In vitro evaluation of human dental pulp cells response to NeoMTA®2, ProRoot® MTA and TotalFill® BC RRM™ putty: biocompatibility and gene expression analysis

Aim To comparatively evaluate the effects of NeoMTA®2, ProRoot® MTA, and TotalFill® BC RRM™ Putty on human dental pulp cells (hDPCs) in terms of cell viability, attachment, odontogenic differentiation and biomineralization potential. Materials and Methods Dental pulp tissue was obtained from seven intact third molars extracted from six patients aged 18–23 years. Isolated hDPCs were cultured in direct contact with the tested materials. Cell viability was assessed using an MTT assay at 24, 48, and 72h. Gene expression of mineralization- and odontogenic differentiation–related markers was analyzed by real-time PCR on days 3, 5, and 7. Cell morphology and attachment were evaluated using scanning electron microscopy (SEM). Quantile regression models were used to investigate possible differences by material and time of measurement. Results All materials supported cell viability, with significantly higher MTT values at 72h compared with earlier time points. NeoMTA®2 induced significantly higher expression of alkaline phosphatase (ALP) and osteocalcin (OCN) at day 7 compared with ProRoot® MTA and TotalFill® BC RRM™ Putty. SEM analysis revealed abundant, well-spread cells firmly attached to all material surfaces. Conclusions NeoMTA®2 demonstrated cellular responses comparable to or greater than those of ProRoot® MTA and TotalFill® BC RRM™ Putty, favoring cell viability, attachment, and gene expression associated with mineralization and odontogenic differentiation. Clinical relevance: NeoMTA®2 shows promising biological properties that may support its use in vital pulp treatment; however, further laboratory and clinical studies are required.