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The changes induced in the microstructure and chemical composition of root dentin by irrigants are an important aspect and could have adverse implications such as decreased microhardness, flexural strength, modulus of elasticity, and surface energy of root dentin. These changes have a deleterious effect on the sealing ability and adhesion of resin-based cement and root canal sealers to dentin.Therefore, we conducted a study to evaluate and compare the calcium loss from root dentin and microhardness of root dentin using atomic absorption spectrophotometry and the Vickers hardness test respectively, following different irrigation regimens.