Modulation of osteoblastic/cementoblastic potential of mesenchymal stem cell clones from periodontal ligament by epigenetic mechanism

Abstract

Cells derived from the periodontal ligament (PDL) have previously been reported to have stem cell-like characteristics and to play an important part to promote the regeneration of periodontal tissue destroyed by periodontal disease. However, these populations have been heterogeneous, and thus far no specific cellular phenotype has yet been established as being more capable in re-building damage tissues. Considering the cementum root neoformation an important step for complete periodontal regeneration, the aim of present study will be: 1) to determine gene expression profile of cell clones isolated from periodontal ligament mesenchymal stem cells and characterized as highly osteoblast/cementoblast PDL stem cells, and 2) to assess the effect of 5- Azacytidine (5-AZA) on the osteoblast/cementoblast differentiation of cell clones that do not have osteogenic potential. Cell clones identified as having osteoblast potential (C-O) or not (C-F, fibroblast profile) will culture under osteogenic conditions and gene expression for RUNX2, ALP, OC, BSP, OPN, GADD45b, PLAP-1/ASPN, AXNA2, PTEN, FGF7 IGFBP3 e BMP-2 will assessed by Real-Time PCR. Next, C-F cell clones will be submitted to osteogenic induction associated or not with 5-AZA to evaluate: a) mineral nodule formation by xylenol orange/alizarin red staining and RUNX2, ALP, OC, BSP and OPN gene expression, b) modulation of GADD45b, PLAP-1/ASPN, AXNA2, PTEN, FGF7, IGFBP3 and BMP-2 expression, and c) DNA methylation and hidroxymethilation of genes expressed by C-F clones treated with 5-AZA. (AU)