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Roles of L-type calcium channels (Ca V 1.2) and the distal C-terminus (DCT) in differentiation and mineralization of rat dental apical papilla stem cells (rSCAPs).
- Source :
-
Archives of oral biology [Arch Oral Biol] 2017 Feb; Vol. 74, pp. 75-81. Date of Electronic Publication: 2016 Nov 04. - Publication Year :
- 2017
-
Abstract
- Objective: Voltage-gated inward Ca <superscript>2+</superscript> currents (ICa) are triggered by cell depolarization and commonly produce transient increases in the cytoplasmic free Ca <superscript>2+</superscript> concentration. The Ca <subscript>V</subscript> 1.2 distal C-terminus is susceptible to proteolytic cleavage, which yields a truncated Ca <subscript>V</subscript> 1.2 subunit and a cleaved C-terminal fragment (CCt or DCT). Stem cells from the apical papilla (SCAPs) has a capacity for differentiation into the odontoblastic-like cells in vitro and dentin forming in vivo, which makes SCAPs advantages in tissue engineering and regenerative endodontic. The aim of this study was to investigate the effect of Ca <subscript>V</subscript> 1.2 and its distal C-terminal fragment in the odontoblastic differentiation of rat SCAPs (stem cells from the apical papilla).<br />Design: In this study, we generated stable Ca <subscript>V</subscript> 1.2 knockdown and DCT over-expressed rSCAPs using short hairpin RNA and DCT gene containing Lentivirus vectors, respectively. The transfected apical papilla cells were induced to differentiate into the odontoblast-like cells, and the expression of markers for odontoblastic differentiation were analyzed by alizarin red staining, Real-time Polymerase chain reaction (RT-PCR), and Western blot analysis.<br />Results: The knockdown of Ca <subscript>V</subscript> 1.2 and excess expression of DCT both suppressed the expression of DSPP, ALP in mRNA level and the formation of calcium nodules.<br />Conclusions: Our results suggest that Ca <subscript>V</subscript> 1.2 and DCT play important roles in the differentiation of rSCAPs, DCT might act as a transcription factor and regulate the differentiation of rSCAPs.<br /> (Copyright © 2016 Elsevier Ltd. All rights reserved.)
- Subjects :
- Animals
Calcium Channels, L-Type genetics
Cell Survival
Cells, Cultured
Dental Papilla cytology
Extracellular Matrix Proteins metabolism
Gene Expression Regulation
Gene Knockdown Techniques
Genetic Vectors genetics
Lentivirus genetics
Male
Phosphoproteins metabolism
RNA, Small Interfering
Rats
Rats, Sprague-Dawley
Sialoglycoproteins metabolism
Stem Cells cytology
Calcium Channels, L-Type metabolism
Calcium Channels, L-Type physiology
Cell Differentiation
Dental Papilla metabolism
Odontoblasts metabolism
Stem Cells metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1879-1506
- Volume :
- 74
- Database :
- MEDLINE
- Journal :
- Archives of oral biology
- Publication Type :
- Academic Journal
- Accession number :
- 27918898
- Full Text :
- https://doi.org/10.1016/j.archoralbio.2016.11.004