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腺病毒介导的骨形态发生蛋白-2促进人间充质干细胞的成骨分化。

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

作者信息

Cao Hong, Sun Zhi-Bo, Zhang Lei, Qian Wei, Li Chun-Yang, Guo Xiao-Peng, Zhang Ying

机构信息

Department of Orthopedic Surgery, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China.

Department of Reproductive Medicine, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei 442000, P.R. China.

出版信息

Exp Ther Med. 2017 Jul;14(1):377-382. doi: 10.3892/etm.2017.4482. Epub 2017 May 22.

Abstract

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 determined 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.

摘要

骨折后延迟骨愈合和骨不连是骨科临床常见的并发症,需要进行治疗。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,表达骨形态发生蛋白2的腺病毒(Ad-BMP-2)对人间充质干细胞(hMSCs)成骨能力的影响仍有待充分阐明。因此,在本研究中,使用Ad-BMP-2转导hMSCs,以评估其应用效果,并确定Ad-BMP-2是否促进hMSCs的成骨分化。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),并进行von Kossa磷酸盐染色以估计成骨细胞的矿化和钙化。确定具有Src同源2结构域阳性的细胞为hMSCs,并使用CD34的存在来区分造血谱系。治疗后,Ad-BMP-2组和对照组的ALP水平显著升高(P<0.05)。与空白组和转染含LacZ腺病毒载体的组相比,Ad-BMP-2组和用地塞米松治疗的阳性对照组的磷酸盐沉积明显增加。本研究结果表明,Ad-BMP-2促进hMSCs的成骨分化,可能在治疗骨折后延迟愈合和骨不连方面具有潜在应用价值。

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