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From the Glaucoma Center, University of California San Diego, La Jolla, California.
PURPOSE. To investigate the effect of prostaglandins (PGs) on the permeability of human sclera in vitro.
METHODS. Twenty-three pairs of human eye bank eyes were studied. Circular pieces
of sclera were cultured in low-serum DMEM/F-12 media. Scleral hydration
was assessed by measuring wet and dry weight of scleral cultures
incubated with medium for 3 days and with Hanks’ buffered saline
solution (HBSS) for 4 hours. To assess scleral permeability,
organ-cultured scleral tissues were exposed to 100 to 500 nM
PGF2
, 17-phenyltrinor PGF2, or PhXA85
(the active form of latanoprost) for 1, 2, and 3 days. Scleral
permeability was measured using a two-chamber Ussing apparatus and
rhodamine-dextran polymers dissolved in HBSS (MW = 10,000, 40,000,
and 70,000). The movement of each rhodamine-dextran across the cultured
sclera was measured using a spectrofluorometer. To understand the
biological basis of the permeability change, the media were collected
from the treated cultures, and the concentration of MMP-1, 2, and 3 was
measured using an enzyme-linked immunosorbent assay.
RESULTS. There was no difference in scleral hydration among fresh sclera and
sclera incubated with medium for 3 days, with HBSS for 4 hours, or with
medium for 3 days followed by HBSS for 4 hours. Compared to tracer
movement across untreated scleral cultures (1.5 x
10-6 cm/sec for 10 kDa dextran, 0.7 x
10-6 cm/sec for 40 kDa dextran, and 0.4 x
10-6 cm/sec for 70 kDa dextran), exposure to
PGF2, 17-phenyltrinor PGF2, or PhXA85
each increased scleral permeability in a dose- and time-dependent
manner. Increases in permeability were greater with the10 kDa dextran
than with the 40 or 70 kDa dextran. The magnitude of these effects was
greatest with exposure to PhXA85 and similar with exposure to
PGF2 or 17-phenyltrinor-PGF2. MMP
expression also was significantly increased after PG exposure. These
increases were generally time and dose dependent and greater with MMP-2
and -3 than with MMP-1.
CONCLUSIONS. There is increased permeability of human sclera exposed to various PGs in organ culture. This increased permeability is accompanied by increased expression of MMPs.
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