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To determine the in vitro transscleral permeability of antibiotics for posterior segment infection.Scleral sections from moist chamber-stored human globes were mounted in a 2-compartment perfusion chamber. Fluorescent-labeled vancomycin, polymyxin B, or penicillin G was added to the episcleral surface, while the choroidal side was slowly perfused with balanced salt solution (Alcon Laboratories, Inc., Ft. Worth, TX). The perfusate was collected and fluorescence was measured using a fluorometer. From the measurements, permeability (Ktrans) was calculated. Photomicrographs were taken with a fluorescent microscope to evaluate tissue absorption.The Ktrans values (cm/s, mean +/- standard error) were 6.66 +/- 1.46 x 10(-7) for vancomycin, 3.90 +/- 0.59 x 10(-7) for polymyxin B, and 1.89 +/- 0.21 x 10(-6) for penicillin G. The percent of antibiotic that diffused across the sclera from the donor chamber in 24 hours was 20.6 +/- 4.2 for vancomycin, 12.6 +/- 2.0% for polymyxin B, and 50.8 +/- 4.8% for penicillin G.This study shows that human sclera is more permeable to lower molecular weight, water-soluble penicillin G than to vancomycin or polymyxin B. The data suggests that a local, noninvasive, transscleral drug-delivery method may be reasonable for treating intraocular infections.