Aim and objective: Cytotoxicity of polyvinyl siloxane (PVS), polyether (PE), and polyvinyl ether silicone (PVES) on NIH/3T3 cells. Materials and methods: This in vitro study used elastomeric impression materials which were divided into three groups, group I, II, and III with PVES (EXA\'lence light body), PVS (Flexceed light body), and PE impression material (Impregum), respectively. A total of 10 specimens were prepared. Dulbecco's modified Eagle's medium was used for growing mouse cell line NIH/3T3. Cytotoxicity level of all elastomers was measured with the test 3-(4,5-dimethylthiazol-2-yl)-2-5-diphenyltetrazolium bromide assay at regular intervals. Results: There was a decline in the survival rate with PVES as found on day 1, PVS and PE showed on 3rd and 7th day. Kruskal–Wallis test showed a significant difference in all groups at various days (p < 0.05). Conclusion: Authors found that PVES showed early cytotoxic signs as compared to PVS and PE. Cell viability for PVS was highest as compared to PVES and PE impression materials. Clinical implication: Cell viability for PVS was highest as compared to PVES and PE impression materials. This information is useful in the selection of impression materials.