Genome-wide DNA-methylation Profiles in Human Bone Marrow Mesenchymal Stem Cells on Titanium Surfaces

The characteristics of titanium (Ti) have been shown to influence dental implant fixation. Treatment of surfaces using the sandblasted, large-grit, acid-etched (SLA) method is widely used to provide effective osseointegration. However, the DNA methylation-associated mechanism by which SLA surface treatment affects osseointegration of human bone marrow mesenchymal stem cells (hBMSCs) remains elusive. Genome-wide methylation profiling of hBMSCs on SLA-treated and machined smooth Ti was performed using Illumina Infinium Methylation EPIC BeadChip at day 7 of osteogenic induction. In total, 2,846 CpG sites were differentially methylated in the SLA group compared with the machined group. Of these sites, 1,651 (covering 1,066 genes) were significantly hypermethylated and 1,195 (covering 775 genes) were significantly hypomethylated. Thirty significant enrichment pathways were observed, with Wnt signaling being the most significant. mRNA expression was identified by microarray and combined with DNA-methylation profiles. Thirty-seven genes displayed negative association between mRNA expression and DNA-methylation level, with the osteogenesis-related genes insulin-like growth factor 2 (IGF2) and carboxypeptidase X, M14 Family Member 2 (CPXM2) showing significant up-regulation and down-regulation, respectively. In summary, our results demonstrate differences between SLA-treated and machined surfaces in their effects on genome-wide DNA methylation and enrichment of osteogenic pathways in hBMSCs. We provide novel insights into genes and pathways affected by SLA treatment in hBMSCs at the molecular level.