The proangiogenic effects of extracellular vesicles secreted by dental pulp stem cells derived from periodontally compromised teeth

Abstract

Background: Although dental pulp stem cells (DPSCs) isolated from periodontally compromised teeth (P-DPSCs) have been demonstrated to retain pluripotency and regenerative potential, their use as therapeutics remains largely unexplored. In this study, we investigated the proangiogenic effects of extracellular vesicles (EVs) secreted by P-DPSCs using in vitro and in vivo testing models. Methods: Patient-matched DPSCs derived from periodontally healthy teeth (H-DPSCs) were used as the control for P-DPSCs. Conditioned media (CMs) derived from H-DPSCs and P-DPSCs (H-CM and P-CM), CMs derived from both cell types pretreated with the EV secretion blocker GW4869 (H-GW and P-GW), and EVs secreted by H-DPSCs and P-DPSCs (H-EVs and P-EVs) were prepared to test their proangiogenic effects on endothelial cells (ECs). Cell proliferation, migration, and tube formation were assessed using the Cell Counting Kit-8 (CCK-8), transwell/scratch wound healing, and Matrigel assays, respectively. Specifically, quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) and western blot analysis were used to examine the expression levels of angiogenesis-related genes/proteins in ECs in response to EV-based incubation. Finally, a full-thickness skin defect model was applied to test the effects of EVs on wound healing and new vessel formation.

Exosomes From LPS-stimulated hDPSCs Promote the Angiogenesis of HUVECs

“…In another study, when cocultured with exosomes derived from LPSpretreated hDPSCs, Schwann cells showed better migration and odontoblast differentiation abilities [28]. Furthermore, EVs from periodontitis-hDPSCs exhibited a stronger effect on angiogenesis and wound healing [13]. In our study, we demonstrated that the stronger proangiogenic paracrine activity of in ammation-induced hDPSCs was mediated by exosomes.…”

“…Interestingly, EVs secreted by in ammatory hDPSCs showed superior abilities in new vessel formation and cutaneous wound healing compared to EVs secreted by healthy teeth. Taken together, these results raised the question of whether exosomes from in ammatory hDPSCs contribute to improved angiogenic abilities [13]. In this study, we hypothesized that exosomes derived from hDPSCs from the in ammatory environment have stronger proangiogenesis effects, and these properties are mediated by speci c exosomal microRNAs.…”

Angiogenesis in Regenerative Dentistry: Are We Far Enough for Therapy?

“…The latest research direction is the use of exosomes. Zhou et al proved that exosomes derived from DPSCs of periodontally compromised teeth, possess proangiogenic effects, meaning that they might promote vascularization in regenerative endodontic therapy [ 129 ]. He also pointed out that exosomes isolated from cells under an inflammation status, present a much higher proangiogenic potential.…”

Bone Marrow Mesenchymal Stem Cell-Derived Exosomal miR-25 Regulates the Ubiquitination and Degradation of Runx2 by SMURF1 to Promote Fracture Healing in Mice

“…Totally, 2 × 10 6 MC3T3-E1C cells were seeded in the basolateral chambers of Transwell (3412, Corning, USA) while 4 × 10 5 MSCs or MSCs treated with 10 µM GW4869 (inhibitor for exosome secretion) (D1692, Sigma, USA) were plated in the apical chambers (13,14). After culture for 4-5 days, medium was renewed every 1-2 days.…”