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. 2022 Oct 15;11(20):3238.
doi: 10.3390/cells11203238.

The Cytotoxicity and Genotoxicity of Bioactive Dental Materials

Marta Kunert  1 , Wioletta Rozpedek-Kaminska  2 , Grzegorz Galita  2 , Salvatore Sauro  3   4 , Rim Bourgi  5 , Louis Hardan  5 , Ireneusz Majsterek  2 , Monika Lukomska-Szymanska  1
Affiliations

The Cytotoxicity and Genotoxicity of Bioactive Dental Materials

Marta Kunert et al. Cells. .

Abstract

The promotion of biologically based treatment strategies in restorative dentistry is of paramount importance, as invasive treatments should be avoided to maintain the tooth's vitality. This study aimed to assess the biocompatibility of commercially available bioactive materials that can be used for dental pulp capping. The study was performed with a monocyte/macrophage peripheral blood SC cell line (ATCC CRL-9855) on the following six specific bioactive materials: ProRoot MTA (Dentsply Sirona), MTA Angelus (Angelus), Biodentine (Septodont), TheraCal LC (Bisco), ACTIVA BioACTIVE (Pulpdent) and Predicta Bioactive Bulk (Parkell). The cytotoxicity of the investigated agents was measured using a resazurin-based cell viability assay, while the genotoxicity was evaluated using an alkaline comet assay. Additionally, flow cytometry (FC) apoptosis detection was conducted with a FITC (fluorescein isothiocyanate) Annexin V Apoptosis Detection Kit I. FC cell-cycle arrest assessment was carried out with propidium iodide staining. The results of this study showed no significant cytotoxicity and genotoxicity (p > 0.05) in ProRoot MTA, MTA Angelus, Biodentine, ACTIVA BioACTIVE and Predicta Bioactive. Conversely, TheraCal LC presented a significant decrease (p < 0.001). In conclusion, due to excellent biocompatibility and low cytotoxicity, MTA, Biodentine, ACTIVA BioACTIVE and Predicta Bioactive may be suitable for pulp capping treatments. On the other hand, due to the high cytotoxicity of TheraCal LC, its use should be avoided in vital pulp therapies.

Keywords: cytotoxicity; dental materials; flow cytometry; genotoxicity; pulp capping; vital pulp therapy.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses or interpretation of data; in the writing of the manuscript or in the decision to publish the results.

Figures

Figure 1
Figure 1
Cytotoxicity of the investigated pulp capping materials. Tests performed using resazurin-based cell viability assay after 24 h (A) and 48 h incubation (B) of cells with the tested compounds. The positive control was represented by the cells incubated with 100% DMSO, while the negative control cells were cultured in a complete IMDM medium. Statistical significance on the graphs: *** p < 0.001 versus negative control.
Figure 2
Figure 2
Genotoxicity of the investigated materials. Analysis was performed using an alkaline version of the comet assay after 24 h (A) and 48 h (B) incubation of cells with the tested compounds. Cells suspended in 10% DMSO were used for the positive control. Cells suspended in 1 mL of complete culture medium were employed as the negative control. Statistical significance on the graphs: *** p < 0.001 versus negative control.
Figure 3
Figure 3
Flow cytometric FITC annexin V/propidium iodide (PI) double staining analysis of apoptosis after 24 h and 48 h incubation of cells with the tested compounds. Dot plot graphs indicate share of viable (FITC annexin V negative, PI negative), early apoptotic (FITC annexin V positive, PI negative) late apoptotic (FITC annexin V positive, PI positive) and necrotic (FITC annexin V negative, PI positive) cells (A). Percentage of cells in each group after 24 h are presented below (B) and 48 h incubation (C). ***—statistically significant difference (p < 0.001).
Figure 3
Figure 3
Flow cytometric FITC annexin V/propidium iodide (PI) double staining analysis of apoptosis after 24 h and 48 h incubation of cells with the tested compounds. Dot plot graphs indicate share of viable (FITC annexin V negative, PI negative), early apoptotic (FITC annexin V positive, PI negative) late apoptotic (FITC annexin V positive, PI positive) and necrotic (FITC annexin V negative, PI positive) cells (A). Percentage of cells in each group after 24 h are presented below (B) and 48 h incubation (C). ***—statistically significant difference (p < 0.001).
Figure 3
Figure 3
Flow cytometric FITC annexin V/propidium iodide (PI) double staining analysis of apoptosis after 24 h and 48 h incubation of cells with the tested compounds. Dot plot graphs indicate share of viable (FITC annexin V negative, PI negative), early apoptotic (FITC annexin V positive, PI negative) late apoptotic (FITC annexin V positive, PI positive) and necrotic (FITC annexin V negative, PI positive) cells (A). Percentage of cells in each group after 24 h are presented below (B) and 48 h incubation (C). ***—statistically significant difference (p < 0.001).
Figure 3
Figure 3
Flow cytometric FITC annexin V/propidium iodide (PI) double staining analysis of apoptosis after 24 h and 48 h incubation of cells with the tested compounds. Dot plot graphs indicate share of viable (FITC annexin V negative, PI negative), early apoptotic (FITC annexin V positive, PI negative) late apoptotic (FITC annexin V positive, PI positive) and necrotic (FITC annexin V negative, PI positive) cells (A). Percentage of cells in each group after 24 h are presented below (B) and 48 h incubation (C). ***—statistically significant difference (p < 0.001).
Figure 4
Figure 4
Flow cytometry (FC) analysis of cell cycle progression using propidium iodide (PI) staining after 24 and 48 h incubation (A) of cells with the tested compounds. Percentage of cells in each group after 24 h are presented below (B) and 48 h incubation (C). Cells treated with 1 µM nocodazole constituted a positive control. Cells cultured in the complete medium represented the negative control.
Figure 4
Figure 4
Flow cytometry (FC) analysis of cell cycle progression using propidium iodide (PI) staining after 24 and 48 h incubation (A) of cells with the tested compounds. Percentage of cells in each group after 24 h are presented below (B) and 48 h incubation (C). Cells treated with 1 µM nocodazole constituted a positive control. Cells cultured in the complete medium represented the negative control.
Figure 4
Figure 4
Flow cytometry (FC) analysis of cell cycle progression using propidium iodide (PI) staining after 24 and 48 h incubation (A) of cells with the tested compounds. Percentage of cells in each group after 24 h are presented below (B) and 48 h incubation (C). Cells treated with 1 µM nocodazole constituted a positive control. Cells cultured in the complete medium represented the negative control.
Figure 4
Figure 4
Flow cytometry (FC) analysis of cell cycle progression using propidium iodide (PI) staining after 24 and 48 h incubation (A) of cells with the tested compounds. Percentage of cells in each group after 24 h are presented below (B) and 48 h incubation (C). Cells treated with 1 µM nocodazole constituted a positive control. Cells cultured in the complete medium represented the negative control.
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References

    1. Kim Y., Lee D., Song D., Kim H.M., Kim S.Y. Biocompatibility and bioactivity of set direct pulp capping materials on human dental pulp stem cells. Materials. 2020;13:3925. doi: 10.3390/ma13183925. - DOI - PMC - PubMed
    1. de Mendonça Petta T., Pedroni A.C.F., Saavedra D.F., Faial K.D.C.F., Marques M.M., D’Almeida Couto R.S. The effect of three different pulp capping cements on mineralization of dental pulp stem cells. Dent. Mater. J. 2020;39:222–228. doi: 10.4012/dmj.2018-349. - DOI - PubMed
    1. Ghilotti J., Sanz J.L., López-García S., Guerrero-Gironés J., Pecci-Lloret M.P., Lozano A., Llena C., Rodríguez-Lozano F.J., Forner L., Spagnuolo G. Comparative surface morphology, chemical composition, and cytocompatibility of Bio-C repair, biodentine, and proroot MTA on hDPCs. Materials. 2020;13:2189. doi: 10.3390/ma13092189. - DOI - PMC - PubMed
    1. Akbiyik S.Y., Bakir E.P., Bakir S. Evaluation of the Bond Strength of Different Pulp Capping Materials to Dental Adhesive Systems: An In Vitro Study. J. Adv. Oral Res. 2021;12:286–295. doi: 10.1177/2320206821997983. - DOI
    1. Hardan L., Mancino D., Bourgi R., Alvarado-Orozco A., Rodríguez-Vilchis L.E., Flores-Ledesma A., Cuevas-Suárez C.E., Lukomska-Szymanska M., Eid A., Danhache M.-L., et al. Bond Strength of Adhesive Systems to Calcium Silicate-Based Materials: A Systematic Review and Meta-Analysis of In Vitro Studies. Gels. 2022;8:311. doi: 10.3390/gels8050311. - DOI - PMC - PubMed

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