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. 2024 Feb 22;24(1):262.
doi: 10.1186/s12903-024-04021-2.

Analysis of the cytotoxicity and bioactivity of CeraSeal, BioRoot™ and AH Plus® sealers in pre-osteoblast lineage cells

Luciano Aparecido de Almeida-Junior  1   2 , Giuliana de Campos Chaves Lamarque  2 , Henry Herrera  3 , Maya Fernanda Manfrin Arnez  2 , Francine Lorencetti-Silva  4 , Raquel Assed Bezerra Silva  2 , Léa Assed Bezerra Silva  2 , Francisco Wanderley Garcia Paula-Silva  5
Affiliations

Analysis of the cytotoxicity and bioactivity of CeraSeal, BioRoot™ and AH Plus® sealers in pre-osteoblast lineage cells

Luciano Aparecido de Almeida-Junior et al. BMC Oral Health. .

Abstract

Background: The objective of the present study was to evaluate in vitro the cytotoxicity and bioactivity of various endodontic sealers (CeraSeal, BioRoot™ and AH Plus®) in pre-osteoblast mouse cells (MC3T3 cells).

Methods: MC3T3 cells (ATCC CRL-2594) were plated in 1 × 104 cells/well in 96-well plates in contact with endodontic sealers at concentrations of 1:10 and 1:100. Cell viability was evaluated by MTT assay after 24 and 48 h. In addition, sealer bioactivity was measured by RT-PCR for mediator of inflammation (Tnf, Ptgs2) and mineralization (Runx2, Msx1, Ssp1 and Dmp1) after 24 h and by Alizarin Red S Assay of mineralization after 28 days. Data were analyzed using one-way ANOVA followed by the Tukey's post-test at a significance level of 5%.

Results: BioRoot™ presented 24-hour cytotoxicity (p < 0.05) at 1:10 concentration. In the period of 48 h, no endodontic cement was cytotoxic to the cells compared to the control (p > 0.05). TNF-α gene expression was induced by AH Plus® (p < 0.05), while Ptgs2 was induced by the CeraSeal and BioRoot™ (p < 0.05). The expression of Runx2 was stimulated by BioRoot™ and AH Plus® (p < 0.05). In contrast, the expression of Dmp-1 Dmp1 was higher for the CeraSeal and BioRoot™ (p < 0.05). Nonetheless, the sealers did not impact the formation of mineralization nodules (p > 0.05).

Conclusion: CeraSeal, BioRoot™ and AH Plus® sealers were not cytotoxic to MC3T3 cells within 48 h, but differentially induced the expression of genes related to inflammation and mineralization without impacting biomineralization by the cells.

Keywords: Biomineralization; Cytotoxicity; Inflammation; Osteoblasts; Root canal filling materials.

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Conflict of interest statement

The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
Percentage of cell viability by the MTT assay in a 24-hour period - control, DMSO, CeraSeal, Bio Root™ and AH Plus® groups at a concentration of 1:10 with αMEM (a). Percentage of cell viability by the MTT assay in a 24-hour period - control, DMSO, CeraSeal, Bio Root™ and AH Plus® groups at a concentration of 1:100 with αMEM (b). Percentage of cell viability by the MTT assay in a 48-hour period - control, DMSO, CeraSeal, Bio Root™ and AH Plus® groups at a concentration of 1:10 with αMEM (c). Percentage of cell viability by the MTT assay within 48 h - control, DMSO, CeraSeal, Bio Root™ and AH Plus® groups at a concentration of 1:100 with αMEM (d). Note: Different lowercase letters indicate that there is a statistical difference between the groups
Fig. 2
Fig. 2
mRNA expression for Tnf (a) and Ptgs2 (b) within 24 h after stimulation with CeraSeal, Bio Root™ and AH Plus® materials at a concentration of 1:100 or control (cells maintained with conventional culture medium). Note: Different symbols indicate that there is a statistical difference between the groups
Fig. 3
Fig. 3
mRNA expression for Runx2 (a), Dmp1 (b), Msx1 (c) and Spp1 (d) within 24 h after stimulation with CeraSeal, Bio Root™ and AH Plus® materials at a concentration of 1:100 or control (cells maintained with conventional culture medium). Note: Different symbols indicate that there is a statistical difference between the groups
Fig. 4
Fig. 4
Representative photomicrographs of Alizarin Red S Assay. Original 20x magnification (scale = 50 μm) (a). Formation of mineralization nodules at 28 days after stimulation with the CeraSeal, Bio Root™ and AH Plus® materials at a concentration of 1:100 or control (cells maintained with culture medium containing beta glycerophosphate and ascorbic acid) and DMSO (b). Note: Different symbols indicate that there is a statistical difference between the groups. Mineralization medium= beta-glycerophosphate and ascorbic acid
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