Mineral trioxide aggregate affects cell viability and induces apoptosis of stem cells from human exfoliated deciduous teeth

Abstract

BackgroundMineral trioxide aggregate (MTA) is widely used for pulp-capping procedures in permanent teeth and as a gold standard material in endodontics. The aim of the study was to investigate the effect of MTA on cell viability and apoptosis when MTA is directly in contact with Stem Cells from Human Exfoliated Deciduous Teeth (SHEDs).MethodsMTA was mixed and coated in the bottom of a 24-well plate. SHEDs collected and cultured from normal exfoliated human deciduous teeth (passages 3–4) were seeded on square cover glasses. The glasses with seeded SHEDs were incubated in the plates with or without MTA coating. They were divided into four groups: MTA direct contact, direct control, MTA indirect contact, and indirect control. After 1, 2 and 3 days of culturing, cell morphology was observed and cell viability was assessed by the WST-1 cell cytotoxicity assay. TUNEL assay, immunofluorescent labeling and western blot analysis were used to study the effects of MTA on SHEDs apoptosis.ResultsMTA impaired cell viability of SHEDs in 1, 2 and 3 days, and the effect of direct contact was more severe. Cell apoptosis with positive Annexin V and TUNEL staining was noted when there was direct contact with MTA. Western blot analysis revealed that Bcl-2 and Bcl-xL decreased after SHEDs were in contact with MTA.ConclusionsThis study shows that direct contact with 1 week post-set MTA significantly decreases the viability of SHEDs and induced cell apoptosis. The results suggest that there is a possible cytotoxic effect of pulp tissue when there is direct contact with MTA. Different responses would be expected due to the strong alkaline characteristics of fresh mixed MTA.

Response of stem cells from human exfoliated deciduous teeth (SHED) to three bioinductive materials – An in vitro experimental study

“…The tissue tags were suspended in screw capped test tubes containing transport media (50:50 ratio of Dulbecco’s Modified Eagle’s Medium: F12 with HEPES buffer, and 1% antibiotic actinomycotic solution, Himedia laboratories, Mumbai, India). The samples were sent to the laboratory for isolation and culture of stem cells (Goorha and Reiter, 2017, Tsai et al, 2018). At laboratory, pulp tissues were minced in a petri-dish using sterile blade into small pieces and placed in prewarmed 37 °C washing medium (Delbecco’s phosphate buffered saline without calcium or magnesium and 1% antibiotic actinomycotic solution) to clean all the debris from the tissues.…”

“…Evidences from experimental studies have demonstrated effects of EMD, MTA and Biodentine on human dental pulp tissues either separately or in combination with other materials through various treatment modalities at variable success rates (Nowicka et al, 2013, Tsai et al, 2018). There are numerous research papers and reports available in the literature regarding biocompatibility of EMD, MTA and Biodentine, either compared alone or with each other in one or the other way (Araújo et al, 2018, Min et al, 2009).…”

Characterization of Living Dental Pulp Cells in Direct Contact with Mineral Trioxide Aggregate

“…Because of the high surface pH of the MTA, MTA causes serious cell damage to soft tissue [ 39 ]. In fact, a study using stem cells from exfoliated deciduous teeth revealed that MTA impaired cell viability and caused cell apoptosis when cells were in direct contact with MTA [ 25 ]. In the present study using DsRed-PPU7 cells, we demonstrated that cell numbers were markedly reduced after Day 3.…”

“…Studies related to these components were conducted on the biological functionality of MTA, such as biocompatibility [ 1 , 2 , 3 , 7 , 8 ], sealing ability [ 1 , 2 ], antibacterial qualities [ 9 , 10 ], tissue engineering [ 11 ] and the mechanism of inducing apatite formation [ 12 , 13 , 14 , 15 , 16 , 17 ]. In addition to research related to the characteristics of MTA using mesenchymal stem cells and human dental pulp cells, many studies were conducted to investigate the effect of MTA on cytotoxicity [ 18 , 19 ], cell proliferation [ 20 , 21 , 22 ], gene expression [ 21 , 23 , 24 ] and apoptosis [ 25 ]. However, there are few reports that examined the dynamics of living cells in direct contact with the MTA.…”

Differential Gene Expression Changes in Human Primary Dental Pulp Cells Treated with Biodentine and TheraCal LC Compared to MTA

“…Several studies have compared the biological effects of pulp capping materials on pulp cells [24,58]. However, these studies were limited to evaluating the expression of specific genes based on biological effects only.…”