Matrix stiffness regulates the physiology of the cells and plays an important role in maintaining its homeostasis. It has been reported to regulate cell division, proliferation, migration, extracellular uptake and various other physiological processes. The alteration in matrix stiffness has also been well reported in various disease pathologies. However, in ocular system, Keratoconus (KC) is an ideal model to study the effect of matrix stiffness on endocytosis since the progression of the disease is controlled by increasing the stromal elasticity. Our study using corneal epithelial and retinal pigment epithelial cell lines showed that ocular cells do respond to matrix stiffness by altering their morphology and endocytic uptake of FITC-Dextran 20 kDa. Further, by using KC epithelium as a clinical model, we hypothesize that change in stromal elasticity may also affect the endocytosis of KC epithelium. Our results clearly showed alteration in the expression of actin binding proteins such as Phosphorylated Cofilin, Profilin, Focal adhesion kinase, and Vinculin. Apart from cytoskeletal rearrangement proteins, we also observed endocytic proteins such as Clathrin, Caveolin1 and Rab 11 to be affected by matrix stiffness. Our study thus establishes connecting role between endocytosis and matrix stiffness which could be used to understand the pathophysiology of keratoconus that it is influenced by both mechanical and biochemical factors. (C) 2020 Elsevier Inc. All rights reserved.