IADR Abstract Archives
Dentin Regeneration Using Bioceramic Scaffolds/Dental Stem Cells/Growth Factor Constructs
Objectives: This study aimed to investigate the potential of Mg-based bioceramic scaffolds combined with Dental Pulp Stem Cells-DPSCs and growth/morphogenetic factors (Dentin Matrix Protein-DMP-1 or Bone Morphogenetic Protein-BMP-2) to promote odontogenic differentiation and dentin-like tissue formation.
Methods: DPSCs cultures were established from third molars of young healthy donors. Human Treated Dentin Matrices-hTDMs were prepared from crowns of extracted third molars and demineralized in ultrasonic cleaner with EDTA. Sol-gel synthesized bioceramic scaffolds, consisting of 60 SiO2, 7.5 MgO, 30 CaO and 2.5 ZnO, all in wt %, were hosted in cavities inside hTDMs. DPSCs were spotted at 2×106 cells/scaffold/hTDMs complex and exposed to either 100 ng/ml DMP-1 or BMP-2 for 24h. Real time PCR was used to evaluate osteo/odontogenic markers' expression after 7 and 14 days (d), while Scanning Electron Microscopy-SEM and X-ray diffraction-XRD analysis were used for morphological and structural analysis of the regenerated tissues after 28 d of in culture.
Results: A significant up-regulation of several odontogenesis-related markers was observed for DPSCs grown inside the hTDMs/scaffold constructs (up to 26-fold for BMP-2, 28-fold for DSPP, 2.2-fold for ALP and 1.8-fold for BGLAP after 14 d). Exposure of DPSCs to either BMP-2 or DMP-1 for 24h was associated with a statistically significant increase of all odontogenesis genes' expression (p<0.01). XRD analysis showed the formation of an apatite layer on the surface of the scaffolds, while SEM-EDS analysis verified the formation of a thin layer with fibrous-like appearance with the presence of elements, including Ca, S and P.
Conclusions: Mg-based bioceramic scaffolds promoted odontogenic differentiation and dentin-like tissue formation. Moreover, DMP-1 and BMP-2 were proved beneficial to the odontoblastic differentiation of DPSCs inside the hTDMs/scaffold constructs, which makes them potential candidates for biofunctionalization on scaffold surfaces in future application for dental tissue engineering.
Methods: DPSCs cultures were established from third molars of young healthy donors. Human Treated Dentin Matrices-hTDMs were prepared from crowns of extracted third molars and demineralized in ultrasonic cleaner with EDTA. Sol-gel synthesized bioceramic scaffolds, consisting of 60 SiO2, 7.5 MgO, 30 CaO and 2.5 ZnO, all in wt %, were hosted in cavities inside hTDMs. DPSCs were spotted at 2×106 cells/scaffold/hTDMs complex and exposed to either 100 ng/ml DMP-1 or BMP-2 for 24h. Real time PCR was used to evaluate osteo/odontogenic markers' expression after 7 and 14 days (d), while Scanning Electron Microscopy-SEM and X-ray diffraction-XRD analysis were used for morphological and structural analysis of the regenerated tissues after 28 d of in culture.
Results: A significant up-regulation of several odontogenesis-related markers was observed for DPSCs grown inside the hTDMs/scaffold constructs (up to 26-fold for BMP-2, 28-fold for DSPP, 2.2-fold for ALP and 1.8-fold for BGLAP after 14 d). Exposure of DPSCs to either BMP-2 or DMP-1 for 24h was associated with a statistically significant increase of all odontogenesis genes' expression (p<0.01). XRD analysis showed the formation of an apatite layer on the surface of the scaffolds, while SEM-EDS analysis verified the formation of a thin layer with fibrous-like appearance with the presence of elements, including Ca, S and P.
Conclusions: Mg-based bioceramic scaffolds promoted odontogenic differentiation and dentin-like tissue formation. Moreover, DMP-1 and BMP-2 were proved beneficial to the odontoblastic differentiation of DPSCs inside the hTDMs/scaffold constructs, which makes them potential candidates for biofunctionalization on scaffold surfaces in future application for dental tissue engineering.