J Korean Ophthalmol Soc.  2019 Dec;60(12):1176-1184. 10.3341/jkos.2019.60.12.1176.

Comparison of the Toxicity of Olopatadine Anti-allergic Ophthalmic Agents on Rabbit Conjunctival Cells

• Jeong YH
• Jang SG
• Lee JS

• KMID: 2466168
• DOI: http://doi.org/10.3341/jkos.2019.60.12.1176

Abstract

PURPOSE
To compare the in vitro toxicity of commercial olopatadine anti-allergic ophthalmic agents on cultured rabbit's conjunctival cells according to concentrations and exposure times.
METHODS
Rabbit conjunctival cells were exposed to anti-allergic olopatadine ophthalmic agents, (Patanol® [0.1% olopatadine hydrochloride; Alcon, Fort Worth, TX, USA], Pataday® [0.2% olopatadine hydrochloride; Alcon], and Pazeo® [0.7% olopatadine hydrochloride; Alcon]) at concentrations of 5%, 10%, and 15% for periods of 30 minutes, 1, 2, 3, and 6 hours, respectively. Cell proliferation and injury assays were performed using the methylthiazoltetrazolium and lactate dehydrogenase (LDH) leakage assays. We checked the composition of the three anti-allergic agents, and performed light and transmission electron microscopy to compare the morphological changes in cells.
RESULTS
The conjunctival cell proliferation was inhibited after 1 hour exposure to each olopatadine ophthalmic agent, with significant cell proliferation inhibited using 15% of each drug. The proliferation of conjunctival cells was inhibited during 6 hours of drug exposure at all concentrations of Pataday® and Pazeo®. The titer of LDH increased from 3 hours after drug exposure, but 15% Pazeo® significantly increased the LDH titer at 2 hours after drug exposure. As the concentration of the drug increased, the LDH titer also significantly increased. The cellular morphological changes of conjunctival cells were in the increasing order of Pazeo®, Pataday®, and Patanol® with a high concentration of olopatadine hydrochloride.
CONCLUSIONS
Among the anti-allergic olopatidine ophthalmic agents, higher olopatadine concentrations in the increasing order of Pazeo®, Pataday®, and Patanol® resulted in cytoplasmic damage of conjunctival cells, but there was no severe damage to the cytoplasmic or the nuclear membranes.

Keyword

Allergy; Conjunctivitis; Olopatadine hydrochloride; Toxicity

MeSH Terms

Anti-Allergic Agents
Cell Proliferation
Conjunctivitis
Cytoplasm
Hypersensitivity
In Vitro Techniques
L-Lactate Dehydrogenase
Microscopy, Electron, Transmission
Nuclear Envelope
Olopatadine Hydrochloride*
Anti-Allergic Agents
L-Lactate Dehydrogenase
Olopatadine Hydrochloride

Figure

• 

  Figure 1. Metabolic activities of rabbit's epithelial cells as determined by Methylthiazoltetrazolium assay after 30 minutes or 1, 2, 3 or 6 hours of treatment with Patanol® (0.1% olopatadine; Alcon, Fort Worth, TX, USA), Pataday® (0.2% olopatadine; Alcon) or Pazeo® (0.7% olopatadine; Alcon) in concentrations of 5, 10 and 15%. * p < 0.05 by student's t-test.

• 

  Figure 2. LDH levels started to increase rapidly after 3 hours in groups exposed to 5%, 10% and 15% Patanol® (0.1% olopatadine; Alcon, Fort Worth, TX, USA), Pataday® (0.2% olopatadine; Alcon) or Pazeo® (0.7% olopatadine; Alcon). Only 15% concentration of Pazeo® increased LDH titer after 2 hours. * p < 0.05 by analysis of LDH = lactate dehydrogenase. variance.

• 

  Figure 3. Inverted light microscopy of rabbit conjunctival cells taken after 3 hours of exposure to three agents at 10% concentration (×200). (A) Control, (B) 10% Patanol® (Alcon, Fort Worth, TX, USA), (C) 10% Pataday® (Alcon), (D) 10% Pazeo® (Alcon). Conjunctival cells of control maintained elongated, spindle-shaped appearance. But the number of cells in (B) and (C) was decreased, and (D) showed even less.

• 

  Figure 4. Transmission electron micrography of rabbit conjunctival cells taken after 3 hours of exposure to three agents at 10% concentration (bar length 2 μ m, origianl magnification, ×8,000). (A) Control, (B) 10% Patanol® (Alcon, Fort Worth, TX, USA), (C) 10% Pataday® (Alcon), (D) 10% Pazeo® (Alcon). Conjunctival cells exposed to olopatadine showed loss of microvilli, vacuole formation, and disrupted nuclear membrane.

Reference

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