Title: 1718 - Progress in Creating a Biomimetic Dental Cell Sheet GelMA Tooth Bud
Nelson Monteiro (Presenter)
Weibo Zhang, Tufts University
Elizabeth Smith, Tufts University
Winnie Costa, Tufts University
Shantel Angstadt, Tufts University
Pamela Yelick, Tufts University
Objectives: Our objective was to characterize dental cell (DC) differentiation and mineralized dental tissue formation in a novel biomimetic three-dimensional (3D) tooth bud model consisting of dental epithelial (DE), dental mesenchymal (DM) and HUVEC cell sheets (DE-HUVEC, DM-HUVEC), combined with dental cell-HUVEC encapsulated gelatin methacrylate (GelMA) hydrogel scaffolds.
Methods: Porcine DM (0.114 x106 cell/cm2) or DE 0.228 x106 cell/cm2) cells were seeded onto UpCell thermo-responsive tissue culture plates. After 12 days, HUVECs (0.057x106/cm2) were seeded on top, and DE-HUVEC and DM-HUVEC cell sheets were harvested after 14 days. Stacked DE-HUVEC/DM-HUVEC cell sheets were then sandwiched between GelMA encapsulated DE-HUVEC and DM-HUVEC cell layers (ESM), and cultured in vitro for 7 days in osteogenic media and implanted and grown in vivo for 1, 3 and 6 weeks. Control constructs were also examined. In vivo mineralization was assessed and quantified using microCT (Bruker). Dental cell differentiation and neovascularization were assessed using immunohistological and immunohistochemical analyses.
Results: In vivo implanted 3D tooth bud constructs exhibited mineralized tissue formation of specified size and shape in ESM, DE-DM, M and MSM constructs, and mineralized tissue density increased over in vivo implantation time. Highest volumes of most dense mineralized tissues were observed in ESM constructs at 6 weeks. Histological evaluations of paraffin embedded and serial sectioned 3D biomimetic tooth bud constructs showed dental cell differentiation marker expression and neo-vasculature formation.
Conclusions: Dental cell sheet containing GelMA constructs exhibited the highest mineralized tissue formation as compared to other 3D tooth bud constructs. We propose our biomimetic 3D Gelma cell sheet tooth bud construct as a model to study dental cell differentiation and mineralized dental tissue formation leading to the formation of functional biomimetic replacement teeth.
This abstract is based on research that was funded entirely or partially by an outside source:
The submitter must disclose the names of the organizations with which any author have a relationship, the nature of the relationship, and the clinical or research area involved. The following is submitted: NONE