Article

Effects of Morphogen and Scaffold Porogen on the Differentiation of Dental Pulp Stem Cells

Department of Cariology, Restorative Sciences, Endodontics, University of Michigan School of Dentistry, Ann Arbor, MI, USA.
Journal of endodontics (Impact Factor: 2.79). 11/2010; 36(11):1805-11. DOI: 10.1016/j.joen.2010.08.031
Source: PubMed

ABSTRACT Dental pulp tissue engineering is an emerging field that can potentially have a major impact on oral health. However, the source of morphogens required for stem cell differentiation into odontoblasts and the scaffold characteristics that are more conducive to odontoblastic differentiation are still unclear. This study investigated the effect of dentin and scaffold porogen on the differentiation of human dental pulp stem cells (DPSCs) into odontoblasts.
Poly-L-lactic acid (PLLA) scaffolds were prepared in pulp chambers of extracted human third molars using salt crystals or gelatin spheres as porogen. DPSCs seeded in tooth slice/scaffolds or control scaffolds (without tooth slice) were either cultured in vitro or implanted subcutaneously in immunodefficient mice.
DPSCs seeded in tooth slice/scaffolds but not in control scaffolds expressed putative odontoblastic markers (DMP-1, DSPP, and MEPE) in vitro and in vivo. DPSCs seeded in tooth/slice scaffolds presented lower proliferation rates than in control scaffolds between 7 and 21 days (p < 0.05). DPSCs seeded in tooth slice/scaffolds and transplanted into mice generated a tissue with morphological characteristics similar to those of human dental pulps. Scaffolds generated with gelatin or salt porogen resulted in similar DPSC proliferation. The porogen type had a relatively modest impact on the expression of the markers of odontoblastic differentiation.
Collectively, this work shows that dentin-related morphogens are important for the differentiation of DPSC into odontoblasts and for the engineering of dental pulp-like tissues and suggest that environmental cues influence DPSC behavior and differentiation potential.

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    • "The interchange of responsive cells, morphogens, and scaffolds constitutes the three main elements that grounds tissue engineering [1] [2] [3] [4] [5] [6]. Scaffolds are three-dimensional structures used to support and guide the in-growth of cells, forming the template for cell colonization, proliferation as well as being able to provide different sets of physiological signals to the developing tissue [7] [8]. Therefore scaffolds perform the structural and biochemical functions of the native extracellular matrix (ECM) until the cells are able to produce their own ECM [9] [10]. "
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    • "The stem cells of dental origin can certainly generate the dental tissues [10] [11] [12] [13] [14]. The SHED and DPSCs are capable of generating a tissue that has morphological and functional characteristics that closely resemble those of the human dental pulp [15] [16] [17] [18]. "
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    • "Real-time PCR was used to compare dentin sialophosphoprotein (DSPP) and type I collagen gene expression, as markers of pulp cell differentiation [Bleicher et al., 1999; About et al., 2000; Domon et al., 2001; Liu et al., 2005; Zhang et al., 2005; Demarco et al., 2010]. Total RNA was isolated from the cultures using an RNeasy Mini Kit (Qiagen, Valencia, Calif., USA), and 2.5 ␮ g purified total RNA was reverse-transcribed into cDNA using the SuperScript III Reverse Transcriptase kit (Invitrogen). "
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