HCE-T cell-based permeability model: A well-maintained or a highly variable barrier phenotype?

Juretić, Marina; Dukovski, Bisera Jurišić; Krtalić, Iva; Reichl, Stephan; Cetina-Čižmek, Biserka; Filipović-Grčić, Jelena; Lovrić, Jasmina; Pepić, Ivan

doi:10.1016/j.ejps.2017.03.018

Cited by 26 publications

(18 citation statements)

References 36 publications

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“…MILCEL-MTK, MatTek Corporation). 25,27,41,42 3D corneal constructs were grown using proprietary serum-free medium at standard culture conditions (SCC, 37 6 18C, 95 6 3% relative humidity, and 5 6 0.5% [vol/vol] CO 2 ) and were maintained under submerged conditions with medium contacting both the apical and basolateral sides of the tissue until the cells reached confluence. At confluence, the medium was removed from the apical side and the cells were exposed to an air-liquid interface ( Fig.…”

Section: In Vitro Reconstructed 3d Corneal Tissue Modelmentioning

confidence: 99%

“…4). 42,[44][45][46] Light Microscopy and Hematoxylin and Eosin Staining 3D corneal tissues were harvested on day 10, rinsed in PBS, fixed in 10% neutral buffered formalin for 1 hour, dehydrated in a graded series of ethanol, and embedded in paraffin. Fivemicrometer-thick hematoxylin and eosin (H&E)-stained cross sections were prepared using standard histologic techniques 47 and examined with a light microscope (Olympus BX41; Center Valley, PA, USA) or processed using an Olympus VS120 slide scanner at 310 objective with a 30.5 camera adjustment magnification.…”

Section: Transepithelial Electrical Resistance Measurementsmentioning

confidence: 99%

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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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Section: In Vitro Reconstructed 3d Corneal Tissue Modelmentioning

confidence: 99%

Section: Transepithelial Electrical Resistance Measurementsmentioning

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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“…In vitro biocompatibility study was performed using HCE-T cells (RIKEN Cell Bank, Japan). The cells were cultivated as reported previously (11,12). For in vitro biocompatibility studies two types of cell models were employed, namely 2D and 3D HCE-T cell models.…”

Section: In Vitro Biocompatibility Studymentioning

confidence: 99%

“…For cultivation of the 3D HCE-T model, Transwell ® polycarbonate membrane cell culture inserts (0.4 mm pore size, 12 mm diameter, surface area 1.12 cm 2 , Corning B.V. Life Sciences, Amsterdam, The Netherlands) were used. The 3D HCE-T model was cultivated as reported previously (11,12). Before treatment with the NE samples, the culture medium was aspirated from the basolateral side and the inserts were washed with HBSS.…”

Section: In Vitro Biocompatibility Studymentioning

confidence: 99%

“…After incubation, the test samples were removed from the apical side and the inserts were washed with HBSS, after which the inserts were transferred to a new 12-well plate with a metal plate containing 2 mL of cell culture medium in the basolateral side, while the apical side was exposed to the air-liquid interface (ALI) overnight. MTT assay was performed according to the protocol previously reported (11,14). Briefly, after removing the cell culture medium, 0.7 mL of MTT solution in the cell culture medium (1.2 mM) was added to both apical and basolateral compartment and the model was incubated during 3 hours at 37 °C.…”

Section: In Vitro Biocompatibility Studymentioning

confidence: 99%

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In vitro evaluation of stearylamine cationic nanoemulsions for improved ocular drug delivery

et al. 2019

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Oil-in-water nanoemulsions (NEs) represent one of the formulation approaches to improve eye-related bio-availability of lipophilic drugs. The potential of cationic NEs is pronounced due to the electrostatic interaction of positively charged droplets with negatively charged mucins present in the tear film, providing prolonged formulation residence at the ocular surface. The aim of this study was to develop a cationic ophthalmic NE with cationic lipid stearylamine (SA) as a carrier of a positive charge. The addition of a nonionic surfactant provided the dual electro-steric stabilization of NEs and enabled tuning of SA concentration to achieve an optimal balance between its interaction with mucins and biocompatibility. Physicochemical characterization, stability profile, in vitro mucoadhesion study and biocompatibility study employing 3D HCE-T cell-based model of corneal epithelium pointed out the NE with 0.05 % (m/m) SA as the leading formulation. Minimizing SA content while retaining droplet/mucin interactions is of great importance for efficacy and safety of future ophthalmic drug products.

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Measurement of Transcellular Transport Rates and Intracellular Drug Sequestration in the Presence of an Extracellular Concentration Gradient

Min

Rosania

2021

Methods in Pharmacology and Toxicology

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HCE-T cell-based permeability model: A well-maintained or a highly variable barrier phenotype?

Cited by 26 publications

References 36 publications

New Human Organotypic Corneal Tissue Model for Ophthalmic Drug Delivery Studies

New Human Organotypic Corneal Tissue Model for Ophthalmic Drug Delivery Studies

In vitro evaluation of stearylamine cationic nanoemulsions for improved ocular drug delivery

Measurement of Transcellular Transport Rates and Intracellular Drug Sequestration in the Presence of an Extracellular Concentration Gradient

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