Cell culture models of the human cornea - a comparative evaluation of their usefulness to determine ocular drug absorption in-vitro.
Cell culture models of the cornea are continually developed to replace the isolated animal cornea for transcorneal drug absorption studies. The aim of this study was to determine and compare epithelial tightness and permeability of currently available corneal cell culture models to avoid interlaboratory variability and to assess their usefulness for in-vitro permeation studies. Pure epithelial cell culture models (CEPI, SIRC and HCE-T cell lines), primary cultures of human corneal epithelium (HCEpiC) and the two commercially available models (RHC and Epiocular), as well as organotypic human cornea constructs (HCC, HCC-HCE-T), were investigated and data were compared with those obtained from the excised bovine cornea. Barrier properties were assessed by measurements of transepithelial electrical resistance (TEER) and permeability of three passively absorbed substances (mannitol, testosterone and timolol maleate) with different physico-chemical properties. TEER experiments revealed weak barrier functions for all of the investigated epithelial models (<or=100-200 Omega cm2), except the HCE-T cell line. Transport studies confirmed TEER results insofar that models showing low TEER values also had higher permeation rates in comparison with the excised bovine cornea. However, models based on HCE-T cells demonstrated similar barrier properties to isolated corneal tissue. The corneal models investigated in our laboratory show clear differences in epithelial barrier function. In-vitro systems comprising the HCE-T cell line seem to be most appropriate to replace excised animal cornea for assessing corneal permeability.
Reichl
Stephan
S
Institut für Pharmazeutische Technologie, Technische Universität Braunschweig, Braunschweig, Germany. S.Reichl@tu-bs.de
D000826:Animal Testing Alternatives / Q000379:methods; D000818:Animals; D002417:Cattle; D018929:Cell Culture Techniques; D002460:Cell Line; D003315:Cornea / Q000378:metabolism*; D017097:Electric Impedance; D019573:Epithelium, Corneal / Q000378:metabolism*; D006801:Humans; D008353:Mannitol / Q000493:pharmacokinetics; D008954:Models, Biological; D009883:Ophthalmic Solutions / Q000493:pharmacokinetics*; D010539:Permeability; D011817:Rabbits; D013739:Testosterone / Q000493:pharmacokinetics; D013999:Timolol / Q000493:pharmacokinetics
D009883:Ophthalmic Solutions; D008353:Mannitol; D013739:Testosterone; D013999:Timolol