Biological properties of Ceraputty as a retrograde flling material: an in vitro study on hPDLSCs

Abstract

Objectives. To assess the cytocompatibility and bioactive potential of the new calcium silicate-based cement Ceraputty on human periodontal ligament stem cells (hPDLSCs) compared to Biodentine and Endosequence BC root repair material (ERRM). Materials and methods. hPDLSCs were isolated from extracted third molars from healthy donors. Standardized sample discs and 1:1, 1:2, and 1:4 eluates of the tested materials were prepared. The following assays were performed: surface element distribution via SEM–EDX, cell attachment and morphology via SEM, cell viability via a MTT assay, osteo/cemento/odontogenic marker expression via RT-qPCR, and cell calcifed nodule formation via Alizarin Red S staining. hPDLSCs cultured in unconditioned or osteogenic media were used as negative and positive control groups, respectively. Statistical analysis was performed using one-way ANOVA or two-way ANOVA and Tukey’s post hoc test. Statistical signifcance was established at p<0.05. Results. The highest Ca2+ peak was detected from Biodentine samples, followed by ERRM and Ceraputty. hPDLSC viability was signifcantly reduced in Ceraputty samples (p<0.001), while 1:2 and 1:4 Biodentine and ERRM samples similar results to that of the negative control (p>0.05). Biodentine and ERRM exhibited an upregulation of at least one cemento/odonto/osteogenic marker compared to the negative and positive control groups. Cells cultured with Biodentine produced a signifcantly higher calcifed nodule formation than ERRM and Ceraputty (p<0.001), which were also higher than the control groups (p<0.001). Conclusion. Ceraputty evidenced a reduced cytocompatibility towards hPDLSCs on its lowest dilutions compared to the other tested cements and the control group. Biodentine and ERRM promoted a signifcantly higher mineralization and osteo/cementogenic marker expression on hPDLSCs compared with Ceraputty. Further studies are necessary to verify the biological properties of this new material and its adequacy as a retrograde flling material. Clinical relevance This is the frst study to elucidate the adequate biological properties of Ceraputty for its use as a retrograde flling material.