Various three-dimensional (3D) culture methods have been introduced to overcome the limitations of in vitro culture and mimic in vivo conditions. This study aimed to evaluate two microsphere-forming culture methods and a monolayer culture method. We evaluated cell morphology, viability, osteo-, adipo-, and chondrogenic differentiation potential of dental pulp stem cells (DPSCs) cultured in 3D culture plates: ultra-low attachment (ULA) and U-bottomed StemFit 3D (SF) plates, and a two-dimensional (2D) monolayer plate. RNA sequencing (RNA-seq) revealed differentially expressed gene (DEG) profiles of the DPSCs. In contrast to an increasing pattern in the 2D group, cell viability in 3D groups (ULA and SF) showed a decreasing pattern; however, high multilineage differentiation was observed in both the 3D groups. RNA-seq showed significantly overexpressed gene ontology categories including angiogenesis, cell migration, differentiation, and proliferation in the 3D groups. Hierarchical clustering analysis revealed a similar DEG regulation pattern between the 3D groups; however, a comparatively different DEG was observed between the 2D and 3D groups. Taken together, this study shows that DPSCs cultured in microsphere-forming plates present superior multilineage differentiation capacities and demonstrate higher DEG expression in regeneration-related gene categories compared to that in DPSCs cultured in a conventional monolayer plate.

This study aimed to evaluate the effect of a novel bioactive glass (BAG)-containing dentin adhesive on the permeability of demineralized dentin. Bioactive glass (85% SiO₂, 15% CaO) was fabricated using the sol-gel process, and two experimental dentin adhesives were prepared with 3 wt% silica (silica-containing dentin adhesive; SCA) or BAG (BAG-containing dentin adhesive; BCA). Micro-tensile bond strength (μTBS) test, fracture mode analysis, field-emission scanning electron microscopy (FE-SEM) analysis of adhesive and demineralized dentin, real-time dentinal fluid flow (DFF) rate measurement, and Raman confocal microscopy were performed to compare SCA and BCA. There was no difference in μTBS between the SCA and BCA (p > 0.05). Multiple precipitates were evident on the surface of the BCA, and partial occlusion of dentinal tubules was observed in FE-SEM of BCA-approximated dentin. The DFF rate was reduced by 50.10% after BCA approximation and increased by 6.54% after SCA approximation. Raman confocal spectroscopy revealed an increased intensity of the hydroxyapatite (HA) peak on the dentin surface after BCA application. The novel BAG-containing dentin adhesive showed the potential of both reducing dentin permeability and dentin remineralization.

Kim DW, et al. J Korean Acad Conserv Dent. 2005 Nov;30(6):493-504. http://dx.doi.org/10.5395/JKACD.2005.30.6.493

Kim KY, et al. Restor Dent Endod. 2013 May;38(2):85-89. http://dx.doi.org/10.5395/rde.2013.38.2.85

Bang JB, et al. Restor Dent Endod. 2013 May;38(2):79-84. http://dx.doi.org/10.5395/rde.2013.38.2.79