Fig 1.
Preparation method of serum eye drops (SEDs) and solvent/detergent-treated (S/D)-SEDs.
For experimental details see the "Materials and Methods" section.
Fig 2.
SDS-PAGE under non-reducing conditions.
Molecular weight protein markers (left lane). Total protein content in each lane was normalized to 20 μg. Abbreviations: kDa: kilodaltons; SEDs: serum eye drops; S/D-SEDs: solvent/detergent-treated serum eye drops.
Fig 3.
Data from all groups after 3 weeks of treatment. Data are shown as the mean ± SD. * p<0.05 significantly from the control group (by One way ANOVA-Tukey’s multiple comparison. Duplicate experiments. Abbreviations: SEDs: serum eye drops; S/D-SEDs: solvent/detergent-treated serum eye drops.
Fig 4.
Slit-lamp photographs of rabbit eyes in each group after fluorescein staining.
The epithelial defect area appears as green patches. (a) Control: a non-treated eye with no fluorescent pigment, (b) dry-eye syndrome (DES; 0.1% BAC); (c) PBS-treated eyes; (d) serum eye drops (SEDs) and (e) solvent/detergent-treated (S/D)-SEDs. A dramatic improvement of the epithelial defect was observed in the two groups treated with SEDs and S/D-SEDs, in which treated eyes were similar to the control groups and with no staining.
Fig 5.
Representative pictures of HE staining of corneas from each of the five groups.
(a) Normal corneas (control): three to five epithelial layers can be observed. Damage to the epithelium was observed in (b) dry-eye syndrome (DES; 0.1% BAC) and (c) phosphate-buffered saline (PBS)-treated group: the layered structured was destroyed and sponge-like stroma were found; (d) serum eye drops (SEDs) and (e) solvent/detergent-treated (S/D)-SEDs were similar to control groups with multilayered epithelium in the apical part.
Fig 6.
TUNEL staining of corneal sections from each group.
(a) Control, (b) dry-eye syndrome (DES; 0.1% BAC), and (c) phosphate-buffered saline (PBS)-treated groups showed positive staining (blue) representing serious damage to the epithelium layer; (d) serum eye drop (SED) and (e) solvent/detergent-treated (SD)-SED groups were similar to the control groups with a healthy epithelium in the apical part.
Fig 7.
Variation in tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-1β, and IL-8 inflammatory marker expressions in corneas exposed to different treatments.
Data are expressed as the mean ± SD (error bars). * p<0.05 versus control group; # p<0.05 versus DES; % p<0.05 versus PBS; @ p<0.05 versus SED (One way ANOVA—Tukey’s multiple comparisons).