than that in other groups （ P 〈 0.01 ）. The appearance of SEM showed that in group 30 μg/ml the cells were fewer and looser. Conclusion The critical concentration of RFP without effects on the proliferation, osteogenic differentiation and adhesion to scaffold of rADSCs was about 20μg/ml.

关 键 词：骨结核 脂肪干细胞 利福平 

分 类 号：R68[医药卫生—骨科学]