The Application of Pulp Tissue Derived-Exosomes in Pulp Regeneration: A Novel Cell-Homing Approach

Purpose: Exosomes derived from stem cells, as an alternative to stem cells themselves, have been employed for dental pulp regeneration. However, it is not known whether exosomes can recruit host cells to the regeneration process. In this study, we built a "cell homing" model to determine whether exosomes derived from dental pulp tissue (DPT-exos) can regenerate dental pulp by recruiting the stem cells from the apical dental papilla (SCAPs). Methods: Exosomes were isolated from the dental pulp tissue (DPT-exos) and dental pulp stem cells (DPC-exos) of swine. The effects of the exosomes on SCAPs were compared using CKK-8, Transwell, angiogenesis, and odontogenic induction assays. DPT-exos and DPC-exos were investigated in an in vivo "cell homing" model using swine teeth to compare their roles in pulp regeneration. To build the model, we placed SCAP-containing collagen gel at the root tip and filled the cavity of the treated dental matrix (TDM) with DPT-exos and DPC-exos-laden scaffolds, which would be expected to recruit SCAPs to the pulp cavity. The complex was then implanted subcutaneously into immunodeficient nude mice. After eight weeks, tissue samples were taken and analyzed histologically to determine whether the DPT-exos contributed to pulp regeneration through "cell homing". Results: Exosomes were successfully extracted from dental pulp tissue and confirmed to be exosomes. In vitro tests confirmed that DPT-exos performed better than DPC-exos in promoting the migration, proliferation, and differentiation of SCAPs. Furthermore, DPT-exos recruited SCAPs to regenerate dental pulp-like connective tissue in vivo containing collagen, odontoblasts, and enriched predentin-like tissue. Blood vessel growth was demonstrated by immunofluorescence. Conclusion: This study demonstrated the ability of DPT-exos to induce SCAPs to regenerate connective tissue similar to natural dental pulp. This technique has the potential for treating pulp deficiency caused by various pulp diseases.