Biocompatibility and osteogenic potential of novel tricalcium silicate-based materials in human dental pulp stem cells: Advancing vital pulp therapies

Objectives

To evaluate the cytocompatibility, bioactivity, and mineralization potential of new tricalcium silicate-based Material (Biodentine XP) on human dental pulp stem cells (hDPSCs) compared to other calcium silicate-based materials (MTA-Ang and Ther-PT).

Materials and methods

Standardized discs and 1:1, 1:2, and 1:4 eluates of Biodentine XP (BD-XP), MTA-Ang, and Ther-PT were prepared after setting. Human dental pulp stem cells (hDPSCs) were isolated from extracted third molars of healthy patients and cultured under standard conditions. The following assays were performed: cell attachment and morphology were assessed by scanning electron microscopy (SEM); metabolic activity and cell viability was evaluated by MTT and cell cycle analysis; cellular calcium ion content was analyzed by energy dispersive X-ray (EDX) and calcium release analysis; cell migration/proliferation by wound healing assays; cytoskeletal organization, analysis of cell apoptosis and necrosis, reactive oxygen species production, osteogenic marker expression were quantified by flow cytometry and RT-qPCR; and cell mineralization potential was determined through Alizarin Red S staining. Comparisons were made with hDPSCs cultured in unconditioned media (negative control) and osteogenic culture media (positive control). Statistical significance was set at p < 0.05.

Results

Biodentine XP (BD-XP) showed significantly positive results in cytocompatibility assays, including cell metabolic activity, viability, attachment, and morphology, compared to the negative control group, whereas MTA-Ang and Ther-PT showed moderate results. BD-XP exhibited a significant upregulation of osteogenic markers, including ALP and DSPP, compared to both the negative and positive control groups. In terms of mineralization potential, BD-XP-treated cells showed significantly higher calcified nodule formation compared to MTA-Ang, Ther-PT, the negative control, and the positive control groups.

Conclusion

BD-XP exhibits superior cytocompatibility and promotes osteo/odontogenic differentiation of human dental pulp stem cells (hDPSCs). BD-XP significantly enhances the upregulation of osteo/odontogenic markers such as ALP and DSPP, and promotes calcified nodule formation more effectively than MTA-Ang and Ther-PT. The high calcium ion content of BD-XP plays a key role in its ability to promote cellular adhesion, proliferation, and mineralization, making it a highly effective material for regenerative dental applications. These findings provide strong evidence to support the use of Biodentine XP in regenerative dental therapies, particularly in vital pulp treatments, where enhanced cellular adhesion and mineralization are critical for clinical success.