DMP1-mediated transformation of DPSCs to CD31<sup>+</sup>/CD144<sup>+</sup> cells demonstrate endothelial phenotype both <i>in vitro</i> and <i>in vivo</i>.

Abstract

<h4>Introduction</h4>Dental pulp stem cells (DPSCs), can differentiate into endothelial cells (ECs), offering a promising strategy for generation of new blood vessels which is crucial for tissue repair and regeneration. Many studies have focused on optimizing conditions for differentiating DPSCs into ECs <i>in vitro</i> and subsequent validation of the vasculogenic potential of newly generated ECs <i>in vivo</i>. Previously, we demonstrated the ability of the HUVEC ECM scaffold along with DMP1 stimulation would drive endothelial-specific lineage of DPSCs.<h4>Methods</h4>In this study, DMP1-treated DPSCs were cultured on HUVEC ECM for 7 days and sorted using angiogenic-specific markers CD31 and CD144. The cells were separated into a positive fraction (CD31⁺/CD144⁺) and a negative fraction (CD31^-/CD144^-). To assess if ECs transformed from DMP1 stimulated DPSCs maintain their endothelial properties over time, we cultured both the positive CD31⁺/CD144⁺ and negative CD31^-/CD144^- fractions along with unstimulated DPSCs and assessed their angiogenic characteristics by gene expression analysis, functional properties using a tubule formation assay and <i>in vivo</i> subcutaneous implantation model.<h4>Results and discussion</h4>The findings of this study indicate that the CD31⁺/CD144⁺ fraction, retains both the phenotypic and functional characteristics of ECs, in contrast to the CD31^-/CD144^- fraction. Furthermore, <i>in vivo</i> analysis of the sorted ECs using the subcutaneous implantation model exhibited neovascularization along with the expression of vasculogenic markers. Overall, DPSC-derived ECs obtained by stimulation with DMP1 and cultured on HUVEC-ECM function as typical vascular ECs. This strategy, could be exploited for the development of vasculogeneis and as a therapeutic potential for tissue repair and regeneration.