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慢病毒介导的SATB2表达促进骨髓基质细胞在体外和体内的成骨分化。

Lentiviral-mediated expression of SATB2 promotes osteogenic differentiation of bone marrow stromal cells in vitro and in vivo.

作者信息

Gong Yiming, Qian Yanyan, Yang Fei, Wang Huijun, Yu Youcheng

机构信息

Department of Stomatology, Zhongshan Hospital, Fudan University, Shanghai, China.

出版信息

Eur J Oral Sci. 2014 Jun;122(3):190-7. doi: 10.1111/eos.12122. Epub 2014 Mar 26.

Abstract

Special AT-rich sequence-binding protein 2 (SATB2 ) acts as a potent transcription factor to promote osteoblast differentiation and bone regeneration. In this study, we first used lentiviral-mediated gene transfer of Satb2 into mouse bone marrow stromal cells (BMSCs) and investigated the capacity of SATB2 overexpression to promote osteogenic differentiation in vitro and in vivo. We found that LV-Satb2 -transduced BMSCs produced SATB2 protein and underwent rapid and marked osteogenic differentiation, as demonstrated by increased expression of osteoblastic genes, including runt-related transcription factor 2 (Runx2), transcription factor Sp7 (Sp7), activating transcription factor 4 (Atf4), and bone sialoprotein (Bsp), and increased alkaline phosphatase activity and Alizarin Red S staining. To analyze the induction of bone formation in vivo, LV-Satb2-transduced BMSCs were implanted into the hindlimbs of syngeneic mice, with β-tricalcium phosphate as the scaffolding material. Four weeks after implantation, transduction with LV-Satb2 had greatly enhanced the formation of new bone. These data demonstrated the capacity of lentiviral-mediated SATB2 to promote the osteogenic differentiation of BMSCs in vitro and to enhance bone formation through a tissue-engineering technique that may be useful in bone-regenerative medicine.

摘要

富含AT序列的特殊结合蛋白2(SATB2)作为一种有效的转录因子,可促进成骨细胞分化和骨再生。在本研究中,我们首先通过慢病毒介导的基因转移将Satb2导入小鼠骨髓间充质干细胞(BMSC),并研究SATB2过表达在体外和体内促进成骨分化的能力。我们发现,经LV-Satb2转导的BMSC产生SATB2蛋白,并迅速且显著地发生成骨分化,这表现为成骨细胞基因表达增加,包括与 runt 相关的转录因子2(Runx2)、转录因子Sp7(Sp7)、激活转录因子4(Atf4)和骨唾液蛋白(Bsp),以及碱性磷酸酶活性和茜素红S染色增加。为了分析体内骨形成的诱导情况,将经LV-Satb2转导的BMSC植入同基因小鼠的后肢,以β-磷酸三钙作为支架材料。植入四周后,LV-Satb2转导极大地促进了新骨形成。这些数据证明了慢病毒介导的SATB2在体外促进BMSC成骨分化以及通过组织工程技术增强骨形成的能力,这可能对骨再生医学有用。

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