Background: Tobacco smoke is toxic for cells and could be a damaging factor to skin. The purpose of this study was to compare the biophysical properties of skin in smokers and non-smokers. Materials and Methods: The study population consisted of 28 current smokers and 24 non-smokers. The hydration of the stratum corneum, trans epidermal water loss, pH, erythema, melanin content, sebum, friction and elasticity parameters (R0, R2, R5) of skin, epidermis and dermis thickness and echodensity were measured on middle forehead, right cheek and right inner arm of participants. Also volume, surface area and depth of right nasolabial folds were measured. The mean of these values in smokers were compared with nonsmokers by independent sample T- test. Results: Gross elasticity was significantly lower in smokers on forehead (p= 0.048). Thickness of epidermis was higher in smokers in all measured sites but the differences were not statistically significant. Thickness of dermis was higher in smokers in all measured sites too, but only the difference on cheek was statistically significant (p= 0.009). Density of epidermis was lower in smokers in all measured sites, but only the difference on forehead was statistically significant (p= 0.019). Density of dermis was lower in smokers in all measured sites, but only the difference on arm was statistically significant (p= 0.028). Volume and area of nasolabial folds were higher in smokers, but only the difference of area was statistically significant (p = 0.031). Conclusion: Tobacco smoking could affect the biophysical parameters of skin, especially thickness and density of dermis and epidermis and nasolabial folds.