Development of cultured rabbit corneal epithelium for drug permeation studies: a comparison with excised rabbit cornea.
The aim of this study was to prepare and test an artificial corneal epithelium (reconstituted rabbit corneal epithelium, RRCE) exhibiting barrier characteristics and paracellular permeability similar to those of native rabbit cornea. The RRCE was obtained from a rabbit corneal epithelium (RCE) cell line grown for 8 days in submerged culture, then for 7 days in air-interface conditions on Snapwell polyester membranes. Permeation studies on the RRCE were carried out in comparison with rabbit excised corneas in vitro, using timolol maleate (TM) as the test drug, alone and in association with the following ocular permeation enhancers: benzalkonium chloride, ethylene-diaminetetraacetic acid sodium salt, polyethoxylated castor oil, polyoxyethylene stearyl ether, sodium deoxycholate, and escin. The integrity of the RRCE was assessed by measuring the transepithelial electrical resistance (TEER) during culture time and after every permeation experiment. When TM was tested alone, the permeation parameters (apparent permeability coefficient, lag time) obtained with the RRCE were similar to those of excised rabbit corneas. The artificial epithelium, however, was less sensitive than native cornea to the effect of permeation enhancers.
Burgalassi
Susi
S
Department of Bioorganic Chemistry and Biopharmaceutics, University of Pisa, Pisa, Italy. burgal@farm.unipi.it
Monti
Daniela
D
Brignoccoli
Annalisa
A
Fabiani
Ortenzio
O
Lenzi
Carla
C
Pirone
Andrea
A
Chetoni
Patrizia
P
D000277:Adjuvants, Pharmaceutic / Q000494:pharmacology; D000818:Animals; D000959:Antihypertensive Agents / Q000493:pharmacokinetics; D001548:Benzalkonium Compounds / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D003315:Cornea / Q000378:metabolism* / Q000601:surgery; D004305:Dose-Response Relationship, Drug; D017097:Electric Impedance; D019573:Epithelium, Corneal / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D004928:Escin / Q000494:pharmacology; D008297:Male; D046529:Microscopy, Electron, Transmission; D010539:Permeability; D011817:Rabbits; D013997:Time Factors; D013999:Timolol / Q000493:pharmacokinetics*
D000277:Adjuvants, Pharmaceutic; D000959:Antihypertensive Agents; D001548:Benzalkonium Compounds; D004928:Escin; D013999:Timolol