Osteogenic differentiation potential of adipose-derived stem cells from ovariectomized mice.
Osteogenic differentiation potential of adipose-derived stem cells from ovariectomized mice.
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卵巢切除小鼠脂肪干细胞的成骨分化潜力
DOI：
10.1111/cpr.12328
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发表时间：
2017
影响因子：
8.5
通讯作者：
Xiao Jingang
中科院分区：
文献类型：
--
作者：
Wang Lei;Huang Chenglong;Li Qing;Xu Xiaomei;Liu Lin;Huang Kui;Cai Xiaoxiao;Xiao Jingang
骨质疏松症（OP）是一种由骨吸收和骨形成失衡引起的全身性疾病，通常由绝经后雌激素缺乏引起。虽然脂肪源性干细胞（ASCs）的成骨分化潜力已被证实，但OP对ASCs成骨分化的影响尚不清楚。在这里，我们的工作旨在比较从骨质疏松小鼠和正常对照小鼠获得的ASCs的增殖能力和成骨分化能力。材料与方法选取14周龄雌性C57BL/6小鼠20只，随机分为卵巢切除组（OVX）和假手术组（sham）两组（n=10）。ASCs和OP‐ASCs从女性腹股沟部位的皮下脂肪中获得。在随后的实验之前，细胞传代三次。使用xCELLigence系统监测细胞粘附和增殖。成骨诱导后，用茜素红染色分析分化ASCs和OP‐ASCs的矿化结节。real - time PCR检测骨桥蛋白（Opn）和runt相关转录因子2 （Runx2）等成骨特异性基因的表达，Western blotting检测骨相关蛋白的表达。结果6 d内各组细胞数量均稳定增加；48小时后，Sham组细胞增殖率明显高于OVX组。第21天，Sham组矿化骨结节结构明显高于OVX组，OVX组runx2 mRNA水平明显低于Sham组。编码opn的基因转录水平显示出与runx2相似的模式。Western blot结果显示，OVX组各时间点OPN、RUNX2蛋白表达水平均低于Sham组。结论与正常小鼠相比，骨质疏松小鼠ASCs的增殖能力和成骨潜能明显受损。然而，采用改良的OP‐ASCs自体移植治疗OP，或复合支架与改良的OP‐ASCs联合修复骨质疏松性骨缺损，可以克服其他方法的不足。
ObjectivesOsteoporosis (OP) is a systemic disease caused by imbalance between bone resorption and bone formation, commonly resulting from post‐menopausal oestrogen deficiency. Although osteogenic differentiation potential of adipose‐derived stem cells (ASCs) has been demonstrated, the effect of OP on osteogenic differentiation of ASCs remains unclear. Here, our work has been designed to compare proliferative capacity and osteogenic differentiation ability of ASCs obtained from osteoporotic mice and normal control mice.Materials and methodsTwenty 14‐week‐old female C57BL/6 mice were randomly divided into two groups: one, the ovariectomy (OVX) group (n=10), the other being the sham operated (Sham) group (n=10). ASCs and OP‐ASCs were obtained from subcutaneous fat of female inguinal sites. Cells were passaged three times prior to subsequent experimentation. The xCELLigence system was used to monitor cell adhesion and proliferation. Mineralized nodules of differentiated ASCs and OP‐ASCs were analysed using Alizarin red staining after osteogenic induction. Expressions of osteogenic‐specific genes including osteopontin (Opn) and runt‐related transcription factor 2 (Runx2) were assessed by real‐time PCR and expression of bone‐related proteins was detected by Western blotting.ResultsNumbers of cells in all groups increased steadily for 6 days; rate of cell proliferation in the Sham group was found to be higher than in the OVX group after 48 hours. Mineralized bone nodular structures were significantly more concentrated in the Sham group than in the OVX group by day 21, and mRNA levels ofRunx2in the OVX group were significantly lower than in the Sham group. Transcript levels of genes coding forOpnshowed a similar pattern to those ofRunx2. Western blot results indicated that protein expression levels of OPN and RUNX2 in the OVX group were lower than those in the Sham group, at each time point.ConclusionsThese results indicated that the proliferative capacity and osteogenic potential of ASCs were significantly impaired in osteoporotic mice compared to normal controls. However, use of autologous transplantation of modified OP‐ASCs for treatment of OP, or combination of composite scaffolds and modified OP‐ASCs for repair of osteoporotic bone defects, can overcome shortcomings of other methods.