Objective: Though e-cigarette is thought of as a healthy alternative way of smoking, the unknown nicotine concentration level and chemical compounds from the aerosol mist and flavourings are potentially harmful and may exert irritation or damage to the ocular surface. Thus, a cross-sectional study was conducted to identify the conjunctival and corneal endothelium profile among healthy e-cigarette smokers, and the relationship between smoking behaviour with the conjunctival and corneal endothelium profile. Material and Methods: Seventeen healthy e-cigarette smokers and 17 non-smokers were recruited for this study where their palpebral redness, bulbar redness, limbal redness, conjunctival roughness, central corneal thickness, endothelial cell density, and coefficient of variation of endothelium cell area were assessed using slit lamp biomicroscope and specular microscope. Results: There was a non-significant, higher mean rank observed in the total palpebral redness, total bulbar redness, total limbal redness, total conjunctival roughness, central corneal thickness, endothelial cell density, and coefficient of variation of the endothelium cell area in e-cigarette smokers compared to non-smokers (p-value>0.05). Only smoking frequency and total palpebral redness showed a statistically significant, positive, and fair correlation (r=0.349, p-value=0.043), while no other smoking behaviour showed a significant correlation (p-value<0.05). Conclusion: The use of e-cigarettes showed no significant impact on the conjunctival and corneal profile among the users, but further research with comprehensive evaluation is required in a larger sample size of more established e-cigarette smoking profiles.