title: Effect of benzalkonium chloride on trabecular meshwork cells in a new in vitro 3D trabecular meshwork model for glaucoma.
abstract: OBJECTIVE
  To validate a new culture model of primary human trabecular meshwork cells (p-hTMCs) using Matrigel®, in order to mimic in vitro 3D-TM organization, and to investigate the proinflammatory effect of benzalkonium chloride (BAK) in 3D p-hTMC cultures.
  
  
  METHODS
  p-hTMCs, seeded onto Matrigel®-coated inserts were stimulated with BAK (10-4%), dexamethasone (DEX) (10-6M) or transforming growth factor-beta 2 (TGF-β2) (5ng/ml) for 48h and observed with confocal microscopy. The BAK effect at 10-4% or 5.10-3% on the gene expressions of interleukin-6 (IL-6), interleukin-8 (IL-8) and matrix metalloproteinase (MMP-9) was investigated using qRT-PCR in 2D and 3D p-hTMC cultures.
  
  
  RESULTS
  p-hTMCs seeded in Matrigel® were able to organize themselves in a 3D-spatial conformation in the different conditions tested with cross-linked actin network (CLAN) formation in presence of DEX or TGF-β2 and intercellular space contraction with TGF-β2. IL-6 and IL-8 gene expressions increased in presence of BAK in 2D and in 3D p-hTMC cultures. BAK 10-4% only showed a tendency to stimulate MMP-9 expression in p-hTMCs after 24h-recovery.
  
  
  CONCLUSIONS
  We investigated this new 3D-TM in vitro model in Matrigel®matrix for pathophysiological and toxicological purposes. It appears as a new promising tool for a better understanding of TM behavior in physiological and stress conditions, as well as toxicological evaluations of antiglaucoma eyedrops and preservatives.
authors.affiliation: Research Laboratory LR99ES11, Department of Biochemistry, Rabta Hospital, Tunis, Tunisia. Electronic address: bouchemimeryem@yahoo.fr.
authors.forename: Meryem
authors.initials: M
authors.lastname: Bouchemi
authors.affiliation: INSERM, U968, Paris F-75012, France; UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris F-75012, France; CNRS, UMR_7210, Paris F-75012, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Roubeix
authors.affiliation: INSERM, U968, Paris F-75012, France; UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris F-75012, France; CNRS, UMR_7210, Paris F-75012, France.
authors.forename: Karima
authors.initials: K
authors.lastname: Kessal
authors.affiliation: INSERM, U968, Paris F-75012, France; UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris F-75012, France; CNRS, UMR_7210, Paris F-75012, France.
authors.forename: Luisa
authors.initials: L
authors.lastname: Riancho
authors.affiliation: INSERM, U968, Paris F-75012, France; UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris F-75012, France; CNRS, UMR_7210, Paris F-75012, France.
authors.forename: Anne-Laure
authors.initials: AL
authors.lastname: Raveu
authors.affiliation: Research Laboratory LR99ES11, Department of Biochemistry, Rabta Hospital, Tunis, Tunisia; El Manar University, Higher Institute of Medical Technologies, Tunis, Tunisia.
authors.forename: Hayet
authors.initials: H
authors.lastname: Soualmia
authors.affiliation: INSERM, U968, Paris F-75012, France; UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris F-75012, France; CNRS, UMR_7210, Paris F-75012, France; Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, INSERM-DHOS, CIC 503, Paris F-75012, France; Hop Ambroise Pare, AP HP, Dept Ophthalmology, F-92100 Boulogne, France; Univ Versailles St Quentin En Yvelines, F-78180 Montigny-Le-Bretonneux, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Baudouin
authors.affiliation: INSERM, U968, Paris F-75012, France; UPMC Univ Paris 06, UMR_S 968, Institut de la Vision, Paris F-75012, France; CNRS, UMR_7210, Paris F-75012, France; Centre Hospitalier National d'Ophtalmologie des Quinze-Vingts, INSERM-DHOS, CIC 503, Paris F-75012, France; Univ Paris Descartes, Sorbonne Paris Cité, Paris F-75006, France; Faculté de Pharmacie de Paris, Univ Paris Descartes, Sorbonne Paris Cité, Paris, 17 F-75006, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Brignole-Baudouin
mesh_terms: D001548:Benzalkonium Compounds / Q000633:toxicity*; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D003094:Collagen; D003599:Cytoskeleton / Q000187:drug effects; D003907:Dexamethasone / Q000633:toxicity; D004338:Drug Combinations; D015870:Gene Expression / Q000187:drug effects; D005901:Glaucoma; D006801:Humans; D015850:Interleukin-6 / Q000235:genetics; D016209:Interleukin-8 / Q000235:genetics; D007797:Laminin; D020780:Matrix Metalloproteinase 9 / Q000235:genetics; D009883:Ophthalmic Solutions / Q000633:toxicity*; D011310:Preservatives, Pharmaceutical / Q000633:toxicity*; D011509:Proteoglycans; D014129:Trabecular Meshwork / Q000166:cytology*; D053781:Transforming Growth Factor beta2 / Q000633:toxicity
chemical_list: D001548:Benzalkonium Compounds; C508606:CXCL8 protein, human; D004338:Drug Combinations; C508600:IL6 protein, human; D015850:Interleukin-6; D016209:Interleukin-8; D007797:Laminin; D009883:Ophthalmic Solutions; D011310:Preservatives, Pharmaceutical; D011509:Proteoglycans; D053781:Transforming Growth Factor beta2; C059485:matrigel; D003907:Dexamethasone; D003094:Collagen; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9
keywords: Benzalkonium chloride; Interleukin-6; Interleukin-8; Matrigel®; Three-dimensional culture; Trabecular meshwork