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Milk-derived beverages are considered to be beneficial to not only health but also to dental tissue. However, the exact nature of the effect of these beverages should be studied to formulate an exact diet plan for a child so that only good effects can be obtained from a dietary derivative., Aim and Objective: To evaluate and compare the surface roughness and microhardness of human enamel surface after exposure to different immersion regimes in milk-based beverages., Materials and Methods: A total of 90 samples were prepared from caries-free premolars extracted for orthodontic purposes. The samples were then randomly divided into three groups based on the immersion beverage used vis-a-vis chocolate milk, sweet lassi , and soy milk. The immersion cycles were done for 7 days. After the immersions were complete, the samples were dried and subjected to microhardness and surface roughness testing., Results: The surface roughness of the exposed surface of enamel was found to be in the following order: sweet lassi (verka) > control > soya milk chocolate (sofit) > chocolate milk (Amul Kool Koko). The microhardness of the exposed surface of enamel was found to be in the following order: chocolate milk (Amul Kool Koko) > soya milk chocolate (sofit) > sweet lassi (verka) > control., How to Cite This Article: Ghavri T, Pathak A, Bajwa NK. Comparative Evaluation of the Effect of Different Beverages on the Surface Roughness and Microhardness of Human Enamel Surface: An In Vitro Study. Int J Clin Pediatr Dent 2021;14(5):657-661., Competing Interests: Source of support: Nil Conflict of interest: None, (Copyright © 2021; Jaypee Brothers Medical Publishers (P) Ltd.)