Effect of small molecules on dental pulp stem cells properties and pulp regeneration
Date
2015
DOI
Authors
Alhabib, Mey Abdulrahman
Version
OA Version
Citation
Abstract
BACKGROUND AND OBJECTIVES: Natural and synthetic small molecules have been shown to be useful chemical tools for controlling and manipulating the fates of cells. Here we show the effect of certain small molecules (Pluripotin (SC-1), GSK-3 inhibitor IX (BIO) and Rapamycin) on the stemness and differentiation potential of human dental pulp stem cells (hDPSCs) in vitro and how these molecules can target signaling transduction pathways and affect hDPSCs self-renewal and differentiation.
MATERIALS AND METHODS: Human dental pulps were extracted from teeth of healthy subjects aged 16-25 years. hDPSCs at passages 3 to 8 were first treated with various concentrations of the three chemicals to test their cell toxicity. Cells were then treated for five days with the final concentrations of 300 nM SC1, 100 nM BIO and 900 nM Rapamycin by choosing the concentration with the highest viable cells. Treated hDPSCs were analyzed for their proliferation, cell cycle profile, the expression levels of pluripotent and MSC markers, differentiation capacities (osteogenic, neurogenic & adipogenic), and intracellular signaling activations in comparison to the control nontreated hDPSC by immunocytostaining, flow cytometry analysis, RT-PCR, qPCR and western blot analysis.
RESULTS: Small molecule treatments decreased cell proliferation and enhanced the stemness of cultured hDPSCs evidenced by the increased expression of MSC and pluripotent stem cell markers STRO-1, NANOG, OCT4, and SOX2 and decreased expression of the senescent gene P16. Meanwhile, it diminished cell differentiation into odonto/osteogenic, adipogenic, and neurogenic lineages in vitro evidenced by decreased chemical staining (Alizarin red & Oil red staining) and differentiation gene expression. These effects involved Ras-GAP-, ERK1/2-, and mTOR-signaling pathways, which may preserve the cell self-renewal capacity, while suppressing differentiation.
CONCLUSION: We conclude that small molecules appear to enhance the immature state of hDPSCs in culture, which could be used as a strategy for adult stem cell maintenance and extend their capacity for regenerative applications.
Description
PLEASE NOTE: This work is protected by copyright. Downloading is restricted to the BU community: please click Download and log in with a valid BU account to access. If you are the author of this work and would like to make it publicly available, please contact open-help@bu.edu.
Dissertation (DScD) --Boston University, Henry M. Goldman School of Dental Medicine, 2015 (Department of Endodontics).
Includes bibliographic references: leaves 147-174.
Dissertation (DScD) --Boston University, Henry M. Goldman School of Dental Medicine, 2015 (Department of Endodontics).
Includes bibliographic references: leaves 147-174.
License
This work is protected by copyright. Downloading is restricted to the BU community. If you are the author of this work and would like to make it publicly available, please contact open-help@bu.edu.