BACKGROUNDElevated intraocular pressure (IOP) is primarily due to increased aqueous outflow resistance, but how aqueous outflow resistance is generated and regulated are still not fully understood. The aim of this study is to determine whether changes in outflow facility, outflow pattern, and morphology following acute IOP elevation were reversible when the IOP was returned to a normal level in bovine eyes using a two-color tracer technique to label outflow patterns within the same eye.

METHODSTwelve fresh enucleated bovine eyes were perfused with Dulbecco's phosphate buffer saline (PBS) containing 5.5 mmol/L glucose (DBG) at 30 mmHg first to establish the baseline outflow facility followed by a fixed volume of red fluorescent microspheres (0.5 µm, 0.002% v/v). After the red tracer being replaced with DBG in the anterior chamber, perfusion was continued at 7 mmHg with the same volume of green tracer, followed by a fixative. In two control groups, the eyes were constantly perfused at either 30 mmHg (n = 6) or 7 mmHg (n = 6) using the same methods. The outflow facility (C, µl × min × (-1)mmHg(-1)), was continuously recorded. Confocal images were taken along the inner wall (IW) of the aqueous plexus (AP) in frontal sections. The percent of the effective filtration length (PEFL, PEFL = IW length exhibiting tracer labeling/total length of IW) was measured. Sections with AP were processed and examined by light microscopy. The total length of IW and the length exhibiting separation (SL) in the juxtacanalicular connective tissue (JCT) were measured. A minimum of eight collector channel (CC) ostia per eye were analyzed for herniations.

RESULTSIn the experimental (30 - 7 mmHg) group, the outflow facility was significantly higher at 7 mmHg ((4.81 - 1.33) µl × min × (-1)mmHg(-1)) than that at 30 mmHg ((0.99 ± 0.15) µl × min × (-1)mmHg(-1), P = 0.002), corresponding to a significant increase in the PEFL (P = 0.0003). The percent of CC ostia exhibiting herniations in the experimental group ((67.40 ± 8.90) µl × min × (-1)mmHg(-1)) decreased significantly compared to that in the control at 30 mmHg ((94.44 ± 3.33) µl × min × (-1)mmHg(-1), P = 0.03), but higher than that in the control at 7 mmHg ((29.43 ± 4.60) µl × min × (-1)mmHg(-1), P = 0.01). Washout-associated separation between the IW and JCT was found by light microscopy and percent separation length (PSL, PSL = SL/total length of IW) was decreased in the control at 30 mmHg compared to that in the experimental group and control at 7 mmHg.

CONCLUSIONSThe pressure-induced morphological and hydrodynamic changes were reversible. Changes (collapse of AP, separation between the JCT and IW, and herniation into CC ostia) influence the effective filtration area that regulates outflow facility.