Updated project metadata. The human cornea relies on its extracellular matrix (ECM) properties, with corneal stromal fibroblasts (CSFs) crucial for ECM remodeling. We studied the effects of nanofibrillar materials on CSFs using a decellularized human amniotic membrane (dHAM) and fibrillar collagen scaffold (FCS). FCS enhances focal adhesions (FAs), activating mechanotransduction signals through FAK autophosphorylation and YAP1 nuclear translocation. Nanofibrillar topography induces gene expression changes related to tissue remodeling and collagen production. Here, we performed shotgun proteomics analysis (DDA-PASEF in TimsToF Pro) to analyze the impact of FCS topography on CSFs protein expression.