Construction Of Dental Pulp Organoids Based On Prevascularized Scaffold Material

Based on the ACVM-0.25%HLC-I composite scaffold developed on the basis of previous studies, the CGF/GelMA composite material formed by modification of GelMA by CGF was combined to construct the prevascularized CGF/ GELMA-0.25% HLC-I composite scaffold. Vascular dental pulp organoids with spatial structure stability were constructed based on 3D cell oriented co-culture technology. GMSCs, VELCs and OLCs were seeded onto the CGF/GelMA-ACVM-0.25%HLC-I composite scaffold to construct vascularized dental pulp organoids. SEM, cyclic compression, and FTIR were used to analyze the ultrastructure and physicochemical properties. In vitro experiments, cell proliferation and migration were assessed via Calcein/PI staining, CCK-8, scratch and Transwell assays. The scaffold material's impact on cell viability was evaluated by a hemolysis assay, and the angiogenic potential was measured by tube formation assays. In vivo experiments, a bioreactor was used for semi-in situ pulp regeneration. The vascularized pulp organoid's morphology was identified by HE and Masson staining, while vascularization was assessed by immunofluorescence staining. The organoid's layered structure was traced by EdU Apollo 488/567, and blood flow signals were monitored by a small-animal ultrasound system. The biocompatibility and toxicity of scaffolds in vivo were tested for each important organ in bioreactor. The construction of vascularized dental pulp organoids is enabled by the 3D cell-laden modified CGF/GelMA-ACVM-0.25%HLC-I scaffold. Construction and regeneration of dental pulp organoids are greatly enabled by vascularization.