title: Reconstituted human upper airway epithelium as 3-d in vitro model for nasal polyposis.
abstract: BACKGROUND
  Primary human airway epithelial cells cultured in an air-liquid interface (ALI) develop a well-differentiated epithelium. However, neither characterization of mucociliar differentiation overtime nor the inflammatory function of reconstituted nasal polyp (NP) epithelia have been described.
  
  
  OBJECTIVE
  1st) To develop and characterize the mucociliar differentiation overtime of human epithelial cells of chronic rhinosinusitis with nasal polyps (CRSwNP) in ALI culture system; 2nd) To corroborate that 3D in vitro model of NP reconstituted epithelium maintains, compared to control nasal mucosa (NM), an inflammatory function.
  
  
  METHODS
  Epithelial cells were obtained from 9 NP and 7 control NM, and differentiated in ALI culture for 28 days. Mucociliary differentiation was characterized at different times (0, 7, 14, 21, and 28 days) using ultrastructure analysis by electron microscopy; ΔNp63 (basal stem/progenitor cell), β-tubulin IV (cilia), and MUC5AC (goblet cell) expression by immunocytochemistry; and mucous (MUC5AC, MUC5B) and serous (Lactoferrin) secretion by ELISA. Inflammatory function of ALI cultures (at days 0, 14, and 28) through cytokine (IL-8, IL-1β, IL-6, IL-10, TNF-α, and IL-12p70) and chemokine (RANTES, MIG, MCP-1, IP-10, eotaxin-1, and GM-CSF) production was analysed by CBA (Cytometric Bead Array).
  
  
  RESULTS
  In both NP and control NM ALI cultures, pseudostratified epithelium with ciliated, mucus-secreting, and basal cells were observed by electron microscopy at days 14 and 28. Displaying epithelial cell re-differentation, β-tubulin IV and MUC5AC positive cells increased, while ΔNp63 positive cells decreased overtime. No significant differences were found overtime in MUC5AC, MUC5B, and lactoferrin secretions between both ALI cultures. IL-8 and GM-CSF were significantly increased in NP compared to control NM regenerated epithelia.
  
  
  CONCLUSIONS
  Reconstituted epithelia from human NP epithelial cells cultured in ALI system provides a 3D in vitro model that could be useful both for studying the role of epithelium in CRSwNP while developing new therapeutic strategies, including cell therapy, for CRSwNP.
authors.affiliation: Clinical and Experimental Respiratory Immunoallergy, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain; CIBER of Respiratory Diseases (CIBERES), Barcelona, Spain.
authors.forename: Francisco
authors.initials: F
authors.lastname: de Borja Callejas
authors.affiliation: Clinical and Experimental Respiratory Immunoallergy, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain; CIBER of Respiratory Diseases (CIBERES), Barcelona, Spain.
authors.forename: Asunción
authors.initials: A
authors.lastname: Martínez-Antón
authors.affiliation: Clinical and Experimental Respiratory Immunoallergy, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain; Rhinology Unit & Smell Clinic, ENT Department, Hospital Clínic, Universitat de Barcelona, Barcelona, Catalonia, Spain.
authors.forename: Isam
authors.initials: I
authors.lastname: Alobid
authors.affiliation: Clinical and Experimental Respiratory Immunoallergy, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain; CIBER of Respiratory Diseases (CIBERES), Barcelona, Spain.
authors.forename: Mireya
authors.initials: M
authors.lastname: Fuentes
authors.affiliation: CIBER of Respiratory Diseases (CIBERES), Barcelona, Spain.
authors.forename: Julio
authors.initials: J
authors.lastname: Cortijo
authors.affiliation: Clinical and Experimental Respiratory Immunoallergy, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain; CIBER of Respiratory Diseases (CIBERES), Barcelona, Spain; Pneumology & Respiratory Allergy Department, Hospital Clínic, Universitat de Barcelona, Barcelona, Catalonia, Spain.
authors.forename: César
authors.initials: C
authors.lastname: Picado
authors.affiliation: Clinical and Experimental Respiratory Immunoallergy, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain; CIBER of Respiratory Diseases (CIBERES), Barcelona, Spain.
authors.forename: Jordi
authors.initials: J
authors.lastname: Roca-Ferrer
authors.affiliation: Clinical and Experimental Respiratory Immunoallergy, Institut d'Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS), Barcelona, Spain; CIBER of Respiratory Diseases (CIBERES), Barcelona, Spain; Rhinology Unit & Smell Clinic, ENT Department, Hospital Clínic, Universitat de Barcelona, Barcelona, Catalonia, Spain.
authors.forename: Joaquim
authors.initials: J
authors.lastname: Mullol
mesh_terms: D002478:Cells, Cultured; D018925:Chemokines / Q000096:biosynthesis; D004847:Epithelial Cells / Q000473:pathology* / Q000648:ultrastructure; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D007781:Lactoferrin / Q000378:metabolism; D008954:Models, Biological; D009093:Mucus / Q000378:metabolism; D009297:Nasal Mucosa / Q000473:pathology* / Q000648:ultrastructure; D009298:Nasal Polyps / Q000473:pathology*; D010641:Phenotype; D013997:Time Factors
chemical_list: D018925:Chemokines; D007781:Lactoferrin