Abstract

Odontoblasts, the main cell type in teeth pulp tissue, are not cultivable and they are responsible for the first line of response after dental restauration. Studies on dental materials cytotoxicity and odontoblast cells physiology require large quantity of homogenous cells retaining most of the phenotype characteristics. Odontoblast-like cells (OLC) were differentiated from human dental pulp stem cells using differentiation medium (containing TGF-β1), and OLC expanded after trypsinization (EXP-21) were evaluated and compared. Despite a slower cell growth curve, EXP-21 cells express similarly the odontoblast markers dentinal sialophosphoprotein and dentin matrix protein-1 concomitantly with RUNX2 transcripts and low alkaline phosphatase activity as expected. Both OLC and EXP-21 cells showed similar mineral deposition activity evidenced by alizarin red and von Kossa staining. These results pointed out minor changes in phenotype of subcultured EXP-21 regarding the primarily differentiated OLC, making the subcultivation of these cells a useful strategy to obtain odontoblasts for biocompatibility or cell physiology studies in dentistry.

Highlights

  • Odontoblasts are highly specialized cells that produce both collagen and noncollagen proteins to build the dentinal extracellular matrix

  • The genotype, phenotype, or cell responses in these in vitro models could differ from actual odontoblast cell responses due to genetic changes [8] or environmental adaptations of the cell lines, which complicates the interpretation of cytotoxicity results

  • The results showed that Human dental pulp stem cells (hDPSC) differentiate to odontoblast-like cells (OLC) in the presence of mineralizing medium enriched with TGFβ1

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Summary

Introduction

Odontoblasts are highly specialized cells that produce both collagen and noncollagen proteins to build the dentinal extracellular matrix They are the pulp first responders against exogenous stimulus or dental materials [1, 2]. Due to their postmitotic phenotype, odontoblasts are difficult to culture [3, 4]; biochemical or toxicologic studies assessing their use in dental materials have been limited. To overcome such difficulties, studies on materials toxicity have been developed on primary gingival fibroblasts [5], primary human or mouse undifferentiated mesenchymal stem cells, and immortalized cell lines [6, 7]. Differentiated odontoblasts establish a structure known as the dentin-pulp complex; these cells have been proposed as tools in dental regeneration and repair [9]

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