IADR Abstract Archives
3D-Printed Bone Scaffold in a Tissue Engineered Human Osteo-Mucosal Model
Objectives: To develop and characterise three-dimensional tissue-engineered models of bone and oral mucosa using 3D printed (3DP) bone scaffolding techniques in comparison with the conventional scaffolding technique.
Methods: Two different tricalcium phosphate (TCP) scaffolds (2 mm × 10 mm) (N=15) were fabricated by (A) 3DP technique and (B) foaming technique. Human osteoblasts (HOBs) were harvested from the alveolar bone biopsies, expanded in vitro, seeded into both scaffold types, and the bone constructs were cultured in an osteogenic culture medium using spinner bioreactors for 20 days.
Full-thickness oral mucosa models (OMMs) were simultaneously fabricated through three stages: (1) Preparation and culture of collagen gel-embedded normal human oral fibroblasts; (2) addition of normal human oral keratinocytes onto the engineered connective tissue layer, and (3) air/liquid interface culture in a suitable epithelial differentiation medium. Following the initial culture, OMMs was adhered onto the bone constructs using a tissue adhesive sealant to produce composite osteo-mucosal models. The final reconstructs were kept in culture for additional 5 days after which PrestoBlue (PB) vitality assay, histological examination, enzyme-linked immunosorbent assay (ELISA), and q-PCR analysis were carried out to assess both hard and soft tissue differentiation.
Results: PB assay indicated the high cellular vitality in both 3DP and conventional models. The histological sections showed the epithelial, connective tissue, and bony layers which were comparable to the native tissue structure. q-PCR analysis showed the gene expression of bone-specific markers including; osteocalcin, osteonectin, osteopontin, alkaline phosphatase and collagen-1. In addition, the epithelial markers including cytokeratin 10, cytokeratin 13, and Ki-67 were expressed. ELISA detected the secretion of bone proteins; collagen-1, osteonectin, and osteocalcin in both tissue engineered models.
Conclusions: Tissue-engineered composite models developed in this study resembled the natural alveolar bone and oral mucosa and have the potential to be used in vivo and as a relevant model for various in vitro applications including biological evaluation of implanted biomaterials and oral disease modelling.
Methods: Two different tricalcium phosphate (TCP) scaffolds (2 mm × 10 mm) (N=15) were fabricated by (A) 3DP technique and (B) foaming technique. Human osteoblasts (HOBs) were harvested from the alveolar bone biopsies, expanded in vitro, seeded into both scaffold types, and the bone constructs were cultured in an osteogenic culture medium using spinner bioreactors for 20 days.
Full-thickness oral mucosa models (OMMs) were simultaneously fabricated through three stages: (1) Preparation and culture of collagen gel-embedded normal human oral fibroblasts; (2) addition of normal human oral keratinocytes onto the engineered connective tissue layer, and (3) air/liquid interface culture in a suitable epithelial differentiation medium. Following the initial culture, OMMs was adhered onto the bone constructs using a tissue adhesive sealant to produce composite osteo-mucosal models. The final reconstructs were kept in culture for additional 5 days after which PrestoBlue (PB) vitality assay, histological examination, enzyme-linked immunosorbent assay (ELISA), and q-PCR analysis were carried out to assess both hard and soft tissue differentiation.
Results: PB assay indicated the high cellular vitality in both 3DP and conventional models. The histological sections showed the epithelial, connective tissue, and bony layers which were comparable to the native tissue structure. q-PCR analysis showed the gene expression of bone-specific markers including; osteocalcin, osteonectin, osteopontin, alkaline phosphatase and collagen-1. In addition, the epithelial markers including cytokeratin 10, cytokeratin 13, and Ki-67 were expressed. ELISA detected the secretion of bone proteins; collagen-1, osteonectin, and osteocalcin in both tissue engineered models.
Conclusions: Tissue-engineered composite models developed in this study resembled the natural alveolar bone and oral mucosa and have the potential to be used in vivo and as a relevant model for various in vitro applications including biological evaluation of implanted biomaterials and oral disease modelling.