J Korean Ophthalmol Soc.  2010 Apr;51(4):588-597. 10.3341/jkos.2010.51.4.588.

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

• Kim SJ
• Kim KH
• Lee JE
• Lee JS

• KMID: 2213403
• DOI: http://doi.org/10.3341/jkos.2010.51.4.588

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

Keyword

Artificial tear; Corneal epithelial cell toxicity; LDH; MTT

MeSH Terms

Chromatin
Cytoplasm
Epithelial Cells
Eye
Humans
L-Lactate Dehydrogenase
Light
Microscopy
Ophthalmic Solutions
Organelles
Tears
Wound Healing
Chromatin
L-Lactate Dehydrogenase
Ophthalmic Solutions

Figure

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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).

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  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.

• 

  Figure 3. LDH titers of cultured corneal epithelial cells exposed in four artificial tear drops. (* p<0.05).

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  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®.

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  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®.

Reference

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