Cytocompatibility, bioactive potential and antimicrobial activity of an experimental calcium silicate‐based endodontic sealer

Abstract

Aim To evaluate the cytocompatibility, bioactive potential, antimicrobial and antibiofilm activities of an experimental calcium silicate‐based endodontic sealer, in comparison with TotalFill BC Sealer and AH Plus. Methodology Cytocompatibility was assessed by methyltetrazolium (MTT) and neutral red (NR) assays, after exposure of the Saos‐2 cells to the sealer extracts (1 : 2, 1 : 4, 1 : 8, 1 : 16 and 1 : 32 dilutions) for 24 h. The sealers were manipulated and placed in 12‐well culture plates and exposed to ultraviolet light; then, 5 mL of DMEM without serum was added. Cell bioactivity was evaluated by alkaline phosphatase activity (ALP) and Alizarin red staining (ARS). Antimicrobial and antibiofilm activities were evaluated by direct contact test (DCT) on planktonic cells (DCTPC) and modified DCT on biofilm formed in bovine dentine blocks (MDCT). MTT, NR and ALP data were analysed by two‐way anova and Bonferroni tests; ARS data by anova and Tukey's tests; and the microbiological data by Kruskal–Wallis and Dunn tests (α = 0.05). Results The experimental sealer, TotalFill BC and AH Plus were not cytotoxic to Saos‐2, in comparison with the negative control (P > 0.05). Greater ALP was observed after 7 days of exposure of Saos‐2 to AH Plus and the experimental sealer (P < 0.05) when compared to the control. Significantly greater mineralized nodule production was observed for TotalFill BC and the experimental sealer (P < 0.05). In DCTPC, the experimental sealer and TotalFill BC were associated with a significantly greater reduction of E. faecalis (P < 0.05) and eliminated C. albicans. In MDCT, the experimental sealer and TotalFill BC had significantly greater antibiofilm efficacy (P < 0.05). Conclusions The experimental calcium silicate‐based sealer was cytocompatible, bioactive, antimicrobial against E. faecalis and C. albicans and effective against E. faecalis biofilms, with potential for use in root canal treatment.

In Vitro Effect of Putty Calcium Silicate Materials on Human Periodontal Ligament Stem Cells

“…Alizarin Red assay was assessed to evaluate mineralization capacity of the root repair cements; both cements produced more calcium deposits than the negative control, confirming the inductive effect of both root repair materials on the mineralization and differentiation of hPDLSCs. Previous reports have shown that putty calcium silicate materials induced mineralization [25,27]. Calcium silicates are the main components of both cements (Table 1) which justifies the result obtained.…”

“…TFB and BCR showed similar results in terms of cell viability as compared with a control group. In line with this work, previous reports have described similar results, also using the MTT test with TotalFill BC sealer, which has an almost identical composition, except it changes consistency [25,26]. Benetti et al [27] also reported that BCR is a biocompatible material that induces mineralization in vivo.…”

“…We used the MTT test due to its ease of performing and its accuracy [25]. In addition, the methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay is commonly accepted for in vitro cytotoxic studies.…”

Physical Properties, Antimicrobial Activity and In Vivo Tissue Response to Apexit Plus

“…Our findings showed that Apexit Plus had an effective antibacterial action against Enterococcus faecalis, a facultative anaerobic Gram-positive coccus responsible for root canal treatment failures [34]. Here, the findings were obtained using the direct contact test for 60 minutes, a widely used methodology performed to assess the antimicrobial activity of materials [35,36]. The antimicrobial properties of calcium hydroxide-based sealers has been related to hydroxyl ion release and, consequently, to an increased alkaline pH in the environment promoted by fresh sealers [4].…”

MTT versus other cell viability assays to evaluate the biocompatibility of root canal filling materials: a systematic review

“…Since Mosmann (1983) proposed a fast tetrazolium salt-based colorimetric test (MTT) with applicability for the evaluation of cell viability/cell proliferation, this and other tetrazolium salt-based tests such as XTT (Scudiero et al 1988), MTS (Barltrop et al 1991) and WST-1 (Tan & Berridge 2000) have been widely used in the screening of cytotoxic effects of diverse substances (Vbrova et al 2016, Tao et al 2016 and materials, including those intended for root canal filling (Osorio et al 1998, Zordan-Bronzel et al 2019. These tests are based on the ability of metabolically active cells to produce reduced pyridine nucleotides, which act as substrates for cellular oxidoreductases, and are responsible for the process of reduction of tetrazolium salts into formazan salts (Tan & Berridge 2000).…”