Effects of Morphogen and Scaffold Porogen on the Differentiation of Dental Pulp Stem Cells
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.
- SourceAvailable from: Evandro Piva
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- "The interchange of responsive cells, morphogens, and scaffolds constitutes the three main elements that grounds tissue engineering      . 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  . Therefore scaffolds perform the structural and biochemical functions of the native extracellular matrix (ECM) until the cells are able to produce their own ECM  . "
ABSTRACT: We aimed to develop an alginate hydrogel (AH) modified with nano-/microfibers of titanium dioxide (nfTD) and hydroxyapatite (nfHY) and evaluated its biological and chemical properties. Nano-/microfibers of nfTD and nfHY were combined with AH, and its chemical properties were evaluated by FTIR spectroscopy, X-ray diffraction, energy dispersive X-Ray analysis, and the cytocompatibility by the WST-1 assay. The results demonstrate that the association of nfTD and nfHY nano-/microfibers to AH did not modified the chemical characteristics of the scaffold and that the association was not cytotoxic. In the first 3 h of culture with NIH/3T3 cells nfHY AH scaffolds showed a slight increase in cell viability when compared to AH alone or associated with nfTD. However, an increase in cell viability was observed in 24 h when nfTD was associated with AH scaffold. In conclusion our study demonstrates that the combination of nfHY and nfTD nano-/microfibers in AH scaffold maintains the chemical characteristics of alginate and that this association is cytocompatible. Additionally the combination of nfHY with AH favored cell viability in a short term, and the addition of nfTD increased cell viability in a long term.06/2013; 2013:307602. DOI:10.1155/2013/307602
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- "The stem cells of dental origin can certainly generate the dental tissues     . 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    . "
ABSTRACT: Stem cells are distinguished by their ability to differentiate into different types of cells in the body and to self-replicate. During the recent years, stem cells have been used extensively in the field of medicine for the repair and regeneration of defective tissues and organs. However, the knowledge on the stem cell technology is increasing quickly in all medical disciplines and it dictates the need for new protective approaches in all fields, which include reparative dentistry. Stem cell therapy constitutes a common challenge for dentists as well as for biotechnologists. The aim of this study was to review the knowledge which was related to stem cells and to consider the possibility of use of stem cell populations and their technology in the future clinical applications, to cure diseases like Parkinsonism, Juvenile diabetes, certain forms of cancer, spinal injuries and heart problems.05/2013; 7(5):945-7. DOI:10.7860/JCDR/2013/5379.2984
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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). "
ABSTRACT: Amelogenins containing exons 8 and 9 are alternatively spliced variants of amelogenin. Some amelogenin spliced variants have been found to promote pulp regeneration following pulp exposure. The function of the amelogenin spliced variants with the exons 8 and 9 remains unknown. In this study, we synthesized recombinant leucine rich amelogenin peptide (LRAP, A-4), LRAP plus exons 8 and 9 peptide (LRAP 8, 9) or exons 8 and 9 peptide (P89), to determine their effects on odontoblasts. In vivo analyses were completed following the insertion of agarose beads containing LRAP or LRAP 8, 9 into exposed cavity preparations of rat molars. After 8, 15 or 30 days' exposure, the pulp tissues were analyzed for changes in histomorphometry and cell proliferation by PCNA stainings. In vitro analyses included the effects of the addition of the recombinant proteins or peptide on cell proliferation, differentiation and adhesion of postnatal human dental pulp cells (DPCs). These studies showed that in vivo LRAP 8, 9 enhanced the reparative dentin formation as compared to LRAP. In vitro LRAP 8, 9 promoted DPC proliferation and differentiation to a greater extent than LRAP. These data suggest that amelogenin exons 8 and 9 may be useful in amelogenin-mediated pulp repair.Cells Tissues Organs 02/2012; 196(2):151-60. DOI:10.1159/000331248 · 2.14 Impact Factor