CAJ | 학술논문

目的：观察域型糖尿病大鼠颌骨骨髓基质干细胞(BMSCs)的成骨分化特性。方法：无菌条件下从10周龄GK大鼠(实验组)及同龄正常Wistar大鼠(对照组)下颌骨获取BMSCs，采用全骨髓贴壁培养法对细胞进行纯化及培养，流式细胞仪检测干细胞表型.取第3代细胞成骨诱导培养，分别对两组细胞行茜素红染色，碱性磷酸酶染色及活性测定，培养基钙、磷含量测定，以及real-time PCR测定Runx2基因表达变化。结果：成功培养出域型糖尿病大鼠颌骨来源的BMSCs，与正常组比较，实验组BMSCs茜素红染色变浅、钙结节形成减少，碱性磷酸酶(ALP)活性下降，细胞钙、磷分泌显著降低，差异有统计学意(P＜0.05)；Runx2 mRNA表达也明显降低(P＜0.05)。结论：域型糖尿病大鼠颌骨来源的BMSCs的成骨分化能力受到损害，并且即使在正常糖浓度下培养其成骨能力仍无法恢复。
목적：관찰역형당뇨병대서합골골수기질간세포(BMSCs)적성골분화특성。방법：무균조건하종10주령GK대서(실험조)급동령정상Wistar대서(대조조)하합골획취BMSCs，채용전골수첩벽배양법대세포진행순화급배양，류식세포의검측간세포표형.취제3대세포성골유도배양，분별대량조세포행천소홍염색，감성린산매염색급활성측정，배양기개、린함량측정，이급real-time PCR측정Runx2기인표체변화。결과：성공배양출역형당뇨병대서합골래원적BMSCs，여정상조비교，실험조BMSCs천소홍염색변천、개결절형성감소，감성린산매(ALP)활성하강，세포개、린분비현저강저，차이유통계학의(P＜0.05)；Runx2 mRNA표체야명현강저(P＜0.05)。결론：역형당뇨병대서합골래원적BMSCs적성골분화능력수도손해，병차즉사재정상당농도하배양기성골능력잉무법회복。
0bjective:To observe osteogenic differentiation ability of bone marrow stromal cells (BMSC) obtained from type 2 diabetic rats mandibles.Methods:The invitrotechnique of cell culture was employed to harvest, purify and cultivate BMSCs from the mandibles of type 2 diabetic(GK, test group) and normal rats (Wistar,control group) . Flow cytometry analyses of cell phenotype and osteogenic/adipogenic differentiation were performed. The passage 3 cells were incubated with osteogenic DMEM. The cells were then stained with 2% Alizarin red S. The Alizarin red S-stained nodules were observed microscopically and quantified. ALP staining and ALP activity assay were performed. The calcium content and phosphonium content were evaluated. The expression ofRUNX2mRNA was analysed by real-time PCR. The data were subjected tot-testusing SPSS software.Results:BMSCs were obtained from the mandibles of type 2 diabetic rats successfully. Compared with BMSCs derived from normal rats, the number of red calcium nodules in the test group were obviously decreased, ALP activity was inhibited and the contents of calcium and phosphonium in medium were also decreased. The difference was statistically significant (P<0.05). Furthermore,RUNX2mRNA expression was decreased (P<0.05).Conclusion:The osteogenic differentiation of type 2 diabetic rats mandibular marrow stromal cells was inhibited significantly. Morever,the decreased osteogenic differentiation ability cannot be restored even they were cultured in normal DMEM.