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诱导型骨形态发生蛋白 7 表达对人牙髓干细胞成骨分化的影响。

Effect of Inducible BMP-7 Expression on the Osteogenic Differentiation of Human Dental Pulp Stem Cells.

机构信息

Department of Biomaterials and Prosthetic Dentistry, Faculty of Dentistry, University of Debrecen, 4032 Debrecen, Hungary.

Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Debrecen, 4032 Debrecen, Hungary.

出版信息

Int J Mol Sci. 2021 Jun 8;22(12):6182. doi: 10.3390/ijms22126182.

Abstract

BMP-7 has shown inductive potential for in vitro osteogenic differentiation of mesenchymal stem cells, which are an ideal resource for regenerative medicine. Externally applied, recombinant BMP-7 was able to induce the osteogenic differentiation of DPSCs but based on our previous results with BMP-2, we aimed to study the effect of the tetracyclin-inducible BMP-7 expression on these cells. DPSC, mock, and DPSC-BMP-7 cell lines were cultured in the presence or absence of doxycycline, then alkaline phosphatase (ALP) activity, mineralization, and mRNA levels of different osteogenic marker genes were measured. In the DPSC-BMP-7 cell line, the level of BMP-7 mRNA significantly increased in the media supplemented with doxycycline, however, the expression of Runx2 and noggin genes was upregulated only after 21 days of incubation in the osteogenic medium with doxycycline. Moreover, while the examination of ALP activity showed reduced activity in the control medium containing doxycycline, the accumulation of minerals remained unchanged in the cultures. We have found that the induced BMP-7 expression failed to induce osteogenic differentiation of DPSCs. We propose three different mechanisms that may worth investigating for the engineering of expression systems that can be used for the induction of differentiation of mesenchymal stem cells.

摘要

BMP-7 已显示出诱导间充质干细胞体外成骨分化的潜力,间充质干细胞是再生医学的理想资源。重组 BMP-7 经体外应用可诱导 DPSCs 的成骨分化,但基于我们之前对 BMP-2 的研究结果,我们旨在研究四环素诱导的 BMP-7 表达对这些细胞的影响。在存在或不存在强力霉素的情况下培养 DPSCs、模拟细胞和 DPSC-BMP-7 细胞系,然后测量碱性磷酸酶(ALP)活性、矿化和不同成骨标记基因的 mRNA 水平。在 DPSC-BMP-7 细胞系中,补充有强力霉素的培养基中 BMP-7 mRNA 的水平显著增加,然而,只有在用强力霉素孵育 21 天后,在成骨培养基中 Runx2 和 noggin 基因的表达才被上调。此外,尽管 ALP 活性的检测显示在含有强力霉素的对照培养基中活性降低,但培养物中的矿物质积累保持不变。我们发现,诱导的 BMP-7 表达未能诱导 DPSCs 的成骨分化。我们提出了三种不同的机制,这些机制可能值得进一步研究,以开发可用于诱导间充质干细胞分化的表达系统。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bca4/8229115/54da4359587c/ijms-22-06182-g001.jpg

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