Multiplex Analysis of Cytokine Profiles in the Aqueous Humor of Glaucoma Filtration Surgery Models and Its Effect on Scarring

Purpose: To characterize the dynamic evolution of aqueous humor cytokine profiles after glaucoma filtration surgery (GFS) and to identify molecular signatures associated with bleb survival outcomes. Methods: GFS was performed on 26 rabbits' right eyes. Intraocular pressure (IOP), bleb morphology, and aqueous humor samples were collected before and after surgery. Thirty cytokines were quantified using RayBio Rabbit Cytokine Array Q30. Statistical analyses included Kaplan-Meier survival analysis, repeated-measures analysis of variance, independent samples t-tests or nonparametric tests, and Spearman correlation. Results: IOP decreased significantly in the early postoperative period and then gradually increased and stabilized. The mean bleb survival time was 15.19 ± 2.47 days. Among the 30 cytokines, 25 showed significant time-dependent changes, with peak times ranging from postoperative day 1 to day 21. After surgery, 12 cytokines were significantly higher in the early bleb failure group compared to the late failure group, with the greatest number and strongest negative correlations with bleb survival time observed on postoperative day 7, among which Contactin-1 (r = -0.6018), follistatin-related gene (FLRG; r = -0.5906), and thrombospondin-2 (TSP-2; r = -0.5847) exhibited the strongest correlations. Conclusions: GFS can induce time-dependent elevations in multiple cytokines in the aqueous humor, with high expression of factors (such as Contactin-1, FLRG, and TSP-2) on postoperative day 7 closely associated with earlier bleb failure. Translational Relevance: This study profiles post-GFS cytokine dynamics in rabbit aqueous humor, suggesting day 7 as a potential key window where Contactin-1, FLRG, and TSP-2 may act as scarring contributors, providing potential molecular targets and timing for future targeted interventions to improve the long-term success of filtration surgery.