中国组织工程研究
中國組織工程研究
중국조직공정연구
Journal of Clinical Rehabilitative Tissue Engineering Research
2013年
36期
6423-6429
,共7页
王云%鲍小明%侯永新%李军%张民
王雲%鮑小明%侯永新%李軍%張民
왕운%포소명%후영신%리군%장민
干细胞%骨髓干细胞%骨髓间充质干细胞%骨质疏松%绝经%去卵巢%成骨诱导%碱性磷酸酶%细胞周期%增殖指数%省级基金%干细胞图片文章
榦細胞%骨髓榦細胞%骨髓間充質榦細胞%骨質疏鬆%絕經%去卵巢%成骨誘導%堿性燐痠酶%細胞週期%增殖指數%省級基金%榦細胞圖片文章
간세포%골수간세포%골수간충질간세포%골질소송%절경%거란소%성골유도%감성린산매%세포주기%증식지수%성급기금%간세포도편문장
背景：细胞学研究表明，骨髓间充质干细胞在绝经后骨质疏松症的发病过程中起有重要作用。<br> 目的：观察去卵巢骨质疏松大鼠骨髓间充质干细胞的体外成骨分化。<br> 方法：将6月龄雌性SD大鼠双侧卵巢切除建立骨质疏松模型。实验分为4组：正常干细胞组、骨质疏松干细胞组、正常干细胞成骨诱导组、骨质疏松干细胞成骨诱导组。全骨髓贴壁法培养正常和骨质疏松大鼠骨髓间充质干细胞至第3代细胞用于实验。倒置相差显微镜观察细胞形态，流式细胞仪测定细胞周期、增殖指数。加成骨诱导液进行成骨诱导，检测各组骨髓间充质干细胞碱性磷酸酶活性，茜素红染色法比较各组钙结节的形成。<br> 结果与结论：正常干细胞成骨诱导组、骨质疏松干细胞成骨诱导组均具有成骨细胞的形态特征，但骨质疏松干细胞成骨诱导组形态变化相对缓慢。正常干细胞组细胞增殖指数高于骨质疏松干细胞组(P<0.05)；成骨诱导组碱性磷酸酶活性均明显高于相应的正常或骨质疏松干细胞组(P<0.05)；正常干细胞成骨诱导组明显高于骨质疏松干细胞成骨诱导组(P<0.05)。成骨诱导组茜素红染色均呈阳性，相应的正常或骨质疏松干细胞组呈阴性；且正常干细胞成骨诱导组染色强于骨质疏松干细胞成骨诱导组。提示去卵巢骨质疏松大鼠骨髓间充质干细胞的增殖能力和成骨分化能力明显降低，可能与去卵巢大鼠骨质疏松的发生相关。
揹景：細胞學研究錶明，骨髓間充質榦細胞在絕經後骨質疏鬆癥的髮病過程中起有重要作用。<br> 目的：觀察去卵巢骨質疏鬆大鼠骨髓間充質榦細胞的體外成骨分化。<br> 方法：將6月齡雌性SD大鼠雙側卵巢切除建立骨質疏鬆模型。實驗分為4組：正常榦細胞組、骨質疏鬆榦細胞組、正常榦細胞成骨誘導組、骨質疏鬆榦細胞成骨誘導組。全骨髓貼壁法培養正常和骨質疏鬆大鼠骨髓間充質榦細胞至第3代細胞用于實驗。倒置相差顯微鏡觀察細胞形態，流式細胞儀測定細胞週期、增殖指數。加成骨誘導液進行成骨誘導，檢測各組骨髓間充質榦細胞堿性燐痠酶活性，茜素紅染色法比較各組鈣結節的形成。<br> 結果與結論：正常榦細胞成骨誘導組、骨質疏鬆榦細胞成骨誘導組均具有成骨細胞的形態特徵，但骨質疏鬆榦細胞成骨誘導組形態變化相對緩慢。正常榦細胞組細胞增殖指數高于骨質疏鬆榦細胞組(P<0.05)；成骨誘導組堿性燐痠酶活性均明顯高于相應的正常或骨質疏鬆榦細胞組(P<0.05)；正常榦細胞成骨誘導組明顯高于骨質疏鬆榦細胞成骨誘導組(P<0.05)。成骨誘導組茜素紅染色均呈暘性，相應的正常或骨質疏鬆榦細胞組呈陰性；且正常榦細胞成骨誘導組染色彊于骨質疏鬆榦細胞成骨誘導組。提示去卵巢骨質疏鬆大鼠骨髓間充質榦細胞的增殖能力和成骨分化能力明顯降低，可能與去卵巢大鼠骨質疏鬆的髮生相關。
배경：세포학연구표명，골수간충질간세포재절경후골질소송증적발병과정중기유중요작용。<br> 목적：관찰거란소골질소송대서골수간충질간세포적체외성골분화。<br> 방법：장6월령자성SD대서쌍측란소절제건립골질소송모형。실험분위4조：정상간세포조、골질소송간세포조、정상간세포성골유도조、골질소송간세포성골유도조。전골수첩벽법배양정상화골질소송대서골수간충질간세포지제3대세포용우실험。도치상차현미경관찰세포형태，류식세포의측정세포주기、증식지수。가성골유도액진행성골유도，검측각조골수간충질간세포감성린산매활성，천소홍염색법비교각조개결절적형성。<br> 결과여결론：정상간세포성골유도조、골질소송간세포성골유도조균구유성골세포적형태특정，단골질소송간세포성골유도조형태변화상대완만。정상간세포조세포증식지수고우골질소송간세포조(P<0.05)；성골유도조감성린산매활성균명현고우상응적정상혹골질소송간세포조(P<0.05)；정상간세포성골유도조명현고우골질소송간세포성골유도조(P<0.05)。성골유도조천소홍염색균정양성，상응적정상혹골질소송간세포조정음성；차정상간세포성골유도조염색강우골질소송간세포성골유도조。제시거란소골질소송대서골수간충질간세포적증식능력화성골분화능력명현강저，가능여거란소대서골질소송적발생상관。
BACKGROUND:Cytological studies show that bone marrow mesenchymal stem cel s play an important role in postmenopausal osteoporosis mechanism. <br> OBJECTIVE:To study the osteogenic differentiation in vitro of bone marrow mesenchymal stem cel s from ovariectomied osteoporotic rats. <br> METHODS:The osteoporotic animal model was established by performing ovariectomy in the 6-month-old female Sprague-Dawley rats. There were four groups:bone marrow mesenchymal stem cel s control group, bone marrow mesenchymal stem cel s osteoporosis group, bone marrow mesenchymal stem cel s osteogenic induction group and oseogenesis induction group. Bone marrow mesenchymal stem cel s were isolated from the rats of control group and oseogenesis induction group by means of the whole bone marrow adherence method and cultured to the 3rd generation. Then the bone marrow mesenchymal stem cel s were used in al the experiments. Cel morphology was observed under the inverted phase contrast microscope, cel cycle and proliferation index of bone marrow mesenchymal stem cel s were detected by flow cytometry. After osteogenic induction, the expression level of alkaline phosphatase was detected, and the fornation of calcium nodes of bone marrow mesenchymal stem cel s were marked by alizarin red staining. <br> RESULTS AND CONCLUSION:The cel s in the osteogenic induction group and oseogenesis induction group had the morphology of osteobalsts, and the change of morphology of the cel s in the oseogenesis induction group was relatively tardiness. The proliferation index in the control group was higher than that in the osteoporosis group (P<0.05);expression level of alkaline phosphatase in the osteogenic induction group was significantly higher than that in the oseogenesis induction group (P<0. 05), and the control group was significantly higher than the oseogenesis group (P<0.05). The alizarin red staining of the cel s in the osteogenic induction group was positive, while negative in the control group and the oseogenesis group;the staining in the osteogenic induction group was stronger than that in the oseogenesis induction group. These findings indicate that both the proliferative potential and the osteogenic potential of bone marrow mesenchymal stem cel s from the ovariectomized osteoporotic rats are decreased, which may be related with the ostoeporosis pathogensis of ovariectomied rats.
