Lithium-Activated SHEDs For Vascularized Dental Pulp Regeneration

This study aimed to investigate the effects of lithium chloride (LiCl) on the pro-angiogenic capacity and odontogenic differentiation of stem cells from human exfoliated deciduous teeth (SHEDs), and to evaluate their efficacy for dental pulp regeneration. SHEDs were treated with LiCl in vitro . Expression of angiogenic factors was evaluated by qRT-PCR and western blot. SHEDs-conditioned medium was collected to examine its effects on endothelial cell migration and tube formation via Transwell and Matrigel assays. Odontogenic differentiation was assessed using qRT-PCR, western blot, immunofluorescence, Alizarin Red staining, and ALP staining. For in vivo studies, LiCl and SHEDs were co-loaded into F127 hydrogel as a biocompatible scaffold, implanted into root segments, and transplanted subcutaneously into nude mice for 4 weeks. Tissue regeneration was analyzed by HE staining, CD31 immunofluorescence, and DSPP immunohistochemistry. LiCl treatment significantly enhanced angiogenic factor secretion by SHEDs, promoting endothelial cell migration and tube formation. Odontogenic differentiation and mineralization capacity of SHEDs were notably increased after LiCl stimulation. In vivo , SHEDs and LiCl co-delivered via F127 hydrogel regenerated vascularized pulp-like tissue with abundant CD31-positive vessels and DSPP-positive odontoblast-like cells aligning along dentin walls. Lithium chloride enhances the pro-angiogenic activity and odontogenic differentiation of SHEDs, and F127 hydrogel-mediated co-delivery promotes successful regeneration of vascularized dental pulp tissue, presenting a promising strategy for dental pulp regeneration.