摘要
目的:观察大鼠骨折后骨髓间充质干细胞的组成变化,分析其与骨折愈合的关系。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。选择健康SD大鼠18只,雌雄不限。按随机数字表法分成两组,即一侧股骨骨折组及正常对照组,每组9只。一侧股骨骨折组进行一侧股骨横断性骨折损伤处理,正常对照组不作骨折处理。两组大鼠同等条件下饲养2周后麻醉大鼠,分别在无菌手术条件下取出双侧活体股骨进行骨髓间充质干细胞提取。分离后的细胞分别加入荧光抗体CD44-FITC及CD45-PE,避光孵育30min,加10g/L多聚甲醛重悬固定,冰盒保存,送南华大学流式细胞实验室上机检测,将CD44-FITC阳性同时CD45-PE阴性的细胞定为骨髓间充质干细胞,由此测得的该细胞数值即为骨髓间充质干细胞构成比(指一种细胞数占总细胞数的比值),求其平均值。结果:实验大鼠18只全部进入结果分析。一侧股骨骨折组大鼠处理侧的骨髓间充质干细胞构成比显著高于对侧[29.19±5.06,22.90±7.44(t=2.75,P<0.05)];一侧股骨骨折组大鼠处理侧显著高于正常对照组相应侧(13.05±9.08)(t=3.87,P<0.01);一侧股骨骨折组大鼠处理侧对侧显著高于正常对照组相应侧对侧(14.12±8.98)(t=3.43,P<0.01)。结论:骨折后相应的损伤区域及远隔部位均可出现骨髓间充质干细胞增殖,损伤区域的增殖数量高于远隔部位,骨髓间充质干细胞的构成比增加可能有利于骨折的愈合。
AIM： To study the change of mesenchymal stem cells （MSCs） after bone fracture, to analyze the relationship with fracture healing. METHODS： The experiment was conducted from July 2004 to March 2005 at the Clinical Research Room of the First Affiliated Hospital of Nanhua University. Totally 18 healthy SD rats were selected, of either sex and divided into two groups at random： one-side fracture of femur group and normal control group with 9 rats in each group. The rats in the oneside fracture of femur group received one-side transverse femur fracture. Those in the normal control group did not receive fracture disposal. The rats were anesthetized two weeks after feeding at the same condition of the two groups. The MSCs were isolated from live double：side femurs under aseptic operation. Fluorescent antibody CD44-FITC and CD45-PE were added, incubated for 30 minutes away from light, fixed with 10 g/L formaldehydum polymerisatum, kept in ice container, and delivered to Flow Cell Laboratory, Nanhua University for operating examination. Ceils with positive CD44-FTTC and negative CIMS-PE were identified as MSCs. The numeric value gained from this test was the MSCs constituent ratio （odds of a kind of ceils to total ceils）, and obtained its mean value.
RESULTS： The 18 experimental rats all entered result analysis. The constituent ratio of MSCs at dealing side in ono-side fracture of femur group was significantly higher than that in the other side [29.19±5.06, 22.90±7.44（t=2.75,P〈0.05）]. It was obviously higher in the one-side fracture of femur group than the normal control group （13.05±9.08 ） （t=3.87 ,P〈0.01 ）. it was markedly higher at dealing side of rats in the one-side fracture of femur group than that in the normal control group （14.12±8.98）（t=3.43,P〈0.01）. CONCLUSION： There is proliferation of MSCs at corresponding injured region and further region after bone fracture. The proliferation quantity at injury region is higher than that in the further region. The increase of constituent ratio of MSCs may be helpful for the fracture healing.
出处
《中国临床康复》
CSCD
北大核心
2006年第29期56-58,共3页
Chinese Journal of Clinical Rehabilitation
