IADR Abstract Archives
Effects of Pulp Capping Materials on Macrophage Differentiation and Activity
Objectives: Objectives: Pulp capping materials have been shown to modulate pulp inflammation and regeneration. Depending on their stimulation, pulp fibroblasts have been shown to interact with macrophages modulating their differentiation into pro-inflammatory M1 or anti-inflammatory M2 phenotypes. Thus, upon carious lesion simulation, these fibroblasts have been shown to induce the M1 phenotype. Here, we studied the effect of subjecting pulp fibroblasts to tricalcium silicate-based materials on macrophage differentiation by investigating their cytokine secretion profile and phagocytic activity.
Methods: Methods:
Samples of Biodentine™ (Tricalcium silicate) and TheraCal LC® (Tricalcium silicate with resin) were placed in MEM medium (0.05cm2/mL) for 24h to obtain the materials’ extracts. Human pulp fibroblasts, obtained by the explant outgrowth method, were physically injured and incubated with Lipoteichoic Acid (LTA) to simulate a carious lesion, and incubated with the materials’ extracts to simulate direct pulp capping. After 24h, fibroblast supernatants were harvested and incubated with undifferentiated macrophages (M0) to investigate their differentiation into M1/M2 phenotypes. Secretion of pro-inflammatory TNF-α and anti-inflammatory Il-10 were analyzed by ELISA; phagocytic capacity of S.Mutans was assessed using Gentamycin protection assay. M0 macrophages chemically induced into M1 or M2 phenotypes were used as controls.
Results: Results: Both materials’ extracts significantly decreased macrophage pro-inflammatory TNF-α secretion as compared to injured/LTA-treated fibroblasts and M1 macrophages. Conversely, a significant increase in IL-10 secretion was observed with the materials’ extracts as compared to injured/LTA-treated fibroblasts and M1 macrophages but was 25% less than M2 secretion level. Use of materials’ extracts significantly decreased macrophage phagocytic activity as compared to injured/LTA-treated fibroblasts and M1 macrophages, but remained was comparable to that of M2.
Conclusions: Conclusions: The macrophage cytokine secretion profile and phagocytic capacity upon use of Biodentine™ and TheraCal LC® was comparable to anti-inflammatory M2 phenotype. These findings indicate that calcium silicate-based materials modulate macrophage differentiation, and may be of interest in targeting pulp regeneration.
Methods: Methods:
Samples of Biodentine™ (Tricalcium silicate) and TheraCal LC® (Tricalcium silicate with resin) were placed in MEM medium (0.05cm2/mL) for 24h to obtain the materials’ extracts. Human pulp fibroblasts, obtained by the explant outgrowth method, were physically injured and incubated with Lipoteichoic Acid (LTA) to simulate a carious lesion, and incubated with the materials’ extracts to simulate direct pulp capping. After 24h, fibroblast supernatants were harvested and incubated with undifferentiated macrophages (M0) to investigate their differentiation into M1/M2 phenotypes. Secretion of pro-inflammatory TNF-α and anti-inflammatory Il-10 were analyzed by ELISA; phagocytic capacity of S.Mutans was assessed using Gentamycin protection assay. M0 macrophages chemically induced into M1 or M2 phenotypes were used as controls.
Results: Results: Both materials’ extracts significantly decreased macrophage pro-inflammatory TNF-α secretion as compared to injured/LTA-treated fibroblasts and M1 macrophages. Conversely, a significant increase in IL-10 secretion was observed with the materials’ extracts as compared to injured/LTA-treated fibroblasts and M1 macrophages but was 25% less than M2 secretion level. Use of materials’ extracts significantly decreased macrophage phagocytic activity as compared to injured/LTA-treated fibroblasts and M1 macrophages, but remained was comparable to that of M2.
Conclusions: Conclusions: The macrophage cytokine secretion profile and phagocytic capacity upon use of Biodentine™ and TheraCal LC® was comparable to anti-inflammatory M2 phenotype. These findings indicate that calcium silicate-based materials modulate macrophage differentiation, and may be of interest in targeting pulp regeneration.