In vitro cell culture model for anti-cataract drug penetration studies.

Abstract

Immortalized human corneal epithelial cells (HCECs) and human lens epithelial cells (HLECs) were cultured in vitro. Cells were observed under a phase-contrast microscope and the integrity of cell monolayers was assayed by transepithelial electrical resistance (TEER) determination. The permeability of disulfiram (DSF) through a HCECs monolayer was compared with that of DSF through an excised rabbit cornea. The permeability coefficients of DSF through a HCECs monolayer and excised rabbit cornea were 29.5 +/- 4.8 x 10(-6) cm/s and 34.7 +/- 5.2 x 10(-6) cm/s, respectively. Diethyldithiocarbamate (DDC) had high permeability through HLECs monolayer with a permeability coefficient of 44.6 +/- 7.1 x 10(-6) cm/s. The cytotoxicity of DDC against HLECs was investigated using the trypan blue exclusion test. For a DDC concentration of 5 mmol/l, more than 85% cells were viable. DH3a1 mRNA was expressed in cultured HLECs. The expression of aldehyde dehydrogenase 3a1 (ALDH3a1), which may be be responsible for DSF-DDC conversion, was detected using RT-PCR and agarose gels electrophoresis. These results demonstrate that the permeability of DSF can be detected and intra-ocular drug action may be predicted using the cultured HCEC and HLEC monolayers as model.

• Authors: T Y Jiang, S L Wang, Y Liu, S F Nie, Y Ito, N Nagai, C F Wu
• Journal: Die Pharmazie (Pharmazie) Vol. 62 Issue 10 Pg. 767-72 (Oct 2007) ISSN: 0031-7144 [Print] Germany
• PMID: 18236782 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Collagen Type IV
• Coloring Agents
• Ditiocarb
• Aldehyde Dehydrogenase
• Trypan Blue
• Disulfiram

Topics

• Aldehyde Dehydrogenase (genetics)
• Animals
• Cataract (drug therapy, metabolism)
• Cell Line
• Cell Membrane Permeability
• Collagen Type IV (metabolism)
• Coloring Agents
• Cornea (metabolism)
• Disulfiram (pharmacokinetics)
• Ditiocarb (pharmacokinetics)
• Electric Conductivity
• Epithelial Cells (metabolism)
• Epithelium, Corneal (metabolism)
• Humans
• In Vitro Techniques
• Lens, Crystalline (metabolism)
• Rabbits
• Reverse Transcriptase Polymerase Chain Reaction
• Trypan Blue