In Vitro Reparative Dentin: a Biochemical and Morphological Study |
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| Authors: | G Teti V Salvatore A Ruggeri L Manzoli M Gesi G Orsini M Falconi |
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| Institution: | 1. DIBINEM – Department of Biomedical and Neuromotor Sciences, University of Bologna;2. Department of Translational Research and New Technologies in Medicine and Surgery, University of Pisa;3. Department of Clinical Sciences and Stomatology, Polytechnic University of Marche, Ancona, Italy |
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| Abstract: | In this study, starting from human dental pulp cells cultured in vitro, we simulated reparative dentinogenesis using a medium supplemented with different odontogenic inductors. The differentiation of dental pulp cells in odontoblast-like cells was evaluated by means of staining, and ultramorphological, biochemical and biomolecular methods. Alizarin red staining showed mineral deposition while transmission electron microscopy revealed a synthesis of extracellular matrix fibers during the differentiation process. Biochemical assays demonstrated that the differentiated phenotype expressed odontoblast markers, such as Dentin Matrix Protein 1 (DMP1) and Dentin Sialoprotein (DSP), as well as type I collagen. Quantitative data regarding the mRNA expression of DMP1, DSP and type I collagen were obtained by Real Time PCR. Immunofluorescence data demonstrated the various localizations of DSP and DMP1 during odontoblast differentiation. Based on our results, we obtained odontoblast-like cells which simulated the reparative dentin processes in order to better investigate the mechanism of odontoblast differentiation, and dentin extracellular matrix deposition and mineralization.Key words: dental tissue, in vitro differentiation, DMP1, DSP, type I collagen |
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