PMC

AlamarBlue assay to determine the viability of cells treated with eyesalve. (A) HaCaT cells and (B) THP-1 cells were treated with six batches of undiluted and 1/10 dilution of the eyesalve (B1–B6) for 24 h; the controls were cells only (untreated), Neosporin (Neo), a safe antibiotic for wound infections and Optrex chloramphenicol (chl) treated cells. The experiment was conducted twice on fresh cultures of cells, with n = 3 or 4 replicates per treatment. Squares and circles denote the two biological repeats, and the colour of eyesalve symbols denotes the batch tested. Note that all replicate HaCaT cultures treated with undiluted chloramphenicol or eyesalve showed no reduction of alamarBlue, thus not all symbols are visible. For HaCaT cells, Neosporin did not affect alamarBlue reduction relative to untreated cells, but alamarBlue reduction was lowered when cells were treated with diluted eyedrops (ANOVA on control treatments only, eliminating experiment: treatment F_2,15 = 17.5, p < 0.001; Dunnett’s test vs. cells only: Neosporin p = 0.445, diluted eyedrops p = 0.001). All batches of diluted eyesalve caused a greater drop in alamarBlue reduction than diluted eyedrops (ANOVA on diluted treatments only, eliminating experiment: treatment F_6,20 = 24.8, p < 0.001; Dunnett’s test vs. diluted eyedrops: all p ≤ 0.012). For THP-1 cells, both Neosporin and chloramphenicol eyedrops reduced alamarBlue reduction relative to untreated cells, but diluted eyedrops did not significantly affect alamarBlue reduction (ANOVA on control treatments only, eliminating experiment: treatment F_3,20 = 53.6, p < 0.001; Dunnett’s test vs. cells only: Neosporin p = 0.016, eyedrops p < 0.001, diluted eyedrops p = 0.517). When batches of undiluted eyesalve were compared with undiluted eyedrops, there was heterogeneity between the six batches (ANOVA on undiluted treatments only, eliminating experiment: treatment F_6,20 = 6.68, p < 0.001; Dunnett’s test vs. eyedrops: B1, B2, B4 p ≤ 0.011, B3, B5, B6 p ≥ 0.071). All batches of diluted eyesalve caused a greater reduction in viability than diluted eyedrops (ANOVA on diluted treatments only, eliminating experiment: treatment F_6,20 = 46.4, p < 0.001; Dunnett’s test vs. diluted eyedrops: all p < 0.001).

Group definitions. Group 1, autologous serum eyedrops (ASEs) for 2 months without discontinuing glaucoma eyedrops (n = 18); Group 2, ASEs for 2 months and discontinuing glaucoma eyedrops for the first month (n = 22); Group 3, preservative-free artificial tears for 2 months and discontinuing glaucoma eyedrops for the first month (n = 15).

Percentage of patients with increased intraocular pressure (IOP) after treatment in each group. Group 1, autologous serum eyedrops (ASEs) for 2 months without discontinuing glaucoma eyedrops (n = 18); Group 2, ASEs for 2 months and discontinuing glaucoma eyedrops for the first month (n = 22); Group 3, non-preservative artificial tears for 2 months and discontinuing glaucoma eyedrops for the first month (n = 15).

(A) Bilateral eczematous periorbital swelling after instillation of dorzolamide eyedrops. (B) Improved periorbital swelling after cessation of dorzolamide eyedrops.