Growth and Osteogenic Properties of Dental Pulp Stem Cells from Deciduous vs. Permanent Teeth.
Abstract: ABSTRACT BACKGROUND: Mesenchymal stem cells (MSCs) isolated from dental pulp can differentiate into different cell types including osteoblasts and could therefore be used as a source for autologous bone engineered in vitro. In this study, using a 3D culture model, we compared the growth and osteogenic potential of MSCs isolated from human primary deciduous teeth to those isolated from hu... read moreman adult permanent teeth. MATERIAL AND METHODS: MSCs were isolated and propagated from six healthy permanent and six primary teeth. The colony forming potential (colony forming units (CFU) assay) of these cells was evaluated in non-osteogenic and osteogenic media. Dental pulp MSCs were also embedded in a scaffold material (hydrogel) and grown in osteogenic or non-osteogenic media for 4 weeks. Cell proliferation was evaluated by measuring total cellular DNA using a spectrofluorometric assay. Histology (hematoxylin and eosin) and the amount of mineralization (alizarin red) were evaluated on paraffin-embedded sections. Expression of the differentiation marker genes alkaline phosphatase, osteocalcin, dentin sialophosphoprotein, and bone sialoprotein was evaluated by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). RESULTS: Deciduous dental pulp MSCs formed significantly higher number of CFUs (p=0.0002) in DMEM and osteogenic media (p=0.0351) and proliferated at a higher rate in osteogenic media (P=0.0186) and DMEM (p=0.0052) when compared to adult dental pulp MSCs. Compared to adult, deciduous dental pulp MSCs cultured in hydrogel scaffolds showed a higher degree of mineralization and formed larger nodules in both 3 osteogenic (p=0.0478) and DMEM (p=0.0340). There were no significant differences in the level of expression of all evaluated genes between deciduous and adult dental pulp MSCs. Furthermore, neither cell type was able to completely hydrolyze the hydrogel scaffold. CONCLUSION: Deciduous dental pulp MSCs has a higher proliferation rate and mineralization potential compared to adult dental pulp MSCs making them an ideal candidate for tissue engineering. However, the hydrogel used in these studies does not seem to be an optimal scaffold for engineering bone-like structures.
Thesis (M.S.)--Tufts University, 2011.
Submitted to the Dept. of Pediatric Dentistry.
Advisor: Driss Zoukhri.
Committee: Paul Stark, Laura Cstro, Cheen Loo, and Pamela Yelcik.
Keywords: Dentistry, and Molecular biology.read less