Purpose <p>Pirfenidone possesses significant anti-inflammatory and anti-fibrotic properties, making it a promising therapeutic agent for ocular fibrotic conditions. However, its clinical application is limited due to a short half-life (&lt; 19&#xa0;min) in corneal tissue and poor ocular bioavailability. This study aimed to develop and characterize a nanostructured lipid carrier based in situ gel formulation for enhancing the ocular delivery and therapeutic efficacy of PFD.</p> Methods <p>NLCs were prepared via hot melt emulsification followed by probe sonication and incorporated into a gellan gum-based in situ gel system. A central composite design was used for formulation optimization. The optimized nanostructured lipid carrier formulation included Compritol^® 888 ATO (1.64%), Capmul^® MCM (0.32%), Poloxamer 188 (0.5%), and Tween 80 (0.5%). The formulation was characterized for particle size, polydispersity index, zeta potential, entrapment efficiency, in vitro drug release, ex vivo permeation, cytotoxicity, bio adhesion, and biocompatibility using Hen’s egg test using choriollantoic membrane assay. Stability studies were also conducted.</p> Results <p>The optimized Pirfenidone loaded nanostructured lipid carrier showed a particle size of 90.15 ± 10.2&#xa0;nm, PDI of 0.155 ± 0.014, zeta potential of -11.4 ± 1.2 mV, and entrapment efficiency of 90.43 ± 2.14%. In vitro release studies demonstrated sustained drug release (84.39 ± 3.41% over 12&#xa0;h). Ex vivo corneal and scleral permeation were 82.97 ± 3.01% and 77.01 ± 1.98%, respectively. Biocompatibility, cytotoxicity, and stability assessments confirmed the safety and robustness of the formulation.</p> Conclusion <p>The developed nanostructured lipid carrier based in situ gel offers a promising strategy for enhancing the ocular bioavailability and therapeutic potential of Pirfenidone, potentially overcoming the limitations of conventional topical administration.</p> Graphical Abstract <p></p>