Cytotoxicity and proliferation evaluation on fibroblast after combining calcium hydroxide and ellagic acid

J Adv Pharm Technol Res. 2021 Jan-Mar;12(1):27-31. doi: 10.4103/japtr.JAPTR_154_20. Epub 2021 Jan 9.

Abstract

Calcium hydroxide induces chronic inflammation and pulp tissue necrosis due to its high pH value. Ellagic acid is an anti-inflammatory and antioxidant flavonoid. Therefore, the effect of combining calcium hydroxide and ellagic acid must be researched to reduce cell damage due to the application of calcium hydroxide. The objective of the study was to determine the cytotoxicity and proliferation of fibroblasts after combining calcium hydroxide and ellagic acid with ratios of 99:1, 98:2, 97:3, 96:4, and 95:5. Calcium hydroxide and ellagic acid with different ratios were mixed with water and stirred. Rat gingival fibroblasts were prepared and incubated in two 96-well microplates. The control group and treatment groups (16 samples) were placed in the microplate and incubated for 1 and 3 days. An MTT assay test was performed, and the absorbance was observed using the ELISA reader with a wavelength of 540 nm. Following that, the cell viability was calculated. The results were tabulated and analyzed using a one-way ANOVA. For all treatment groups, the fibroblast cells showed a viability of higher than 50%. There was a significant increase (P < 0.05) in the fibroblast cell proliferation after combining calcium hydroxide and ellagic acid with ratios of 99:1 and 97:3. The combination of calcium hydroxide and ellagic acid is nontoxic. The treatment groups with ratios of 99:1 and 97:3 showed increased fibroblast cell proliferation.

Keywords: Calcium hydroxide; MTT assay; cytotoxicity; ellagic acid; proliferation of fibroblast cells.