骨损伤刺激对骨髓间充质干细胞增殖的影响

Influence of bone injury on the proliferation of mesenchymal stem cells

目的:观察骨损伤性刺激对骨髓间充质干细胞的影响,为体内诱导骨髓间充质干细胞增殖提供方法。方法:实验于2004-07/2005-03在南华大学附属第一医院临床研究所完成。选取健康SD大鼠18只,每组9只,随机分成两组,一组为观察组,进行一侧闭合性股骨横断性骨折损伤模型处理,另一组为对照组不作骨折损伤处理,两组大鼠同等条件下饲养2周,然后取出双侧股骨进行骨髓间充质干细胞提取,流式细胞仪检测含CD44-FITC阳性而CD45-RPE阴性的细胞的构成比(骨髓间充质干细胞具有表现为CD44阳性而及CD45阴性的特点,因此把通过流式细胞仪检测到的CD44-FITC阳性同时CD45-RPE阴性的细胞认定为骨髓间充质干细胞,由此测得的该细胞数占总检测细胞数的比为骨髓间充质干细胞构成比),计算出两组骨髓间充质干细胞构成比均数。结果:实验大鼠18只均进入结果分析。①观察组处理侧骨髓间充质干细胞构成比均数高于对照组处理侧(29.19±5.06,13.05±9.08,P<0.01)。②观察组未处理侧股骨骨髓间充质干细胞构成比均数高于对照组未处理侧(22.90±5.44,14.12±8.98,P<0.01)。③观察组处理侧股骨骨髓间充质干细胞构成比均数低于未处理侧(29.19±5.06,22.90±5.44,P<0.05)。结论:骨损伤后骨髓间充质干细胞出现增殖,为骨折的愈合提供物质基础,提示骨损伤性刺激可作为活体内诱导骨髓间充质干细胞增殖的一种方法。

AIM： To observe the effect of bone injury on proliferation of mosenchymal stem cells （MSCs） so as to provide the method for inducing the proliferation of MSCs. METHODS： This experiment was Carried out at the Clinical Institute of First Hospital Affiliated to Nanhua University from July 2004 to March 2005. Totally 18 healthy SD rats were chosen and randomly divided into 2 groups with 9 in each group： observation group, in which, rats underwent one side closed femoral traverse fracture injury and control group, in which, rats were not treated, Rats in the two groups were raised for 2 weeks at the same condition. Then, bilateral femoral bone was taken out to extract MSCs, The constituent ratio of the cells containing CD44-FITC positive and CD45-RPE negative was detected with flow cytometer （MSCs were characterized by expressing CD44 positive and CIM5 negative, therefore,, cells containing CD44-FITC positive and CD45-RPE negative detected by flow cytometer were concluded as MSCs, thus, the ratio of the number between detected cells and total detected cells was the constituent ratio of MSCs）, The mean value of constituent ratio of the BMCs of two groups was calculated, RESULTS： All the 18 rats entered the stage of result analysis, （1） Constituent ratio of MSCs of treated side was higher in observation group than in control group （29.19 ±5,06,13.05 ±9,08,P 〈 0.01）, （2） Constituent ratio of MSCs of femoral bone was higher in treated side of observation group than in untreated side of control group （22.90±5.44, 14.12±8.98 ,P 〈 0.01）, （3） The mean value of constituent ratio of MSCs of femoral bone was lower in treated side than in untreated side of observation group （29.19±5.06,22.90±5.44,P 〈 0.05） CONCLUSION： Proliferation of MSCs appears after bone injury, which provides material basis for the healing of fracture, suggesting that bone traumatic stimulus can be taken as a method in inducing the proliferation of MSCs in vivo.