Sensitive and Real-Time Method for Evaluating Corneal Barrier Considering Tear Flow

Nakamura, Tadahiro; Teshima, Mugen; Kitahara, Takashi; Sasaki, Hidetada; Uematsu, Masafumi; Kitaoka, Takashi; Nakashima, Makoto; Nishida, Koyo; Nakamura, Junzo; Higuchi, Shun

doi:10.1248/bpb.33.107

Cited by 16 publications

(19 citation statements)

References 19 publications

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“…Until recently, absorption studies were commonly performed in laboratory animals or in excised rabbit, bovine, or porcine corneas mounted in diffusion chambers, such as Franz diffusion cells or Ussing chambers. 9,21,22 Not only are these studies labor intensive, expensive, poorly standardized, and impractical for large-scale and rapid drug formulation screening; they also exhibit interspecies variation due to the differences in anatomy and morphology of the eyes. 9,23 Pig eyes are structurally the most similar to human eyes in terms of the size, corneal thickness, and the presence of Bowman's layer.…”

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confidence: 99%

New Human Organotypic Corneal Tissue Model for Ophthalmic Drug Delivery Studies

Kaluzhny

Kinuthia

Truong

et al. 2018

Invest. Ophthalmol. Vis. Sci.

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PURPOSE. The purpose of the current work was to develop a physiologically relevant, in vitro human three-dimensional (3D) corneal epithelial tissue model for use in ophthalmic drug development.METHODS. Normal human corneal epithelial cells were cultured at the air-liquid interface to produce the 3D corneal tissue model. Corneal barrier was determined by measuring transepithelial electrical resistance (TEER). Quantitative PCR arrays were utilized to investigate expression of 84 phase I/II metabolizing enzymes and 84 drug transporter genes. Permeability was evaluated using model compounds with a wide range of hydrophobicity, molecular weight, and excipients. Finally, different formulations of latanoprost and bimatoprost were administered and drug absorption and tissue viability and integrity were investigated.RESULTS. Histologic assessment and TEER of the corneal tissue model revealed tissue structure, thickness, and barrier formation (1000 6 146 XÁcm 2 ) comparable to native human corneal epithelium. The 3D corneal tissue expressed tight junctions, mucins, and key corneal epithelial detoxification enzymes. Drug-metabolizing enzyme and transporter gene expression in 3D corneal tissue and excised human corneal epithelium were highly correlated (r 2 ¼ 0.87). Coefficients of permeation for model drugs in the tissue model and excised rabbit corneas also showed a high correlation (r 2 ¼ 0.94). As expected, latanoprost and bimatoprost free acids had much lower permeability (Papp ¼ 1.2 3 10 À6 and 1.9 3 10 À6 ) than the corresponding prodrugs (Papp ¼ 2.5 3 10 À5 and 5.6 3 10 À5 ), respectively. The presence of 0.02% benzalkonium chloride in ophthalmic formulations significantly affected tissue barrier and viability.CONCLUSIONS. The newly developed 3D corneal tissue model appears to be very useful for evaluation of corneal drug permeability and safety during ophthalmic drug development.

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confidence: 99%

New Human Organotypic Corneal Tissue Model for Ophthalmic Drug Delivery Studies

Kaluzhny

Kinuthia

Truong

et al. 2018

Invest. Ophthalmol. Vis. Sci.

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“…19 The lower corneal TER values are indicative of the penetration of greater amounts of electrical current through the damaged superficial cells and tight junctions that exist in the epithelium. 6,7,14,15,24 In addition, these electrical changes can be measured continuously and quantitatively. Therefore, measurement of corneal TER is a suitable method for evaluating corneal permeability and irritancy.…”

Section: Discussionmentioning

confidence: 99%

“…16,17 We previously developed that new electrophysiological method mimicking tear flow in which the precorneal solution was exchanged for a buffer with a peristaltic pump at the rate of human tear flow and excised rabbit corneas. 14 The method for evaluating eyedrop-induced corneal toxicity considering tear flow is invaluable, as tears dilute the eyedrops immediately after instillation. By using this method, we were able to determine that the TER of the excised rabbit corneas was *1 k-ohm$cm 2 .…”

Section: Discussionmentioning

confidence: 99%

“…14,15 The electrophysiological experiments were performed using a Ussing chamber CHM1 (World Precision Instruments, Sarasota, FL) with a donorphase turnover system to mimic the rate of human tear turnover at 16%/min. 16,17 Ag/AgCl half-cells were screwed into short tubes, which were plugged firmly into place in the chamber luer ports.…”

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“…Photographs (A, B)and schematic illustration (C) of electrophysiological device, and equivalent circuit model for corneal epithelium (D). Modified from Ref 14. .…”

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confidence: 99%

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Investigation of Protective Effects of Sodium Hyaluronate Eyedrop Against Corneal Epithelial Disorders Using an Electrophysiological Method

Araki

Nakashima²,

Teshima³

et al. 2012

Journal of Ocular Pharmacology and Therapeutics

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In vitro reconstructed 3D corneal tissue models for ocular toxicology and ophthalmic drug development

Kaluzhny

Klausner

2021

In Vitro Cell.Dev.Biol.-Animal

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Testing of all manufactured products and their ingredients for eye irritation is a regulatory requirement. In the last two decades, the development of alternatives to the in vivo Draize eye irritation test method has substantially advanced due to the improvements in primary cell isolation, cell culture techniques, and media, which have led to improved in vitro corneal tissue models and test methods. Most in vitro models for ocular toxicology attempt to reproduce the corneal epithelial tissue which consists of 4-5 layers of non-keratinized corneal epithelial cells that form tight junctions, thereby limiting the penetration of chemicals, xenobiotics, and pharmaceuticals. Also, significant efforts have been directed toward the development of more complex threedimensional (3D) equivalents to study wound healing, drug permeation, and bioavailability. This review focuses on in vitro reconstructed 3D corneal tissue models and their utilization in ocular toxicology as well as their application to pharmacology and ophthalmic research. Current human 3D corneal epithelial cell culture models have replaced in vivo animal eye irritation tests for many applications, and substantial validation efforts are in progress to verify and approve alternative eye irritation tests for widespread use. The validation of drug absorption models and further development of models and test methods for many ophthalmic and ocular disease applications is required.

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Sensitive and Real-Time Method for Evaluating Corneal Barrier Considering Tear Flow

Cited by 16 publications

References 19 publications

New Human Organotypic Corneal Tissue Model for Ophthalmic Drug Delivery Studies

New Human Organotypic Corneal Tissue Model for Ophthalmic Drug Delivery Studies

Investigation of Protective Effects of Sodium Hyaluronate Eyedrop Against Corneal Epithelial Disorders Using an Electrophysiological Method

In vitro reconstructed 3D corneal tissue models for ocular toxicology and ophthalmic drug development

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