Abstract
Abstract: :
Purpose: To measure proinflammatory, Th1-type and Th2-type cytokine and chemokine production in tears of myopic patients before and after LASIK surgery. Methods: Tear samples were obtained from 15 myopic patients (15 eyes, range –1D to -10D) before LASIK, and 1 and 24 hours after surgery. Concentrations of IL-1ß, -2, -4, -5, -6, -8, -10, -12, -13, IFNγ, TNFγ, Eotaxin, RANTES and MCP-1 were determined using multiplexed bead analysis. Samples were diluted 1:1 with buffer and incubated with a cocktail of LuminexTM beads, each coated with an individual monoclonal anti-cytokine/chemokine antibody, followed by biotinylated polyclonal antibodies and streptavidin-phycoerythrin. The fluorescence intensity of each bead population was measured using a Luminex100TM. The concentration of each molecule was determined from a standard curve of known concentrations of recombinant molecules. Results: The only chemokine found consistently at baseline was IL-8. IL-8, -1ß , -2, -6, -10, -12, –13, MCP-1 and RANTES mean tear levels were increased at 24 hours compared to baseline, however these differences were not statistically significant. Unexpectedly, eotaxin mean tear levels were significantly increased at 24 hours (74±36 pg/ml) compared to baseline (28±28 pg/ml; p<0.02). Interestingly, IL-12 mean tear levels were significantly increased 1 hour after surgery (14±6 pg/ml) compared to baseline (6±6 pg/ml; p<0.04). IL-4, IL-5 and IFNγ tear levels were below detection limits of the assay in 11 of 15 patients. Conclusions: Multiplexed bead analysis was shown to be a new and helpful method to measure multiple cytokine and chemokine production in tears. After LASIK, multiple cytokines may be released to modulate the corneal inflammatory and healing process.
Keywords: cytokines/chemokines • cornea: tears/tear film/dry eye • refractive surgery: LASIK