Fabrication of Vascularized DPSC Constructs for Efficient Pulp Regeneration

Abstract

Dental pulp regeneration is a promising approach to restore the vitality of necrotic teeth. We have previously reported the fabrication of scaffold-free cell constructs containing only dental pulp stem cells (DPSCs) and their ability to form pulp-like tissue in the pulpless tooth. However, the DPSC construct could not build pulp-like tissue with a full root length because it is difficult to induce blood vessels from a small root canal foramen. Therefore, we hypothesized that vascular structure could be preformed in the DPSC construct by employing endothelial differentiation capability of DPSCs, and vascularized constructs might facilitate dental pulp regeneration in the pulpless tooth. In this study, vascularized DPSC constructs were fabricated by inducing endothelial differentiation, and then we investigated the behavior of differentiated DPSCs, the internal structure of cell constructs, and their pulp regenerative ability in vivo. We observed that DPSCs positive for CD31 and von Willebrand factor were localized at the outer layer of constructs and formed a reticulated lumen structure. The cells constituting the outer layer of the construct expressed endothelial differentiation markers at higher levels than cells in the inner part. These results indicated that DPSCs in the outer layer differentiated into endothelial cells and formed vascular-like structures in the cell construct. Next, a vascularized DPSC construct was transplanted into the human pulpless tooth that was implanted into immunodeficient mice in the subcutaneous space. After 6 wk of implantation, the vascularized construct formed pulp-like tissues with higher density of human CD31-positive blood vessels when compared with specimens implanted with a DPSC construct without prevascularization. These results suggest that the vascular structure formed in the DPSC construct facilitated the blood supply and enhanced pulp regeneration. This study demonstrates that a vascularized DPSC construct is a prospective biomaterial as an implant for novel dental pulp regeneration.

Cannabidiol Rescues TNF-α-Inhibited Proliferation, Migration, and Osteogenic/Odontogenic Differentiation of Dental Pulp Stem Cells

“…Results of our study and reports from the literature indicate 2.5 µM Angiogenesis plays a crucial role in dentin/pulp regeneration and pulp revitalization as well as bone regeneration [44]. DPSCs express a plethora of angiogenic factors to induce angiogenesis directly or via endocrine effects [45][46][47]. Among these factors, VEGF is a key angiogenic factor overexpressed by DPSCs [48].…”

Role of Heparan Sulfate in Vasculogenesis of Dental Pulp Stem Cells

“…DPSCs were recently reported to be capable of differentiating into vascular endothelial lineages (Sasaki et al 2020; Katata et al 2021). The excellent vasculogenic capacity of DPSCs is of great interest when considering stem cell therapy involving dental pulp regeneration and/or vascular regenerative medicine (Li et al 2016; Granz and Gorji 2020; Chen et al 2022).…”

Endothelial cells during craniofacial development: Populating and patterning the head

“…Interestingly, dental pulp stem cells (DPSCs) can also be induced to differentiate into endothelial cells, by culture in gel moulds ( Sasaki et al, 2020 ). These DPSC endothelial cells can then support pulp-like tissue regeneration ( Katata et al, 2021 ). DPSCs are neural crest derived while endothelial cells are mesodermal, emphasising the plasticity of the neural crest in being able to form a wide variety of tissue types.…”