Cell-based in vitro models for predicting drug permeability

Introduction: In vitro cell models have been used to predict drug permeation in early stages of drug development, since they represent an easy and reproducible method, allowing the tracking of drug absorption rate and mechanism, with an advantageous cost–benefit ratio. Such cell-based models are mainly composed of immortalized cells with an intrinsic ability to grow in a monolayer when seeded in permeable supports, maintaining their physiologic characteristics regarding epithelium cell physiology and functionality. Areas covered: This review summarizes the most important intestinal, pulmonary, nasal, vaginal, rectal, ocular and skin cell-based in vitro models for predicting the permeability of drugs. Moreover, the similitude between in vitro cell models and in vivo conditions are discussed, providing evidence that each model may provisionally resemble different drug absorption route. Expert opinion: Despite the widespread use of in vitro cell models for drug permeability and absorption evaluation purposes, a detailed study on the properties of these models and their in vitro–in vivo correlation compared with human data are required to further use in order to consider a future drug discovery optimization and clinical development.

[1]  U. Kompella,et al.  A Biodegradable Injectable Implant Sustains Systemic and Ocular Delivery of an Aldose Reductase Inhibitor and Ameliorates Biochemical Changes in a Galactose-Fed Rat Model for Diabetic Complications , 2002, Pharmaceutical Research.

[2]  S. Chong,et al.  Cell culture-based models for intestinal permeability: a critique. , 2005, Drug discovery today.

[3]  Biana Godin,et al.  Transdermal skin delivery: predictions for humans from in vivo, ex vivo and animal models. , 2007, Advanced drug delivery reviews.

[4]  Sunil A Agnihotri,et al.  Recent advances and novel strategies in pre-clinical formulation development: an overview. , 2011, Journal of controlled release : official journal of the Controlled Release Society.

[5]  J. Polli In Vitro Studies are Sometimes Better than Conventional Human Pharmacokinetic In Vivo Studies in Assessing Bioequivalence of Immediate-Release Solid Oral Dosage Forms , 2008, The AAPS Journal.

[6]  W. Rubas,et al.  Flux measurements across Caco-2 monolayers may predict transport in human large intestinal tissue. , 1996, Journal of pharmaceutical sciences.

[7]  R. Dahiya,et al.  Expression and characterization of mucins associated with the resistance to methotrexate of human colonic adenocarcinoma cell line HT29. , 1992, Cancer Research.

[8]  Thomas J. Vidmar,et al.  The Madin Darby Canine Kidney (MDCK) Epithelial Cell Monolayer as a Model Cellular Transport Barrier , 2004, Pharmaceutical Research.

[9]  Alan R. Boobis,et al.  In vitro prediction of gastrointestinal absorption and bioavailability: an experts' meeting report , 2001, European Journal of Clinical Pharmacology.

[10]  Ben Forbes,et al.  The human bronchial epithelial cell line 16HBE14o- as a model system of the airways for studying drug transport. , 2003, International journal of pharmaceutics.

[11]  Hoang Vu Dang,et al.  Nasal absorption enhancement strategies for therapeutic peptides: an in vitro study using cultured human nasal epithelium. , 2002, International journal of pharmaceutics.

[12]  Birger Brodin,et al.  Calu-3 cells grown under AIC and LCC conditions: implications for dipeptide uptake and transepithelial transport of substances. , 2011, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[13]  Y. Schneider,et al.  Development of a triculture based system for improved benefit/risk assessment in pharmacology and human food , 2011, BMC proceedings.

[14]  J. Pudney,et al.  Development of an in vitro alternative assay method for vaginal irritation. , 2011, Toxicology.

[15]  U. Hopfer,et al.  Extracellular Ca2+ directly regulates tight junctional permeability in the human cervical cell line CaSki. , 1997, The American journal of physiology.

[16]  W. Lubitz,et al.  Bacterial Ghosts as antigen and drug delivery system for ocular surface diseases: Effective internalization of Bacterial Ghosts by human conjunctival epithelial cells. , 2011, Journal of biotechnology.

[17]  G. Gorodeski Vaginal-cervical epithelial permeability decreases after menopause. , 2001, Fertility and sterility.

[18]  A. Hickey,et al.  Development of a Size-dependent Aerosol Deposition Model Utilising Human Airway Epithelial Cells for Evaluating Aerosol Drug Delivery , 2004, Alternatives to laboratory animals : ATLA.

[19]  E. Toropainen,et al.  Cell culture models of the ocular barriers. , 2005, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[20]  Y. Omidi,et al.  Ocular Drug Delivery; Impact of in vitro Cell Culture Models , 2009, Journal of ophthalmic & vision research.

[21]  D. Miller,et al.  Development of a rectal nicotine delivery system for the treatment of ulcerative colitis. , 1999, International journal of pharmaceutics.

[22]  L. Marrot,et al.  Development of genotoxicity test procedures with Episkin, a reconstructed human skin model: towards new tools for in vitro risk assessment of dermally applied compounds? , 2006, Mutation research.

[23]  Dae-Duk Kim In Vitro Cellular Models for Nasal Drug Absorption Studies , 2008 .

[24]  G. Gorodeski The Cultured Human Cervical Epithelium: A New Model for Studying Paracellular Transport , 1996, The Journal of the Society for Gynecologic Investigation: JSGI.

[25]  T. Kissel,et al.  Nasal delivery of peptides: an in vitro cell culture model for the investigation of transport and metabolism in human nasal epithelium. , 1998, Journal of controlled release : official journal of the Controlled Release Society.

[26]  Kristina Luthman,et al.  Caco-2 monolayers in experimental and theoretical predictions of drug transport1PII of original article: S0169-409X(96)00415-2. The article was originally published in Advanced Drug Delivery Reviews 22 (1996) 67–84.1 , 2001 .

[27]  Per Artursson,et al.  Effects of a new lipid-based drug delivery system on the absorption of low molecular weight heparin (Fragmin) through monolayers of human intestinal epithelial Caco-2 cells and after rectal administration to rabbits , 1994 .

[28]  G. Gorodeski Estrogen modulation of epithelial permeability in cervical-vaginal cells of premenopausal and postmenopausal women , 2007, Menopause.

[29]  R. Eckert,et al.  Cultured human ectocervical epithelial cell differentiation is regulated by the combined direct actions of sex steroids, glucocorticoids, and retinoids. , 1990, The Journal of clinical endocrinology and metabolism.

[30]  A. Minghetti,et al.  Analysis of in vitro release through reconstructed human epidermis and synthetic membranes of multi-vitamins from cosmetic formulations. , 2010, Journal of pharmaceutical and biomedical analysis.

[31]  C. Ehrhardt,et al.  16HBE14o- Human Bronchial Epithelial Cell Layers Express P-Glycoprotein, Lung Resistance-Related Protein, and Caveolin-1 , 2003, Pharmaceutical Research.

[32]  K. Leong,et al.  Transport of chitosan-DNA nanoparticles in human intestinal M-cell model versus normal intestinal enterocytes. , 2010, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[33]  A. Mitra,et al.  Barriers in Ocular Drug Delivery , 2008 .

[34]  Dae-Duk Kim,et al.  Transport of anti-allergic drugs across the passage cultured human nasal epithelial cell monolayer. , 2005, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[35]  B. Sarmento,et al.  Antioxidants in the Prevention and Treatment of Diabetic Retinopathy - A Review , 2010 .

[36]  Yves-Jacques Schneider,et al.  An improved in vitro model of human intestinal follicle-associated epithelium to study nanoparticle transport by M cells. , 2007, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[37]  Jian-ping Gao,et al.  Cytokine responses by conjunctival epithelial cells: an in vitro model of ocular inflammation. , 2008, Cytokine.

[38]  P. T. N. Sarkis,et al.  Anti-gp120 Minibody Gene Transfer to Female Genital Epithelial Cells Protects against HIV-1 Virus Challenge In Vitro , 2011, PloS one.

[39]  B. Sarmento,et al.  Chitosan-coated solid lipid nanoparticles enhance the oral absorption of insulin , 2011, Drug Delivery and Translational Research.

[40]  C. Lehr,et al.  A new Pharmaceutical Aerosol Deposition Device on Cell Cultures (PADDOCC) to evaluate pulmonary drug absorption for metered dose dry powder formulations. , 2011, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[41]  M. Donovan,et al.  Comparison of human tracheal/bronchial epithelial cell culture and bovine nasal respiratory explants for nasal drug transport studies. , 2005, Journal of pharmaceutical sciences.

[42]  P. Wertz,et al.  The human epidermis models EpiSkin, SkinEthic and EpiDerm: an evaluation of morphology and their suitability for testing phototoxicity, irritancy, corrosivity, and substance transport. , 2005, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[43]  Ben Forbes,et al.  Human respiratory epithelial cell culture for drug delivery applications. , 2005, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[44]  G. Gorodeski Aging and estrogen effects on transcervical-transvaginal epithelial permeability. , 2005, The Journal of clinical endocrinology and metabolism.

[45]  D. B. Clarke,et al.  Differential expression of organic cation transporters in normal and polyps human nasal epithelium: implications for in vitro drug delivery studies. , 2011, International journal of pharmaceutics.

[46]  Stephan Reichl,et al.  RPMI 2650 epithelial model and three-dimensional reconstructed human nasal mucosa as in vitro models for nasal permeation studies. , 2010, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[47]  G. Doncel,et al.  Preclinical evaluation of UC781 microbicide vaginal drug delivery , 2011, Drug Delivery and Translational Research.

[48]  Qiang Zhang,et al.  Transport of proteins and peptides across human cultured alveolar A549 cell monolayer. , 2004, International journal of pharmaceutics.

[49]  A. Baylor,et al.  Development of immortalized rat conjunctival epithelial cell lines: an in vitro model to examine transepithelial antigen delivery. , 2006, Experimental eye research.

[50]  R A Morrison,et al.  Current methodologies used for evaluation of intestinal permeability and absorption. , 2000, Journal of pharmacological and toxicological methods.

[51]  Peter Gehr,et al.  A three-dimensional cellular model of the human respiratory tract to study the interaction with particles. , 2005, American journal of respiratory cell and molecular biology.

[52]  R. Eckert,et al.  Maintenance of in vivo-like keratin expression, sex steroid responsiveness, and estrogen receptor expression in cultured human ectocervical epithelial cells. , 1990, Endocrinology.

[53]  H. Kuiper,et al.  Comparison of Caco-2, IEC-18 and HCEC cell lines as a model for intestinal absorption of genistein, daidzein and their glycosides. , 2004, Environmental toxicology and pharmacology.

[54]  J. J. van de Sandt,et al.  Monolayers of IEC-18 cells as an in vitro model for screening the passive transcellular and paracellular transport across the intestinal barrier: comparison of active and passive transport with the human colon carcinoma Caco-2 cell line. , 2002, Environmental toxicology and pharmacology.

[55]  J. Whittembury,et al.  A novel fluorescence chamber for the determination of volume changes in human CaSki cell cultures attached on filters , 2007, Cell Biochemistry and Biophysics.

[56]  G. Seigel,et al.  EVALUATION OF IN VITRO EFFECTS OF BEVACIZUMAB (AVASTIN) ON RETINAL PIGMENT EPITHELIAL, NEUROSENSORY RETINAL, AND MICROVASCULAR ENDOTHELIAL CELLS , 2006, Retina.

[57]  M. Brewster,et al.  The use of human nasal in vitro cell systems during drug discovery and development. , 2005, Toxicology in vitro : an international journal published in association with BIBRA.

[58]  Claude Roques,et al.  Correlation Between Oral Drug Absorption in Humans, and Apparent Drug Permeability in TC-7 Cells, A Human Epithelial Intestinal Cell Line: Comparison with the Parental Caco-2 Cell Line , 1998, Pharmaceutical Research.

[59]  K. Roemer,et al.  Differentiation of human alveolar epithelial cells in primary culture: morphological characterization and synthesis of caveolin-1 and surfactant protein-C , 2002, Cell and Tissue Research.

[60]  Claus-Michael Lehr,et al.  An in vitro triple cell co-culture model with primary cells mimicking the human alveolar epithelial barrier. , 2011, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[61]  J. Kraehenbuhl,et al.  Conversion by Peyer's patch lymphocytes of human enterocytes into M cells that transport bacteria. , 1997, Science.

[62]  R. Eckert,et al.  Human uterine cervical epithelial cells grown on permeable support--a new model for the study of differentiation. , 1994, Differentiation; research in biological diversity.

[63]  Ben Forbes,et al.  Culture of Calu-3 Cells at the Air Interface Provides a Representative Model of the Airway Epithelial Barrier , 2006, Pharmaceutical Research.

[64]  M. Romero,et al.  Characterization of Paracellular Permeability in Cultured Human Cervical Epithelium , 1994, The Journal of the Society for Gynecologic Investigation: JSGI.

[65]  Li Di,et al.  Coexistence of passive and carrier-mediated processes in drug transport , 2010, Nature Reviews Drug Discovery.

[66]  A. Barbat,et al.  Resistance to high concentrations of methotrexate and 5‐fluorouracil of differentiated HT‐29 colon‐cancer cells is restricted to cells of enterocytic phenotype , 1998, International journal of cancer.

[67]  U. Hopfer,et al.  Regulation of the Paracellular Permeability of Cultured Human Cervical Epithelium by a Nucleotide Receptor , 1995, The Journal of the Society for Gynecologic Investigation: JSGI.

[68]  Maria Jose Alonso,et al.  Chitosan-based nanostructures: a delivery platform for ocular therapeutics. , 2010, Advanced drug delivery reviews.

[69]  David J Brayden,et al.  Expression of specific markers and particle transport in a new human intestinal M-cell model. , 2000, Biochemical and biophysical research communications.

[70]  G. Amidon,et al.  HT29-MTX/Caco-2 cocultures as an in vitro model for the intestinal epithelium: in vitro-in vivo correlation with permeability data from rats and humans. , 1996, Journal of pharmaceutical sciences.

[71]  Alex Avdeef,et al.  How well can the Caco-2/Madin-Darby canine kidney models predict effective human jejunal permeability? , 2010, Journal of medicinal chemistry.

[72]  C. Ehrhardt,et al.  Drug Absorption Studies: In Situ, In Vitro and In Silico Models , 2021 .

[73]  G. Gorodeski Estrogen increases the permeability of the cultured human cervical epithelium by modulating cell deformability. , 1998, American journal of physiology. Cell physiology.

[74]  T. Shaffer,et al.  Cultured Human Airway Epithelial Cells (Calu-3): A Model of Human Respiratory Function, Structure, and Inflammatory Responses , 2010, Critical care research and practice.

[75]  C. Kirkpatrick,et al.  Primary human coculture model of alveolo-capillary unit to study mechanisms of injury to peripheral lung , 2009, Cell and Tissue Research.

[76]  B. Sarmento,et al.  Facilitated nanoscale delivery of insulin across intestinal membrane models. , 2011, International journal of pharmaceutics.

[77]  M. Wilén,et al.  Relative contribution of phosphatidylcholine and monoglyceride to absorption enhancement of low molecular weight heparin (Fragmin) by a new lipid-based drug delivery system in monolayers of human intestinal epithelial Caco-2 cells and after rectal administration to rabbits , 1994 .

[78]  P. Artursson,et al.  Co-cultures of human intestinal goblet (HT29-H) and absorptive (Caco-2) cells for studies of drug and peptide absorption , 1995 .

[79]  J. Shao,et al.  Delivery of TEM beta-lactamase by gene-transformed Lactococcus lactis subsp. lactis through cervical cell monolayer. , 2006, International journal of pharmaceutics.

[80]  Kinam Park,et al.  In vitro-in vivo correlation: perspectives on model development. , 2011, International journal of pharmaceutics.

[81]  H. Galla,et al.  Monolayers of porcine alveolar epithelial cells in primary culture as an in vitro model for drug absorption studies. , 2007, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[82]  P. Artursson,et al.  Absorption enhancement in intestinal epithelial Caco-2 monolayers by sodium caprate: assessment of molecular weight dependence and demonstration of transport routes. , 1998, Journal of drug targeting.

[83]  Hussain Fatakdawala,et al.  Hydrogen peroxide mediated transvaginal drug delivery. , 2011, International journal of pharmaceutics.

[84]  Gordon L Amidon,et al.  A Mechanistic Approach to Understanding the Factors Affecting Drug Absorption: A Review of Fundamentals , 2002, Journal of clinical pharmacology.

[85]  Zhong Zuo,et al.  An approach for rapid development of nasal delivery of analgesics--identification of relevant features, in vitro screening and in vivo verification. , 2011, International journal of pharmaceutics.

[86]  J. Goldfarb,et al.  Seminal fluid factor increases the resistance of the tight junctional complex of cultured human cervical epithelium CaSki cells. , 1998, Fertility and sterility.

[87]  K. Luthman,et al.  Caco-2 monolayers in experimental and theoretical predictions of drug transport. , 2001, Advanced drug delivery reviews.

[88]  David J Brayden,et al.  Cell culture modeling of specialized tissue: identification of genes expressed specifically by follicle-associated epithelium of Peyer's patch by expression profiling of Caco-2/Raji co-cultures. , 2004, International Immunology.

[89]  Brigitte E. Sanders-Beer,et al.  Prevention of vaginal SHIV transmission in macaques by a live recombinant Lactobacillus , 2011, Mucosal Immunology.

[90]  A. Stammati,et al.  The Caco-2 cell line as a model of the intestinal barrier: influence of cell and culture-related factors on Caco-2 cell functional characteristics , 2005, Cell Biology and Toxicology.

[91]  B. Rothen‐Rutishauser,et al.  A novel cell compatible impingement system to study in vitro drug absorption from dry powder aerosol formulations. , 2009, European journal of pharmaceutics and biopharmaceutics : official journal of Arbeitsgemeinschaft fur Pharmazeutische Verfahrenstechnik e.V.

[92]  Yong-Hae Han,et al.  Current industrial practices of assessing permeability and P-glycoprotein interaction , 2006, The AAPS Journal.

[93]  G L Amidon,et al.  Caco-2 versus Caco-2/HT29-MTX co-cultured cell lines: permeabilities via diffusion, inside- and outside-directed carrier-mediated transport. , 2000, Journal of pharmaceutical sciences.

[94]  B. Hirst,et al.  Predicting oral drug absorption and hepatobiliary clearance: Human intestinal and hepatic in vitro cell models. , 2006, Environmental toxicology and pharmacology.

[95]  V. H. Lee,et al.  The Conjunctival Barrier in Ocular Drug Delivery , 2008 .

[96]  Bruno Sarmento,et al.  Models to predict intestinal absorption of therapeutic peptides and proteins. , 2012, Current drug metabolism.

[97]  Nilesh Patel,et al.  Drug permeability in 16HBE14o- airway cell layers correlates with absorption from the isolated perfused rat lung. , 2005, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[98]  Claus-Michael Lehr,et al.  Models for Skin Absorption and Skin Toxicity Testing , 2008 .

[99]  A. Urtti,et al.  Culture model of human corneal epithelium for prediction of ocular drug absorption. , 2001, Investigative ophthalmology & visual science.

[100]  C. Ehrhardt,et al.  Drug Absorption Studies , 2008 .

[101]  C. Ehrhardt,et al.  Large Porous Particle Impingement on Lung Epithelial Cell Monolayers—Toward Improved Particle Characterization in the Lung , 2003, Pharmaceutical Research.

[102]  P. Artursson,et al.  Transport of nanoparticles across an in vitro model of the human intestinal follicle associated epithelium. , 2005, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[103]  Philippe Arnaud,et al.  Short term Caco-2/TC7 cell culture: comparison between conventional 21-d and a commercially available 3-d system. , 2004, Biological & pharmaceutical bulletin.

[104]  Young H. Kwon,et al.  Retinal synthesis and deposition of complement components induced by ocular hypertension. , 2006, Experimental eye research.

[105]  Marilyn J Aardema,et al.  Dermal penetration and metabolism of p-aminophenol and p-phenylenediamine: application of the EpiDerm human reconstructed epidermis model. , 2009, Toxicology letters.

[106]  P. Saha,et al.  Reciprocal regulation of the primary sodium absorptive pathways in rat intestinal epithelial cells. , 2011, American journal of physiology. Cell physiology.

[107]  Patric Stenberg,et al.  Transport of Lipophilic Drug Molecules in a New Mucus-Secreting Cell Culture Model Based on HT29-MTX Cells , 2001, Pharmaceutical Research.

[108]  J. Pudney,et al.  Organotypic human vaginal-ectocervical tissue model for irritation studies of spermicides, microbicides, and feminine-care products. , 2006, Toxicology in vitro : an international journal published in association with BIBRA.

[109]  Kirsten Peters,et al.  Lung epithelial cell lines in coculture with human pulmonary microvascular endothelial cells: development of an alveolo-capillary barrier in vitro , 2004, Laboratory Investigation.

[110]  G Mannens,et al.  Strategies for absorption screening in drug discovery and development. , 2001, Current topics in medicinal chemistry.

[111]  P. Annaert,et al.  In Vitro Screening Models to Assess Intestinal Drug Absorption and Metabolism , 2008 .