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Kim, Kim, Lee, and Lee: Effect of Artificial Tears Used in Contact Lens-wearing Eyes on Human Corneal Epithelial Cells in Vitro
Original Article

J Korean Ophthalmol Soc 2010;51(4):588-597.

Published online: 25 September 2010

DOI: https://doi.org/10.3341/jkos.2010.51.4.588

Effect of Artificial Tears Used in Contact Lens-wearing Eyes on Human Corneal Epithelial Cells in Vitro

Su Jin Kim, MD, Kyong Ho Kim, MD, Ji Eun Lee, MD, PhD, Jong Soo Lee, MD, PhD

Department of Ophthalmology, Medical Research Institute, Pusan National University College of Medicine, Busan, Korea

Address reprint requests to Jong Soo Lee, MD, PhD Department of Ophthalmology, Pusan National University Hospital #1-10 Ami-dong, Seo-gu, Busan 602-739, Korea Tel: 82-51-240-7323, Fax: 82-51-242-7341, E-mail: jongsool@pusan.ac.kr

Received 2 July 2009       Accepted 4 February 2010

Copyright © 2010 Korean Ophthalmological Society

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

To investigate the biologic effects of topical ocular artificial tears used in patients wearing contact lens on in vitro corneal epithelial cells.

Methods

The efficacies of the topical artificial tears Iris®, Irisplus®, Eyemiru contact pure®, and Eye2O® were evaluated using the MTT and wound healing assays. Cell damage was determined using lactate dehydrogenase (LDH), and the solution ingredients were analyzed. Cellular morphologies were examined by inverted light microscopy and transmission electromicroscopy.

Results

Metabolic activity of corneal epithelial cells, as determined by the MTT assay, decreased in the Iris® eye drop group, but those of the other groups were similar to that of the control. The LDH titers increased up to one hour after Iris® eye drop use, and the increased level was maintained for 24 hours. The other three artificial tears showed similar low LDH titers to that of the control. Cellular migration was not observed, although cellular damage to the corneal epithelial cells, such as chromatin margination and cytoplasmic organelle swelling, was prominent with Iris® use.

Conclusions

Among four brands of topical artificial tear drops used among patients wearing contact lens, Iris® caused markedly more severe damage to cultured human corneal epithelial cells than did Irisplus®, Eyemiru contact pure®, or Eye2O®.

Keywords: Artificial tear, Corneal epithelial cell toxicity, LDH, MTT

References

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Figure 1.
The absorption rate of the water-insolubale formazan dye in corneal epithelial cell exposed in four artificial tear drops by a scanning spectrometer (ELISA reader) (* p<0.05).
jkos-51-588f1.tif
Figure 2.
Scratch assay of corneal epithelial cells after 0-hour exposure to (A1) control (B1) 20% Iris®, (C1) Irisplus®, (D1) Eyemiru contactpure®, (E1) Eye2O®, 7-hour exposure to (A2) control (B2) 20% Iris®, (C2) Irisplus®, (D2) Eyemiru contactpure®, (E2) Eye2O®, 24-hour exposure to (A3) control (B3) 20% Iris®, (C3) Irisplus®, (D3) Eyemiru contactpure® and (E3) Eye2O®. In control group (A), normal corneal epithelial cells were proliferated and moved into scratched scar in the culture medium. After exposure to Iris® (B), corneal epithelial cells were not proliferated and moved into scratched scar in the culture medium.
jkos-51-588f2.tif
Figure 3.
LDH titers of cultured corneal epithelial cells exposed in four artificial tear drops. (* p<0.05).
jkos-51-588f3.tif
Figure 4.
Inverted light micrographs of corneal epithelial cells after 4-hour exposure to (A) control, (B) Iris®, (C) Irisplus®, (D) Eyemiru contactpure®, and (E) Eye2O®.
jkos-51-588f4.tif
Figure 5.
Transmission electron micrographs of corneal epithelial cells appeared after 4-hour exposure to (A) control, (B) 20% Iris®, (C) 20% Irisplus®, (D) 20% Eyemiru contactpure®, and (E) 20% Eye2O®. (bar length 2 um, original magnification, ×2000∼4000). In general, the plasma membrane with microvilli (black arrow head), nuclear membrane, and nuclei of conjunctival cells were visible. Iris® had more severe and damaged cellular structures, such as the plasma membranes with microvilli being disrupted (white arrow head), well-developed vacuole formation (black arrow), and chromatin margination of the nucleus (white arrow), rather than IrisPlus®, Eyemiru contactpure®, Eye2O®.
jkos-51-588f5.tif
Table 1.
The ingredient, electrolyte composition, pH, osmolarity and preservatives of the commercial four artificial tear drops
Artificial tear Chief ingredient Na+(mEq/L) K+(mEq/L) Cl-(mEq/L) pH Osmolarity(mosm/kg) Preservative(mg/ml)
Iris® Hydroxypropylmethylcellulose 122.4 15.97 134.1 5.0 296 BAC 0.1
Irisplus® Chondroitin sodium sulfate, 91.4 9.19 87.2 6.5 362 0
  hydroxypropylmethylcellulose            
Eyemiru Potassium 1-aspartate, aminoethyl 71.1 25.47 53.9 7.0 392 0
   contactpure®    sulfonic acid, chondroitin sodium sulffate            
Eye2O® Povidone 28.7 19.36 46.2 5.0 320 0
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