Background. Dental caries is known as a multimicrobial disease. Caries are very prevalent in numerous countries, and the incidence is higher in underdeveloped countries than in developed countries. Dental caries is a major public health problem, and it is the most prevalent health problem across the world, affecting 2.4 billion people. Natural mouthwashes can be beneficial in the prevention of dental caries and oral infections without the side effects of synthetic mouthwashes. Aim. The aim of the present study was to investigate the antibacterial, antifungal, and cytotoxicity properties of sea salt from different areas of Iran. Methods and Materials. Sea salts from different areas (Urmia, Qom, and Jarquyeh) of Iran were collected. In order to define the elemental and mineralogical features of different salt samples, X-ray powder diffraction (XRD) was employed. Different concentrations (0.19–50 mg/mL) of sea salt were used in the antimicrobial and antibiofilm tests. The antimicrobial (MIC, MBC, MFC, and DAD tests) and antibiofilm (formation and degradation tests) effects were evaluated against L. acidophilus, S. aureus, E. coli, S. mitis, S. mutans, S. salivarius, and C. albicans. The cytotoxic effect of salts was evaluated on human gingival fibroblasts by the MTT test. Results. The range of MIC values in mg ml−1 was as follows: S. salivarius (50), S. mutans (50), S. mitis (50), L. acidophilus (12.5 to >50), C. albicans (50), E. coli (12.5 to 25), and S. aureus (12.5 to 25), while MBC values were, S. mutans (>50), S. salivarius (>50), S. mitis (>50), L. acidophilus (50 to >50), C. albicans (>50), E. coli (50), and S. aureus (50). MTT results showed that more than 50% of cell viability depends on decreasing the salt concentration (<1.56 mg/ml). Conclusion. Sea salts had significant antimicrobial effects on cariogenic bacteria and C. albicans. Therefore, sea salts can be a suitable candidate for mouthwash.