Dental pulp stem cells DPSCs also known as dental

Dental pulp stem EZ Cap Reagent GG Supplier (DPSCs), also known as dental pulp-derived mesenchymal stem cells, are a multipotent adult stem cell population that has a high proliferative potential. DPSCs in this study isolated from adult teeth are easily accessible and cryopreservable for long periods (Papaccio et al., 2006; Laino et al., 2006). DPSCs were used as a model of a mineralizing system, in vitro and in vivo, to study the effects of EtOH. It has been shown in numerous studies that under various conditions, dental pulp stem cells can be induced toward odontogenic and osteogenic lineages. One study demonstrated that vascular endothelial growth factor (VEGF) gene could promote odontogenic differentiation in DPSCs in vitro (Zhang et al., 2014) while another identified that DPSCs undergo osteogenic differentiation through the NF-kB signaling pathway (Wang et al., 2013). DPSCs had the ability to differentiate toward both odontogenic and osteogenic lineages in the presence of a carboxymethyl cellulose-hydroxyapatite hybrid hydrogel (Teti et al., 2015). Furthermore, medium modification with bone morphogenetic protein 2 was shown to stimulate odontogenic differentiation and formation of an osteo-dentin matrix (Atalayin et al., 2016). Although DPSCs have long been studied for their regenerative capabilities in both dentistry and orthopedics, the molecular mechanisms controlling their stem cell potency have yet to be discovered. It has been shown that KDM6B, a lysine-specific demethylase, plays a key role in osteogenic differentiation by removing H3K27me3 from the promoters of osteogenic genes in human bone marrow stromal cells (BMSCs) (Ye et al., 2012). It has also identified KDM6B in controlling HOX expression through the removal of H3K27me3 in human BMSCs (Ye et al., 2012). A recent study has shown KDM6B to play a critical role in the epigenetic regulation of odontogenic differentiation in human DPSCs (Xu et al., 2013). In DPSCs, KDM6B knockdown studies resulted in decreased alkaline phosphatase activity and Alizarin Red staining, and reduced expression levels of marker genes, including osterix (OSX), osteocalcin (OCN), and osteopontin (OPN) (Xu et al., 2013). Decreased levels of these markers and mineralization parallel effects observed in postmenopausal osteoporotic patients, who exhibit significant reductions in the number of osteocytes and osteoblasts, and demineralization of compact and cancellous bone (Pavel et al., 2016). While DPSCs primarily differentiate to dentin and BMSCs differentiate to bone, both dentin and bone formation share similar mineralization matrix components. Using the DPSC model of mineralization will give us insight on the molecular effects of EtOH on mineralization matrix that is similar to the bone mineralization model of BMSCs. In this study, we explore the epigenetic effects of alcohol on DPSCs and a possible link between alcohol and mineralization through the dysregulation of KDM6B.
Materials and methods
Results
Discussion Alcohol abuse appears to lead to periodontal disease, tooth decay and mouth sores that are potentially precancerous (Tezal et al., 2001; Amaral Cda et al., 2008; Yusko et al., 2008; Park et al., 2014). Persons who abuse alcohol are at high risk of having seriously deteriorated teeth, gums and compromised oral health in general (Tezal et al., 2001; Amaral Cda et al., 2008; Yusko et al., 2008; Park et al., 2014). It is generally believed that alcohol may have toxic effects on various cellular functions, but we lack detailed information about molecular and cellular effects of alcohol on stem cell maintenance and the differentiation process. Our recent publication demonstrated that EtOH exposure induced significant transcriptomic alterations in human embryonic stem cells (hESCs) through DNA methylomic deregulation (Khalid et al., 2014a). Several studies have demonstrated effects of EtOH on DNA methylation, resulting in genetic and phenotypic changes (Pandey et al., 2008; Oberlander et al., 2008; Shukla et al., 2008; Ouko et al., 2009; Haycock, 2009; Liu et al., 2009; Moonat et al., 2010; Qiang et al., 2010; Miranda et al., 2010; Kaminen-Ahola et al., 2010). It has been shown that EtOH induced alterations in DNA methylation patterns and inhibited neural stem cell (NSC) differentiation (Zhou et al., 2011). EtOH induced the hypermethylation of multiple cell cycle genes and increased the expression of DNA methyltransferases in NSCs. These alterations affected growth factor signaling, in conjunction with the down regulation of associated mRNAs and cell cycle proteins (Hicks et al., 2010). In another study, EtOH exposure prevented the methylation of specific genes related to neural development, including insulin-like growth factor 1 (IGF1), epidermal growth factor-containing fibulin-like extracellular matrix protein (EFEMP1), and SRY-box-containing gene 7 (SOX7) (Zhou et al., 2011). The hyper- or hypomethylation of specific genes have been shown to significantly affect NSCs\' differentiation and proliferation. However, it is important to note that many studies related to EtOH exposure and adult stem cell potency have shown that the intrinsic genetic and epigenetic mechanisms that control cellular fate are potentially of equal significance.