Ideal storing solutions of teeth during in vitro studies must not affect the physical and chemical properties of dental tissues. Demineralized water (DW) and artificial saliva (AS) have been used in many in vitro studies. AS is used to resemble oral cavity environment, where as DW is used to insure no reaction between tested material and components of the storing solution. Many studies explained DW as minimal invasive solution on dental tissues used as control to compare it with tested groups. Objective: This study tried to provide accurate information about effect of DW and AS on roughness, morphology and microhardness of enamel surface. Methods: Bovine enamel samples (Ø=5mm) were embedded in acrylic resin blocks, finished with sand papers (1200, 2400, 4000) and polishing paste (0.1µm), and examined with light microscope (50x) for any defect on the surface after finishing. Two groups (n=12 /Group) were stored in DW and AS, respectively. Qualitative and quantitative changes of enamel surface concerning roughness (RA, RZDIN, RT_RY) were observed using laser profilometry before storing, after 1, 5, 11, 18 and 26 days of storage. Observation with scanning electron microscope and enamel microhardness were achieved before and after 26 days. Data were analyzed statistically using "Linear Mixed Effects Model for Repeated Measures Data". Results: No significant changes were observed over time during storage in AS. However, mean roughness values increased during storage in DW significantly (p<0.0001). The RA-values after 0, 1, 5, 11, 18, and 26 days, respectively, were 0.038 ±0.008, 0.057 ±0.01, 0.073 ±0.005, 0.12 ±0.02, 0.13 ±0.018, and 0.14 ±0.02 µm. Additionally enamel micro hardness was reduced significantly during the 26 days-storage in DW (254.8 ±21.9, 194.5 ±35.9 KHN, p=0.0061). Conclusion: Results of studies using DW as storing solution and examining influences on surface properties of enamel will have to be reevaluated.

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