The aim of this review was to consider laboratory experiments and clinical studies of calcium hydroxide-based root canal sealers. An extensive search of the endodontic literature was made to identify publications related to calcium hydroxide-based root canal sealers. The articles were assessed for the outcome of laboratory and clinical studies on their biological properties and physical characteristics. Comparative studies with other sealers were also considered. Several studies were evaluated covering different properties of calcium hydroxide-based sealers including physical properties, biocompatibility, leakage, adhesion, solubility, antibacterial properties, and periapical healing effect. Calcium hydroxide-based root canal sealers have a variety of physical and biological properties. Comparative studies reveal their mild cytotoxicity, but their antibacterial effects are variable. Further research is required to establish the tissue healing properties of calcium hydroxide in root canal sealers.