IADR Abstract Archives
Modelling vascularized dental pulp-like constructs in vitro
Objectives: The management of necrotic dental pulp is undergoing a paradigm shift towards dental pulp-derived stem cells (DPSC)-based strategies to regenerate pulp vitality, providing a promising alternative to traditional root canal therapy. However, a mature tooth with a narrow apical foramen restricts the exchange of nutrients and oxygen leading to a hypoxic microenvironment that limits cell growth and proliferation. Endothelial cell (EC)-laden pre-vascularized engineered pulp tissue constructs provide opportunities to accelerate angiogenesis and pulp regeneration. In this study, we investigated the fabrication of vascularized dental pulp-like constructs using a GelMA-fibrin hydrogel laden with DPSCs and ECs.
Methods: Dual-cure hydrogels were fabricated using varying concentrations of GelMA (6 to 10%) mixed with fibrinogen (6 mg/ml), followed by photocuring and enzymatic cross-linking. The ability of the hydrogels to support vasculature was evaluated by encapsulating the DPSC and microvascular ECs within the hydrogels followed by live/dead staining. To evaluate the potential for the formation of microvasculature, DPSC and ECs were seeded in different ratios (1:10, 10:1 and 1:1), cultured for eight days, and the tissue constructs were evaluated using confocal microscopy for vascular network parameters.
Results: The GelMA-fibrin hydrogel supported cellular viability and proliferation of ECs and DPSCs. Upon culture over 4-8 days, the DPSC-EC co-culture within these hydrogels resulted in the formation of an extensive network of microvasculature. The microvasculature demonstrated the presence of lumen, expression of vascular markers CD31 and vWF, supported by vimentin-expressing DPSCs and collagen-IV rich basement membrane.
Conclusions: The proof-of-concept study shows the potential to fabricate vascularized pulp-like constructs in vitro. Future studies on understanding the mechanics of vascularization under microenvironments representative of pulpal inflammation could shed more light on developing novel pulp regeneration strategies.
Methods: Dual-cure hydrogels were fabricated using varying concentrations of GelMA (6 to 10%) mixed with fibrinogen (6 mg/ml), followed by photocuring and enzymatic cross-linking. The ability of the hydrogels to support vasculature was evaluated by encapsulating the DPSC and microvascular ECs within the hydrogels followed by live/dead staining. To evaluate the potential for the formation of microvasculature, DPSC and ECs were seeded in different ratios (1:10, 10:1 and 1:1), cultured for eight days, and the tissue constructs were evaluated using confocal microscopy for vascular network parameters.
Results: The GelMA-fibrin hydrogel supported cellular viability and proliferation of ECs and DPSCs. Upon culture over 4-8 days, the DPSC-EC co-culture within these hydrogels resulted in the formation of an extensive network of microvasculature. The microvasculature demonstrated the presence of lumen, expression of vascular markers CD31 and vWF, supported by vimentin-expressing DPSCs and collagen-IV rich basement membrane.
Conclusions: The proof-of-concept study shows the potential to fabricate vascularized pulp-like constructs in vitro. Future studies on understanding the mechanics of vascularization under microenvironments representative of pulpal inflammation could shed more light on developing novel pulp regeneration strategies.