Fig. 3. Mechanisms of enhanced ophthalmic delivery of FCS/IgG nanocomplexes.

(A) Immunofluorescence imaging indicating the distribution of tight junction–related proteins, including occludin and E-cadherin, after different treatments (scale bars, 10 μm). (B to D) Western blotting images showing the expression of occludin, E-cadherin, MLC, and p-MLC and statistical analysis of MLCs and p-MLC in HCEC monolayers after incubation with different nanocomplexes. ns, not significant. (E) TEER of HCECs after different treatments for 1 hour. (F) Schematic showing the in vitro simulated corneal epithelial barrier consisting of an HCEC monolayer in the upper chamber and PDL1-expressing B16F10 cells in the lower chamber of a Transwell. (G) Intensity of fluorescence-labeled aPDL1 that combined with the remaining unblocked PDL1 antigen expressed in B16F10 cells. (H) TEM images of rabbit corneal epithelial tissue after treatment with PBS, SDS, FCS, or CS (scale bars, 5 μm) and magnified TEM images (scale bar, 500 nm). (I) Confocal images of mouse eyeballs 6 hours after FCS-FITC/IgG-Cy5.5, CS-FITC/IgG-Cy5.5, or free IgG-Cy5.5 eyedrops were applied. The cornea, iris, and retina were partially magnified (scale bars, 500 μm). Data were represented as means ± SD. P values in (D) and (G) were calculated by using t test (*P < 0.05).