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.
Highlights
Regenerative medicine is an interdisciplinary and rapidly developing field of industry and medicine since the last decades [1]
A significant number of adult mesenchymal stem cells with various differentiation and proliferation potential derive from the oral cavity [6,7]. These stem cells are mainly toothrelated cells derived from different tissues, such as dental pulp stem cells (DPSCs), stem cells from apical papilla (SCAPs), periodontal ligament stem cells (PDLSCs), dental follicle progenitor cells (DFPCs), tooth germ stem cells (TGSCs), gingiva-derived mesenchymal stem cells (GMSCs), alveolar bone marrow mesenchymal stem cells (ABMSCs), and stem cells from human exfoliated deciduous teeth (SHEDs)
The Bone morphogenetic protein 7 (BMP-7) expression in the cell lysates of the DPSC-BMP7 cell line, cultured in control medium (CM), CM+, Osteoinductive medium (OM), and OM+ was examined by western blot analysis, and the BMP-7 content of the cell supernatants was investigated by BMP-7 ELISA
Summary
Regenerative medicine is an interdisciplinary and rapidly developing field of industry and medicine since the last decades [1]. A significant number of adult mesenchymal stem cells with various differentiation and proliferation potential derive from the oral cavity [6,7]. The simplicity of tissue harvest, the high proliferative activity, differentiation capabilities, and immunomodulatory properties of these cells offer several opportunities for application in regenerative medicine. Most of these cells were already proved to be useful for the regeneration of bone and other tissue types [8,9,10,11,12]
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