IADR Abstract Archives
Analysis of Biocompatibility of Bioceramic Cements on CD4+Tcells
Objectives: The aim of this study was to evaluate the effect of Bioceramic cements on proliferation, viability and cytokine secretion of polyclonal activated CD4+Tcells.
Methods: Bioceramic cements are recently-developed endodontic cements that exhibit several bioactive characteristics and improved biocompatibility in comparison with widely used resin-based cements. However, up to date, there are no studies aimed atevaluatingtheir biocompatibility and bioactivity on human CD4+Tcells. Bioceramic cements Total Fill Putty, Biodentine, Total Fill Sealer, Bioroot-RCS and resin-based cement Top Seal were prepared according to manufacturer of instructions under sterile conditions. After preparation, cement extracts were obtained after incubating 3.5x1.5mm disc cement in 1ml of RPMI for 7 days at 37°C. Next, cement extracts were span at 3.500rpm, filtered with 0.45um and stored at -80°C for further use. Human CD4+ T cells were isolated from oral tissue biopsies using cell sorting and expanded with aCD3CD28 beads. Then, CD4+ T cells were cultured with or without each cement extract (25%v/v) for 5 days. Proliferation, viability and cytokine secretion were assessed using Cell Trace Violet, Live-dead dye and Cytokine Bead Array, respectively. Samples were acquired on a FortessaX20 and analyzed with FlowJo.
Results: Our data showed that in terms of viability, the extract of Total Fill Putty did not affect the viability of CD4+Tcells, whereas Biodentine, Total Fill Sealer and Bioroot-RCS reduced percentages and numbers of cells. Top Seal was the most cytotoxic cement, inducing a drastic reduction of live cells. On the other hand, proliferation rates were not affected in the remaining live CD4+Tcells cultured with cement extracts. Finally, cytokine secretion was compared between CD4+Tcells alone or in the presence of Total Fill Putty and the data revealed that the production of cytokines was affected by the presence of the former cement extract.
Conclusions: Our data demonstrated differences in biocompatibility between Bioceramic cements on CD4+Tcells cells.
Methods: Bioceramic cements are recently-developed endodontic cements that exhibit several bioactive characteristics and improved biocompatibility in comparison with widely used resin-based cements. However, up to date, there are no studies aimed atevaluatingtheir biocompatibility and bioactivity on human CD4+Tcells. Bioceramic cements Total Fill Putty, Biodentine, Total Fill Sealer, Bioroot-RCS and resin-based cement Top Seal were prepared according to manufacturer of instructions under sterile conditions. After preparation, cement extracts were obtained after incubating 3.5x1.5mm disc cement in 1ml of RPMI for 7 days at 37°C. Next, cement extracts were span at 3.500rpm, filtered with 0.45um and stored at -80°C for further use. Human CD4+ T cells were isolated from oral tissue biopsies using cell sorting and expanded with aCD3CD28 beads. Then, CD4+ T cells were cultured with or without each cement extract (25%v/v) for 5 days. Proliferation, viability and cytokine secretion were assessed using Cell Trace Violet, Live-dead dye and Cytokine Bead Array, respectively. Samples were acquired on a FortessaX20 and analyzed with FlowJo.
Results: Our data showed that in terms of viability, the extract of Total Fill Putty did not affect the viability of CD4+Tcells, whereas Biodentine, Total Fill Sealer and Bioroot-RCS reduced percentages and numbers of cells. Top Seal was the most cytotoxic cement, inducing a drastic reduction of live cells. On the other hand, proliferation rates were not affected in the remaining live CD4+Tcells cultured with cement extracts. Finally, cytokine secretion was compared between CD4+Tcells alone or in the presence of Total Fill Putty and the data revealed that the production of cytokines was affected by the presence of the former cement extract.
Conclusions: Our data demonstrated differences in biocompatibility between Bioceramic cements on CD4+Tcells cells.