Microflow-Based Device for In Vitro and Ex Vivo Drug Permeability Studies

This paper presents a novel microflow-based concept for studying the permeability of in vitro cell models or ex vivo tissues. Using the proposed concept, we demonstrate how to maintain physiologically relevant test conditions and produce highly reproducible permeability values for a range (31) of drug compounds. The apparent permeability coefficients ( Papp) showed excellent correlation (0.89) with the values from experiments performed with a conventional Ussing chamber. Additionally, the microflow-based concept produces notably more concentrated samples than the conventional Ussing chamber-based approach, despite the fact that more than 10 times smaller quantities of test compounds and biological membranes are needed in the microflow-based concept.

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Introducing an Efficient In Vitro Cornea Mimetic Model for Testing Drug Permeability

Sci ◽

10.3390/sci3030030 ◽

2021 ◽

Vol 3 (3) ◽

pp. 30

Author(s):

Agnė Žiniauskaitė ◽

Vytautas Cėpla ◽

Tadas Jelinskas ◽

Romuald Eimont ◽

Artūras Ulčinas ◽

...

Keyword(s):

T Cells ◽

Ethical Issues ◽

Ex Vivo ◽

Collagen Type I ◽

Human Cornea ◽

Type I ◽

Apparent Permeability ◽

Corneal Permeability ◽

Drug Permeability

There is a growing need for novel in vitro corneal models to replace animal-based ex vivo tests in drug permeability studies. In this study, we demonstrated a corneal mimetic that models the stromal and epithelial compartments of the human cornea. Human corneal epithelial cells (HCE-T) were grown on top of a self-supporting porcine collagen-based hydrogel. Cross-sections of the multi-layers were characterized by histological staining and immunocytochemistry of zonula oc-cludens-1 protein (ZO-1) and occludin. Furthermore, water content and bssic elastic properties of the synthetized collagen type I-based hydrogels were measured. The apparent permeability coefficient (Papp) values of a representative set of ophthalmic drugs were measured and correlated to rabbit cornea Papp values found in the literature. A multilayered structure of HCE-T cells and the expression of ZO-1 and occludin in the full thickness of the multilayer were observed. The hydrogel-based corneal model exhibited an excellent correlation to rabbit corneal permeability (r = 0.96), whereas the insert-grown HCE-T multilayer was more permeable and the correlation to the rabbit corneal permeability was lower (r = 0.89). The hydrogel-based human corneal model predicts the rabbit corneal permeability more reliably in comparison to HCE-T cells grown in inserts. This in vitro human corneal model can be successfully employed for drug permeability tests whilst avoiding ethical issues and reducing costs.

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Introducing an Efficient In Vitro Cornea Mimetic Model for Testing Drug Permeability

10.20944/preprints202105.0133.v1 ◽

2021 ◽

Author(s):

Agnė Žiniauskaitė ◽

Vytautas Cėpla ◽

Tadas Jelinskas ◽

Romuald Eimont ◽

Artūras Ulčinas ◽

...

Keyword(s):

T Cells ◽

Ethical Issues ◽

Ex Vivo ◽

Collagen Type I ◽

Human Cornea ◽

Type I ◽

Apparent Permeability ◽

Corneal Permeability ◽

Drug Permeability

There is a growing need for novel in vitro corneal models to replace animal-based ex vivo test in drug permeability studies. In this study we demonstrate a corneal mimetic that models the stromal and epithelial compartments of human cornea. Human corneal epithelial cells (HCE-T) were grown on top of a self-supporting porcine collagen-based hydrogel. Cross sections of the multilayers were characterized by histological staining and immunocytochemistry of zonula occludens-1 protein (ZO-1) and occludin. Furthermore, water content and elastic properties of the synthetized collagen type I-based hydrogels were measured. The apparent permeability coefficient (Papp) values of a representative set of ophthalmic drugs were measured and correlated to rabbit cornea Papp values found in the literature. Multilayered structure of HCE-T cells and expression of ZO-1 and occludin in full thickness of multilayer were observed. The hydrogel-based corneal model exhibited excellent correlation to rabbit corneal permeability (r=0.96), whereas insert-grown HCE-T multilayer was more permeable and the correlation to the rabbit corneal permeability was lower (r=0.89). The hydrogel-based human corneal model predicts the rabbit corneal permeability more reliably in comparison to HCE-T cells grown in inserts. This in vitro human corneal model can be successfully employed for drug permeability tests whilst avoiding ethical issues and reducing costs.

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Tissue-based in vitro and ex vivo models for vaginal permeability studies

Concepts and Models for Drug Permeability Studies ◽

10.1016/b978-0-08-100094-6.00016-x ◽

2016 ◽

pp. 273-308 ◽

Cited By ~ 1

Author(s):

Alexandra Machado ◽

José das Neves

Keyword(s):

Ex Vivo ◽

Permeability Studies

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Modeling HIV-1 Latency Using Primary CD4+ T Cells from Virally Suppressed HIV-1-Infected Individuals on Antiretroviral Therapy

Journal of Virology ◽

10.1128/jvi.02248-18 ◽

2019 ◽

Vol 93 (11) ◽

Cited By ~ 4

Author(s):

Hiroshi Takata ◽

Cari Kessing ◽

Aaron Sy ◽

Noemia Lima ◽

Julia Sciumbata ◽

...

Keyword(s):

T Cells ◽

Ex Vivo ◽

Viral Reactivation ◽

Hiv Latency ◽

Cell Models ◽

Hiv Cure ◽

Hiv Reservoirs ◽

Hiv 1

ABSTRACT The low frequency of latently HIV-infected cells in vivo limits the testing of potential HIV cure strategies using cells from successfully suppressed individuals. To date, primary cell models of latency use cells infected in vitro. Primary CD4+ T cell models carrying an individual’s endogenous HIV reservoir that recapitulate in vivo conditions of HIV latency are still outstanding. We developed a primary CD4+ T cell model of HIV latency derived from memory CD4+ T cells isolated from virally suppressed HIV-infected individuals that recapitulates HIV-1 latency and viral reactivation events. This model is based on the expansion of primary CD4+ T cells up to 300-fold in cell number. These cells reestablish a resting state without active virus production after extended culture and maintain a stable number of total HIV proviruses. The ability of these cells to respond to various classes of latency-reversing agents is similar to that of ex vivo CD4+ T cells directly isolated from blood. Importantly, viral outgrowth assays confirmed the ability of these expanded cells to produce replication-competent endogenous virus. In sum, this model recapitulates ex vivo viral reactivation conditions, captures the variability between individuals with different HIV reservoirs, and provides large numbers of cells for testing multiple agents from a single donor. The use of this novel model will allow accurate exploration of novel cure approaches aimed either at promoting viral reactivation or maintaining sustained latency. IMPORTANCE Primary cell models of HIV latency have been very useful to identify mechanisms contributing to HIV latency and to evaluate potential HIV cure strategies. However, the current models utilize in vitro infection with exogenous virus that does not fully recapitulate virus reactivation profiles of endogenous HIV in in vivo-infected CD4+ T cells. In contrast, obtaining sufficient amounts of CD4+ T cells from HIV-infected individuals to interrogate the HIV reservoir in vitro requires leukapheresis. In the model we propose here, in vitro expansion and extended culture of primary CD4+ T cells isolated from virally suppressed HIV-infected individuals enable obtaining large numbers of cells harboring endogenous latent HIV reservoirs without performing leukapheresis. This model captures the variability of HIV reservoirs seeded in different individuals and should be useful to evaluate future HIV cure strategies.

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Tissue-based in vitro and ex vivo models for blood–brain barrier permeability studies

Concepts and Models for Drug Permeability Studies ◽

10.1016/b978-0-08-100094-6.00019-5 ◽

2016 ◽

pp. 343-356

Author(s):

Malgorzata Burek ◽

Ellaine Salvador ◽

Carola Y. Förster

Keyword(s):

Blood Brain Barrier ◽

Ex Vivo ◽

Brain Barrier ◽

Barrier Permeability ◽

Blood Brain Barrier Permeability ◽

Blood Brain ◽

Brain Barrier Permeability ◽

Permeability Studies

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In Vitro and Ex Vivo Permeability Studies of Paclitaxel and Doxorubicin From Drug-Eluting Biodegradable Ureteral Stents

Journal of Pharmaceutical Sciences ◽

10.1016/j.xphs.2017.02.023 ◽

2017 ◽

Vol 106 (6) ◽

pp. 1466-1474 ◽

Cited By ~ 5

Author(s):

Alexandre A. Barros ◽

Carlos Oliveira ◽

Rui L. Reis ◽

Estevão Lima ◽

Ana Rita C. Duarte

Keyword(s):

Ex Vivo ◽

Ureteral Stents ◽

Drug Eluting ◽

Permeability Studies

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Mucin dispersions as a model for the oromucosal mucus layer in in vitro and ex vivo buccal permeability studies of small molecules

European Journal of Pharmaceutics and Biopharmaceutics ◽

10.1016/j.ejpb.2017.09.016 ◽

2017 ◽

Vol 121 ◽

pp. 121-128 ◽

Cited By ~ 10

Author(s):

Eva Marxen ◽

Mette Dalskov Mosgaard ◽

Anne Marie Lynge Pedersen ◽

Jette Jacobsen

Keyword(s):

Small Molecules ◽

Ex Vivo ◽

Mucus Layer ◽

Permeability Studies

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Tissue-based in vitro and ex vivo models for ocular permeability studies

Concepts and Models for Drug Permeability Studies ◽

10.1016/b978-0-08-100094-6.00017-1 ◽

2016 ◽

pp. 309-323

Author(s):

Christian Kölln ◽

Stephan Reichl

Keyword(s):

Ex Vivo ◽

Permeability Studies

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Drug Transport across Porcine Intestine Using an Ussing Chamber System: Regional Differences and the Effect of P-Glycoprotein and CYP3A4 Activity on Drug Absorption

Pharmaceutics ◽

10.3390/pharmaceutics11030139 ◽

2019 ◽

Vol 11 (3) ◽

pp. 139 ◽

Cited By ~ 6

Author(s):

Yvonne Arnold ◽

Julien Thorens ◽

Stéphane Bernard ◽

Yogeshvar Kalia

Keyword(s):

Drug Absorption ◽

Regional Differences ◽

Drug Transport ◽

Ussing Chamber ◽

Drug Uptake ◽

Chamber System ◽

Apparent Permeability ◽

Drug Permeability ◽

Permeability Coefficients

Drug absorption across viable porcine intestines was investigated using an Ussing chamber system. The apparent permeability coefficients, Papp,pig, were compared to the permeability coefficients determined in humans in vivo, Peff,human. Eleven drugs from the different Biopharmaceutical Classification System (BCS) categories absorbed by passive diffusion with published Peff,human values were used to test the system. The initial experiments measured Papp,pig for each drug after application in a Krebs–Bicarbonate Ringer (KBR) buffer and in biorelevant media FaSSIF V2 and FeSSIF V2, mimicking fasted and fed states. Strong sigmoidal correlations were observed between Peff,human and Papp,pig. Differences in the segmental Papp,pig of antipyrine, cimetidine and metoprolol confirmed the discrimination between drug uptake in the duodenum, jejunum and ileum (and colon); the results were in good agreement with human data in vivo. The presence of the P-gp inhibitor verapamil significantly increased Papp,pig across the ileum of the P-gp substrates cimetidine and ranitidine (p < 0.05). Clotrimazole, a potent CYP3A4 inhibitor, significantly increased Papp,pig of the CYP3A4 substrates midazolam, verapamil and tamoxifen and significantly decreased the formation of their main metabolites. In conclusion, the results showed that this is a robust technique to predict passive drug permeability under fasted and fed states, to identify regional differences in drug permeability and to demonstrate the activity of P-gp and CYP3A4.

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