Abstract
BackgroundDental pulp tissue contains many undifferentiated mesenchymal cells, which retain the ability to differentiate into mature cells. Induced pluripotent stem cells have been developed from various cell sources, including dental pulp-derived stem cells, and evaluated for potential application to regenerative therapy. Dental pulp tissues overexpress CD44, a cell-adhesion factor involved in the induction of mineralization. In this study, we investigated the effects of hyaluronan—a known CD44 ligand—on dental pulp stem cells (DPSCs).MethodsDPSC CD44 expression was analyzed using immunofluorescence staining, flow cytometry, and western blotting. Cell proliferation was evaluated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Effects of hyaluronan on the cell cycle were analyzed by flow cytometry. Alkaline phosphatase activity was employed as marker of mineralization and measured by fluorometric quantification and western blotting. Bone morphogenetic protein (BMP)-2, BMP-4, dentin sialophosphoprotein (DSPP), and dentin matrix acidic phosphoprotein 1 (DMP-1) levels were measured using real-time polymerase chain reaction. Odontoblastic differentiation and the close cell signaling examination of DPSC differentiation were determined using western blotting.ResultsHyaluronan induced expression of the odontoblastic differentiation markers DMP-1 and DSPP. Moreover, the odontoblastic differentiation induced by hyaluronan was mediated by CD44—but not by Akt, Smad1 or MAPK signaling.ConclusionsOur results indicate that hyaluronan induces odontoblastic differentiation of DPSCs via CD44. This suggests that hyaluronan plays a crucial role in the induction of odontoblastic differentiation from DPSCs. Our findings may aid the development of new, inexpensive, and effective conservative treatments for dental pulp repair.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-016-0399-8) contains supplementary material, which is available to authorized users.
Highlights
Dental pulp tissue contains many undifferentiated mesenchymal cells, which retain the ability to differentiate into mature cells
Expression of CD44 in dental pulp stem cells (DPSCs) Most CD44 antigenicity in dental pulp tissue is present in the incomplete region of the roots [12]
Odontoblastic differentiation of DPSCs induced by HA treatment involves CD44 signaling Because HA is a known ligand of CD44, we investigated whether HA-mediated dentin matrix acidic phosphoprotein 1 (DMP-1) expression and odontoblastic differentiation arose via CD44 signaling
Summary
Dental pulp tissue contains many undifferentiated mesenchymal cells, which retain the ability to differentiate into mature cells. Dental pulp stem cells (DPSCs) are present in human dental pulp, even in adult pulp, as clonogenic and highly proliferative cells obtained after enzymatic disaggregation [2]. These cells harbor the characteristics of plastic adherence and express stem cell markers such as CD29, CD90, CD44, and CD146 [2]. Numerous researchers have since shown that DPSCs retain the capacity for both self-renewal and multiple cell lineage differentiation [5, 6] and can be stimulated, under specific conditions, to differentiate into various cell types such as adipocytes, myoblasts, neurons, Umemura et al Stem Cell Research & Therapy (2016) 7:135 chondrocytes, odontoblasts and osteoblasts both in vitro and in vivo [7,8,9]. Animal studies have revealed great potential for DPSCs in the repair and regeneration of various tissues, including bone [10], muscle [3] and teeth [11]
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