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Adenovirus-mediated bone morphogenetic protein-2 promotes osteogenic differentiation in human mesenchymal stem cells in vitro

机译:腺病毒介导的骨形态发生蛋白-2在体外促进人间充质干细胞中的骨质发生分化

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摘要

Delayed and failed bone union following fracture is a common clinical complication that requires treatment in orthopedics. Cell-based therapies and tissue-engineering approaches are potential therapeutic strategies for bone repair and fracture healing. However, the effect of adenovirus expressing bone morphogenetic protein-2 (Ad-BMP-2) on the osteogenic ability of human mesenchymal stem cells (hMSCs) has remained to be fully elucidated. Therefore, in the present study, hMSCs were transduced using Ad-BMP-2 to assess the effects of its application and to determine whether Ad-BMP-2 promotes the osteogenic differentiation of hMSCs. The purity of the hMSC cultures was assessed using flow cytometric analysis. In order to assess the osteogenic activity, alkaline phosphatase activity (ALP) was measured and to estimate the osteoblastic mineralization and calcification, von Kossa staining for phosphates was performed. Cells positive for Src homology 2 domain were deterniined to be hMSCs and the presence of CD34 was used to distinguish hematopoietic lineages. Following treatment, the Ad-BMP-2 and control group had significantly increased ALP levels (P<0.05). Compared to the blank group and the group transfected with adenoviral vector containing LacZ, the phosphate deposition in the Ad-BMP-2 group and the positive control group treated with dexamethasone was markedly increased. The results of the present study suggested that Ad-BMP-2 promotes osteogenic differentiation in hMSCs and may have a potential application in treating delayed union and nonunion following bone fracture.
机译:骨折后延迟和失败的骨联盟是一种常见的临床并发症,需要在整形外科治疗。基于细胞的疗法和组织 - 工程方法是骨修复和骨折愈合的潜在治疗策略。然而,腺病毒表达骨形态发生蛋白-2(Ad-BMP-2)对人间充质干细胞(HMSCs)的成骨能力的影响保持完全阐明。因此,在本研究中,使用Ad-BMP-2转导HMSC以评估其应用的效果并确定Ad-BMP-2是否促进HMSCs的骨质发生分化。使用流式细胞术分析评估HMSC培养物的纯度。为了评估成骨活性,测量碱性磷酸酶活性(ALP)并估计骨细胞矿化和钙化,进行磷酸盐的vonKossa染色。 SRC同源性2结构域阳性的细胞呈HMSCs,并且使用CD34的存在来区分造血谱系。治疗后,AD-BMP-2和对照组的ALP水平显着增加(P <0.05)。与空白组和用含有LacZ的腺病毒载体转染的组相比,Ad-BMP-2组中的磷酸盐沉积和用地塞米松处理的阳性对照组被显着增加。本研究的结果表明,Ad-BMP-2促进HMSC中的骨质发生分化,并且可能具有在骨折后处理延迟联合和非同源的潜在应用。

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