title: Bovine corneal epithelial primary cultures as an in vitro model for ophthalmic drugs studies.
abstract: In the recent years a significant development of investigations with regard to bioavailability of ocular drugs has been noticed. The corneal epithelial barrier is the main pathway for ocular penetration of topically applied ophthalmic drugs into the anterior chamber. To work out an in vitro model of bovine corneal epithelial primary cultures and exercise it for permeability research with lipophilic and hydrophilic markers, permeability coefficients estimation of the 6-carboxyfluorescein and rhodamin B was made. The corneal epithelial cultures of the 3th or 4th passage were chosen for layered culture with inserts based on the liquid-liquid interface (for the first week) and the air-liquid interface (for the two following weeks). On the 7th, 12th, 18th, 21st experiment day TER values, and on the 21st day drug permeability coefficients, were determined. The mean TER values of the 7th, 12th, 18th, 21st day of corneal epithelial culture were: 122.14, 155.14, 198.43 and; 247.43 Wcm2, respectively. The mean values of permeability coefficients on the 21st day of culture for 6-carboxyfluorescein and rhodamin B were 3.87 +/-0.10 x 10(-6)cm/s and 3.65 +/- 0.06 x 10(-6)cm/s, respectively. We state that the in vitro bovine corneal epithelial primary culture model is useful for ocular studies.
authors.affiliation: Department of Biochemistry and Clinical Chemistry, Medical University of Warsaw, Banacha St. 1, 02-097 Warszawa, Poland. gsygitowicz@poczta.onet.pl
authors.forename: Grazyna
authors.initials: G
authors.lastname: Sygitowicz
authors.forename: Danuta
authors.initials: D
authors.lastname: Zapolska-Downar
authors.forename: Magdalena
authors.initials: M
authors.lastname: Paluch
authors.forename: Tomasz
authors.initials: T
authors.lastname: Stawarski
authors.forename: Edmund
authors.initials: E
authors.lastname: Sieradzki
authors.forename: Dariusz
authors.initials: D
authors.lastname: Sitkiewicz
mesh_terms: D000375:Aging; D000465:Algorithms; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D004396:Coloring Agents; D017097:Electric Impedance; D019573:Epithelium, Corneal / Q000166:cytology* / Q000187:drug effects* / Q000254:growth & development; D005452:Fluoresceins; D009883:Ophthalmic Solutions / Q000493:pharmacokinetics*; D010539:Permeability; D012235:Rhodamines
chemical_list: D004396:Coloring Agents; D005452:Fluoresceins; D009883:Ophthalmic Solutions; D012235:Rhodamines; C024098:6-carboxyfluorescein; C029773:rhodamine B