Pulp-Derived Exosomes in a Fibrin-Based Regenerative Root Filling Material

Abstract

Regenerative endodontics has been described as a paradigm shift in dentistry, despite its current limitation to immature teeth and reparative rather than regenerative outcomes. Cell-free treatments are favored because of regulatory issues. However, the recruitment of host-derived stem cells to the desired site remains challenging. We investigated whether dental pulp-derived exosomes, which are extracellular vesicles that contain proteins, lipids, RNA, and DNA and thus mirror their parental cells, may be used for this purpose. The use of exosomes may present appreciable advantages over the direct use of transplanted stem cells due to a higher safety profile, easier isolation, preservation, and handling. Here we harvested exosomes from a cultured third-molar pulp cell and assessed them by transmission electron microscopy and Western blotting. Human mesenchymal stem cells (MSCs) were exposed to these exosomes to assess exosome uptake, cell migration, and proliferation. In addition, a fibrin gel (i.e., a diluted fibrin sealant), was assessed as a delivery system for the exosomes. Our results show that exosomes attracted MSCs, and the fibrin gel enhanced their effect. Moreover, exosomes improved the proliferation of MSCs. Therefore, we propose that pulp-derived exosomes in combination with a fibrin gel could be a powerful combination for clinical translation towards improved cell-free regenerative endodontics and thus represent a new way to fill dental hard tissues.

The Case of Medication-Related Osteonecrosis of the Jaw Addressed from a Pathogenic Point of View. Innovative Therapeutic Strategies: Focus on the Most Recent Discoveries on Oral Mesenchymal Stem Cell-Derived Exosomes

“…Moreover, the exosomes derived from cells cultured in the presence of odontogenic differentiation media were more potent in inducing odontogenic differentiation, most probably as consequence of altered genetic and protein exosomal cargo ( Figure 3B) [113]. DPSCs-derived exosomes also induced BMSCs migration and proliferation [120]. Similar results were confirmed by Hu et al, 2019.…”

“…To date, a limited number of papers have described the isolation, composition and function of DPSCs exosomes [112][113][114][115][116][117][118][119][120]. Recently, a proangiogenic action for DPSCs-derived exosomes has been demonstrated.…”

Extracellular vesicles in bone and periodontal regeneration: current and potential therapeutic applications

“…Furthermore, the therapeutic use of EVs has also gained attention in reconstructing the pulp complex and dentin in recent years. Ivica et al [106] proposed that pulp-derived EVs, along with a fibrin gel, could be an effective combination for clinical translation on the way to improved cell-free regenerative endodontics. Furthermore, Huang et al assessed the potential characteristics of EVs from dental pulp stem cells that were cultured in odontogenic differentiation conditions to promote odontogenic differentiation of DPSCs and hMSCs in vitro and in vivo.…”

Fibrin-based scaffolds for dental pulp regeneration: from biology to nanotherapeutics

“…DP-MSC hypoxic preconditioning could also enhance angiogenesis by inducing VEGF expression (Aranha et al, 2010). Recently, exosomes isolated from human third molar DP cells, used as a delivery system for proteins, lipids, RNA and DNA, were proposed as another option to stimulate DP regeneration in a cell-free approach, since they stimulated the migration and proliferation of human bone marrow-derived MSCs in vitro in a fibrin hydrogel (Ivica et al, 2020).…”