Biomimetic Mineralized Tooth Constructs for Whole Tooth Bioengineering

Objectives: Globally, over 158 million people are suffering from tooth loss. Dental implants are currently the most common therapy intended to replace the function of lost teeth. However, complications may occur due to patient response to synthetic dental materials. The ability to engineer biomimetic replacement teeth would be a highly desirable alternative therapy. Here we describe a biomimetic three dimensional (3D) tooth model as a platform for the development of novel regenerative approaches for tooth replacement in humans. We have previously identified Gelatin Methacrylate (GelMA) hydrogel scaffold formulas that exhibit similar elastic moduli as natural dental tissues, and elicit optimized porcine dental epithelial (pDE) and porcine dental mesenchymal (pDM) cell morphology, metabolic activity and promote organized endothelial (HUVEC) structures in vitro. When implanted and grown subcutaneously, pDE/pDM/HUVEC constructs expressed dental cell differentiation markers, and exhibited tooth and bone specific mineralization and vascularization. Our objective here is to further optimize our biomimetic 3D tooth model by defining cell-seeding densities for optimized pDE and pDM proliferation and differentiation.
Methods: We will examine three cell-seeding densities (30,000 cells/µl, 60,000 cells/μl and 90,000 cells/ul). Replicate in vitro constructs will be prepared and grown for 1 and 14 days. Replicate samples will then be grown in vivo for 2 and 4 weeks. Histological, immunohistochemical, and radiographic analyses will be used to compare cellular organization, morphology, differentiation and biomineralization of both in vitro and in vivo constructs.
Results: We expect that the higher cell seeding density will improve dental cell interactions and dentin and enamel formation in biomimetic tooth constructs.
Conclusions: The results from this study will further establish dental cell encapsulated 3D GelMA hydrogel constructs as biomimetic 3D models for functional tooth replacement in humans.