Modulation of osteoblastic-cementoblastic potencial of mesenchymal stem cell clones from periodontal ligament by epigenetic mechanism

Author(s)

Catharina Marques Sacramento

• Document type: Master's Dissertation
• Press: Piracicaba, SP.
• Institution: Universidade Estadual de Campinas (UNICAMP). Faculdade de Odontologia de Piracicaba
• Defense date: 2019-02-22

Examining board members

Karina Gonzales Silverio; Rodrigo Augusto da Silva; Thaisângela Rodrigues

• Advisor: Karina Gonzales Silverio; Denise Carleto Andia

Abstract

Periodontal ligament (PDL) is constituted by heterogeneous cell populations with different cell differentiation capacities. The mechanisms that regulate the involvement of mesenchymal progenitor cells of PDL with osteo/cementoblastic phenotype are not yet fully understood. 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 (hPDL-CD105+) and with low osteo/cementoblastic potential (LOP). These clones were pretreated with 5-aza-2'-deoxycytidine (5-aza-dC) e Dimethyl Sulfoxide (DMSO) for 48 hours 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 and gene-specific methylation/hydroxymethylation analysis of ASPN, BMP2 and FGF7 genes. 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. In addition, FGF7 methylation/hydroxymethylation high levels are correlated to low transcripts expression of this gene in the clone with poor response to 5-aza-dC (L2). Nevertheless, DMSO also enhances the osteo/cementogenic capacity of LOP clones. These results showed that pre-treatment with 5-aza-dC and DMSO 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 (AU)

• FAPESP's process: 17/19697-2 - Modulation of osteoblastic/cementoblastic potential of mesenchymal stem cell clones from periodontal ligament by epigenetic mechanism
• Grantee: Catharina Marques Sacramento
• Support Opportunities: Scholarships in Brazil - Master