RPMI 2650 epithelial model and three-dimensional reconstructed human nasal mucosa as in vitro models for nasal permeation studies.

The purpose of this study was to investigate the human nasal epithelial cell line RPMI 2650 regarding its usefulness as in vitro model for drug permeation studies. Particularly, the influence of the air-liquid interface in culture and coculture with human nasal fibroblasts (HNF) on the differentiation and permeation barrier properties of the cell layer was examined. In addition to a non-contact coculture, we developed a three-dimensional construct of the human nasal mucosa composed of a collagen matrix with embedded HNF, covered by a RPMI 2650 epithelial cell layer. Microscopic examination as well as measurement of the transepithelial electrical resistance and permeation experiments showed the importance of cultivation at the air-liquid interface. Permeation studies were performed using a paracellular marker (sodium fluorescein), a transcellular marker (propranolol-HCl) and a model substance with high molecular weight (FITC-dextran, MW 4000). The epithelial model showed an organotypic permeation barrier for paracellular, transcellular and high MW permeation. Three-dimensional reconstructed human nasal mucosa showed four- to fivefold higher permeation coefficients. Regardless of the limits of these models, both offer promise to evaluate passive drug permeation through the nasal mucosa.

[1]  Carsten Kneuer,et al.  Influence of apical fluid volume on the development of functional intercellular junctions in the human epithelial cell line 16HBE14o–: implications for the use of this cell line as an in vitro model for bronchial drug absorption studies , 2002, Cell and Tissue Research.

[2]  H. Merkle,et al.  In vitro methodologies to study nasal delivery using excised mucosa , 2002 .

[3]  Yuh-Shyan Chen,et al.  Expression of Toll-like receptors in cultured nasal epithelial cells , 2007, Acta oto-laryngologica.

[4]  Min-Ki Lee,et al.  Serially Passaged Human Nasal Epithelial Cell Monolayer for in Vitro Drug Transport Studies , 2003, Pharmaceutical Research.

[5]  J. Vonderscher,et al.  Evaluation of the human cell line RPMI 2650 as an in vitro nasal model. , 1995, Rhinology.

[6]  Selcan Türker,et al.  Nasal route and drug delivery systems , 2004, Pharmacy World and Science.

[7]  W. Rubas,et al.  Validation of excised bovine nasal mucosa as in vitro model to study drug transport and metabolic pathways in nasal epithelium. , 2000, Journal of pharmaceutical sciences.

[8]  S. Suter,et al.  Long-term cultures of polarized airway epithelial cells from patients with cystic fibrosis. , 2006, American journal of respiratory cell and molecular biology.

[9]  Peter,et al.  In vitro cell models to study nasal mucosal permeability and metabolism. , 1998, Advanced drug delivery reviews.

[10]  T. Nagai,et al.  Influence of bile salts on the permeability of insulin through the nasal mucosa of rabbits in comparison with dextran derivatives , 1991 .

[11]  Romeo,et al.  Effects of physicochemical properties and other factors on systemic nasal drug delivery. , 1998, Advanced drug delivery reviews.

[12]  F. Ahsan,et al.  Evaluation of human nasal RPMI 2650 cells grown at an air-liquid interface as a model for nasal drug transport studies. , 2008, Journal of pharmaceutical sciences.

[13]  G. Moore,et al.  Studies of a human tumor cell line with a diploid karyotype , 1964, Cancer.

[14]  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.

[15]  O. Camber,et al.  Nasal drug delivery--evaluation of an in vitro model using porcine nasal mucosa. , 1999, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[16]  R. Agu,et al.  In Situ and Ex Vivo Nasal Models for Preclinical Drug Development Studies , 2008 .

[17]  O. Camber,et al.  Permeability of porcine nasal mucosa correlated with human nasal absorption. , 2003, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[18]  R. Deterding,et al.  Acidic fibroblast growth factor and keratinocyte growth factor stimulate fetal rat pulmonary epithelial growth. , 1996, The American journal of physiology.

[19]  T. Kissel,et al.  Development of a Human Nasal Epithelial Cell Culture Model and Its Suitability for Transport and Metabolism Studies Under in Vitro Conditions , 1995, Pharmaceutical Research.

[20]  S. Lakshminrusimha,et al.  Growth factors in lung development. , 2005, Advances in clinical chemistry.

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

[22]  M. Osanai,et al.  Changes of Gap and Tight Junctions during Differentiation of Human Nasal Epithelial Cells Using Primary Human Nasal Epithelial Cells and Primary Human Nasal Fibroblast Cells in a Noncontact Coculture System , 2007, Journal of Membrane Biology.

[23]  V. Lipipun,et al.  Spray-dried mucoadhesive microspheres: Preparation and transport through nasal cell monolayer , 2006, AAPS PharmSciTech.

[24]  Ching‐Hwa Tsai,et al.  Increased Communication Among Nasal Epithelial Cells in Air‐Liquid Interface Culture , 2007, The Laryngoscope.

[25]  J. Bernaudin,et al.  Evaluation of MRP1-5 Gene Expression in Cystic Fibrosis Patients Homozygous for the ΔF508 Mutation , 2003, Pediatric Research.

[26]  P. Augustijns,et al.  In‐vitro nasal drug delivery studies: comparison of derivatised, fibrillar and polymerised collagen matrix‐based human nasal primary culture systems for nasal drug delivery studies , 2001, The Journal of pharmacy and pharmacology.

[27]  D. Wermeling,et al.  Intranasal Delivery of Recombinant Human Parathyroid Hormone [hPTH (1–34)], Teriparatide in Rats , 2004, Endocrine research.

[28]  H. Natsume,et al.  In vitro permeation of several model drugs across rabbit nasal mucosa , 1994 .

[29]  Y. Sakakura,et al.  Macromolecular permeability of the tight junction of the human nasal mucosa. , 1985, Rhinology.

[30]  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.

[31]  Y. Maitani,et al.  The enhancing effect of nasal absorption of FITC-dextran 4,400 by β-sitosterol β-d-glucoside in rabbits , 2002 .

[32]  Dahl,et al.  Anatomy, physiology and function of the nasal cavities in health and disease. , 1998, Advanced drug delivery reviews.

[33]  V. H. Lee,et al.  Monolayers of Human Alveolar Epithelial Cells in Primary Culture for Pulmonary Absorption and Transport Studies , 1999, Pharmaceutical Research.

[34]  M. Knowles,et al.  Abnormal apical cell membrane in cystic fibrosis respiratory epithelium. An in vitro electrophysiologic analysis. , 1987, The Journal of clinical investigation.

[35]  S. Quay,et al.  Therapeutic utility of a novel tight junction modulating peptide for enhancing intranasal drug delivery. , 2006, Journal of pharmaceutical sciences.

[36]  D. Wall,et al.  Permeability Characteristics of Calu-3 Human Bronchial Epithelial Cells: In Vitro - In Vivo Correlation to Predict Lung Absorption in Rats , 2002, Journal of drug targeting.

[37]  L. Illum,et al.  Intranasal delivery: physicochemical and therapeutic aspects. , 2007, International journal of pharmaceutics.

[38]  C. V. van Blitterswijk,et al.  Effect of fibroblasts on epidermal regeneration , 2002, The British journal of dermatology.

[39]  S. Quay,et al.  Development of a novel high-concentration galantamine formulation suitable for intranasal delivery. , 2005, Journal of pharmaceutical sciences.

[40]  J. Bernaudin,et al.  CFTR, MDR1, and MRP1 Immunolocalization in Normal Human Nasal Respiratory Mucosa , 2000, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[41]  Min-Ki Lee,et al.  Air-Liquid Interface Culture of Serially Passaged Human Nasal Epithelial Cell Monolayer for In Vitro Drug Transport Studies , 2005, Drug delivery.

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

[43]  L. Ostrowski,et al.  Culture of murine nasal epithelia: model for cystic fibrosis. , 2006, American journal of physiology. Lung cellular and molecular physiology.

[44]  T. Kissel,et al.  In-vitro Cell Culture Models of the Nasal Epithelium: A Comparative Histochemical Investigation of Their Suitability for Drug Transport Studies , 1996, Pharmaceutical Research.

[45]  R. Caldwell,et al.  Serum Opens Tight Junctions and Reduces ZO‐1 Protein in Retinal Epithelial Cells , 1997, Journal of neurochemistry.

[46]  E Bell,et al.  Living tissue formed in vitro and accepted as skin-equivalent tissue of full thickness. , 1981, Science.

[47]  M I Ugwoke,et al.  The biopharmaceutical aspects of nasal mucoadhesive drug delivery , 2001, The Journal of pharmacy and pharmacology.

[48]  R A Thoft,et al.  Basement membrane synthesis by human corneal epithelial cells in vitro. , 1994, Investigative ophthalmology & visual science.

[49]  S. Werner,et al.  Paracrine regulation of keratinocyte proliferation and differentiation. , 2001, Trends in cell biology.

[50]  P. Flint,et al.  Coculture of mesenchymal stem cells and respiratory epithelial cells to engineer a human composite respiratory mucosa. , 2004, Tissue engineering.