<p>Lutein, the main pigment in the macula, has been widely investigated for its role in retinal health and age-related macular degeneration. Although, its limited water solubility restricts effective ocular delivery. To address these limitations, the present study developed a lutein loaded nanostructured lipid carrier (Lu-NLCs) by applying a nano-template engineering strategy. The composition of Lu-NLCs was optimized using a two-factor, three-level central composite design (CCD) and further refined through a desirability function approach, wherein the effects of two independent variables, namely Compritol and olive oil concentrations, on zeta potential, particle size, entrapment efficiency (%EE) and polydispersity index (PDI), were evaluated. Further, the optimized Lu-NLCs_opt formulation was characterized using XRD, TEM, in-vitro drug release, FTIR analyses, ex-vivo permeation, HET-CAM, and histopathological studies. The thirteen formulated batches of Lu-NLCs demonstrated particle size values ranging from 320.7 to 784&#xa0;nm, %EE ranging from 45.78% to 75.02%, PDI ranging from 0.321 to 0.894, and zeta potential ranging from − 33.8 to -60.4 mV. The optimized Lu-NLCs_opt formulation demonstrated sustained in-vitro drug release, with 81.45% of the drug released over 12&#xa0;h. Ex-vivo permeation studies showed a 2.08-fold enhancement in drug uptake across the goat cornea in comparison to the plain drug solution. Furthermore, results from HET-CAM and histopathological evaluations confirmed that the Lu-NLCs_opt were safe, non-irritant, and biocompatible with ocular tissues. Overall, these findings confirm the successful development of Lu-NLCs using a CCD approach, demonstrating enhanced lutein permeation across excised corneas without any signs of ocular irritation, thereby highlighting their potential as well-tolerated carriers for ocular drug delivery.</p> Graphical Abstract <p></p>