The evaluation of cytoxicity of ocular hypotensive therapy to cultured human corneal epithelial cells

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Abstract

BACKGROUND: Corneal epithelial defect is a corneal epithelial cells’ (CEpC) disruption of various origin. In patients with corneal epithelial defect, glaucoma is a frequent concomitant disease demanding prescription of hypotensive medications, most of which containing benzalkonium chloride.

AIM: The aim of this study is to evaluate cytotoxicity of antiglaucoma drugs, as well as benzalkonium chloride (BAK), to human CEpC in vitro.

MATERIALS AND METHODS: The study was carried out on primary cultures of human CEpC. Cytotoxicity of dorzolamide, brimonidine, timolol (dilutions 1/100, 1/50, 1/20, 1/10, a 24-hour exposure) and of BAK was estimated on the model of intact epithelium (monolayer). Benzalkonium chloride was evaluated in concentrations equals to its concentration in tested ophthalmic solutions. Cytotoxicity of dorzolamide, brimonidine, timolol (dilutions 1/100, 1/20, a 48-hour exposure) was evaluated on the model of corneal epithelial defect (damage of the monolayer). The medication’s cytotoxicity was estimated by cellular changes (phase-contrast microscopy) and by the MTS-test’s results.

RESULTS: Among BAK-free medications: dorzolamide (1/50, 1/20, 1/10 dilutions), brimonidine (1/10 dilution) — induce CEpCs’ pathological changes, whereas timolol (all tested dilutions) is non-toxic. BAK-preserved drugs: dorzolamide, brimonidine, timolol (1/100, 1/50, 1/20, 1/10 dilutions) — induce CEpCs’ damage, their viability reduction, and corneal epithelial defect closure inhibition. BAK shows similar effect in tested concentrations.

CONCLUSIONS: Cytotoxicity of antiglaucoma drugs is attributed to their component — benzalkonium chloride. Administration of preserved drugs is not reasonable in eyes with corneal epithelial defect of various origin.

About the authors

Natalia V. Fisenko

M.M. Krasnov Scientific Research Institute of Eye Diseases

Email: natfisenko@mail.ru
ORCID iD: 0000-0001-7198-4498
SPIN-code: 9750-1529

MD, Cand. Sci. (Medicine)

Russian Federation, 11 A, B, Rossolimo st., Moscow, 119021

Yusef N. Yusef

M.M. Krasnov Scientific Research Institute of Eye Diseases; I.M. Sechenov First Moscow State Medical University (Sechenov University)

Email: info@eyeacademy.ru
ORCID iD: 0000-0003-4043-456X
SPIN-code: 6891-6138

MD, Dr. Sci. (Medicine), Professor

Russian Federation, Moscow; Moscow

Anastasia M. Subbot

M.M. Krasnov Scientific Research Institute of Eye Diseases

Email: kletkagb@gmail.com
ORCID iD: 0000-0002-8258-6011
SPIN-code: 3898-2570

MD, Cand. Sci. (Medicine)

Russian Federation, Moscow

Grigory A. Osipyan

M.M. Krasnov Scientific Research Institute of Eye Diseases

Author for correspondence.
Email: Gregor79@yandex.ru
ORCID iD: 0000-0002-1056-4331
SPIN-code: 1039-0470

MD, Dr. Sci. (Medicine)

Russian Federation, Moscow

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Supplementary files

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2. Fig. 1. The morphological picture of human corneal epithelial cells’ culture (model of intact epithelium) after 24-hour exposure to dorzolamide, phase-contrast microscopy, scale 50 μm. Green arrow — spaced cell junctions, blue arrow — cytoplasmic content’s condensation, white arrow — nuclear and cytoplasmic compaction, red arrow — dead cells

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3. Fig. 2. The morphological picture of human corneal epithelial cells’ culture (model of intact epithelium) after 24-hour exposure to brimonidine, phase-contrast microscopy, scale 50 μm. Blue arrow — cytoplasmic content’s condensation, yellow arrow — nuclear and cytoplasmic condensation, red arrow — dead cells

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4. Fig. 3. The morphological picture of human corneal epithelial cells’ culture (model of intact epithelium) after 24-hour exposure to timolol, phase-contrast microscopy, scale 50 μm. Yellow arrow — nuclear and cytoplasmic condensation, red arrow — dead cells

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5. Fig. 4. The morphological picture of human corneal epithelial cells’ culture (model of intact epithelium) after 24-hour exposure to benzalkonium chloride, phase-contrast microscopy, scale — 50 μm. Blue arrow — cytoplasmic content’s condensation, yellow arrow — nuclear and cytoplasmic condensation, red arrow — dead cells

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6. Fig. 5. The morphological picture of human corneal epithelial cells’ culture (model of corneal epithelial defect) after 48-hour exposure to antiglaucoma drugs, phase-contrast microscopy, scale — 100 μm. Yellow dotted line —the borders of the epithelial defect

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