Morphological observations of rat corneal endothelial cells after exposure to ozonated solution.
OBJECTIVE
To determine whether exposure to ozonated solution alters the morphology of corneal endothelial cells in rats and to examine the protective effect of ascorbic acid.


METHODS
The anterior chambers of rat eyes were filled with 4 ppm of ozonated solution. Some were left in that state, while others were flushed out either 10, 30, or 60 s after exposure to a balanced salt solution (BSS), or to BSS containing 0.001 M ascorbic acid. Corneal endothelial cells were assessed by scanning and electron microscopy either 1 h or 1 week after treatment, and the expressions of aquaporin (AQ)-1 and zonula occludens (ZO)-1 were determined by immunohistochemistry.


RESULTS
When exposure time was longer than 10 s, damaged cell membranes and abnormal organelles were observed 1 h after treatment. The longer the exposure time, the more severe the observed alterations; however, the eyes regained almost their normal state at 1 week. When the BSS contained ascorbic acid, no severe damage was observed under any condition. Normal AQ-1 and ZO-1 expressions were observed even with 60 s of exposure when ascorbic acid was used.


CONCLUSIONS
A short period of irrigation of the anterior chamber with ozonated solution does not harm the corneal endothelium even when used in combination with ascorbic acid.
Suzuki
Hisaharu
H
Department of Ophthalmology, Nippon Medical School, Tokyo, Japan.
Sato
Shigeru
S
Central Institute for Electron Microscopic Research, Nippon Medical School, Tokyo, Japan.
Murano
Nao
N
Department of Ophthalmology, Nippon Medical School, Tokyo, Japan.
Matsui
Hironori
H
Department of Ophthalmology, Nippon Medical School, Tokyo, Japan.
Oharazawa
Hideaki
H
Department of Ophthalmology, Nippon Medical School, Tokyo, Japan.
Takahashi
Hiroshi
H
Department of Ophthalmology, Nippon Medical School, Tokyo, Japan. tash@nms.ac.jp.|Department of Ophthalmology, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo, 113-8603, Japan. tash@nms.ac.jp.