Evaluation of 5-Azacytidine as an inducer of osteoblastic / cementoblastic differentiation of progenitor cells from the periodontal ligament of humans

Resumo

Periodontal ligament (PDL) is constituted by heterogeneous cell populations with different cell differentiation capacities, here we investigated the effect of an epigenetic modulator on the induction of osteo/cementoblastic differentiation of human periodontal ligament mesenchymal progenitor cell clones with positive surface markers for CD105 (hPDLCD105+) and with low osteo/cementoblastic potential (LOP). These clones were pretreated with 5-aza-2'-deoxycytidine (5-aza-dC) and subjected to mineralization assays (Alizarin Red and Xylenol Orange), transcript levels’ analysis of ALP, ANXA2, ASPN, BMP2, FGF7, OCN, OPN and RUNX2, using qRT-PCR. As results, 5-aza-dC enhances the capacity of mineralized matrix deposition of two LOP clones (L1 and L2), however different results regarding to the quantity of calcium nodules production was found, where L1 mineralization and osteogenic-related gene expression was greater than L2. The data show that the augmentation of BMP2, FGF7 and ANXA2 transcript levels may lead to a better osteo/cementogenic response, while the elevation of ASPN gene expression may block the mineralization performance of LOP clones. These results showed that pre-treatment with 5-aza-dC can positively modulate the osteo/cementogenic differentiation of hPDL-CD105+ cell clones with low osteo/ emento-blastic potential. Those findings contribute to better understand the complex mechanisms that can lead PDL stem cells to regenerate the periodontal tissues.