Surfactant and UV-B-induced damage of the cultured bovine lens

To evaluate in vitro methods for testing the toxicity of the surfactants, sodium dodecyl sulfate (SDS) and benzalkonium chloride (BAK), and Ultraviolet (UV)-B radiation to the bovine lens.

Lenses were dissected from bovine eyes--obtained from a local abattoir--and incubated in M199 culture medium at 37 degrees C, with 4% CO(2) and 96% air atmosphere. For the SDS and BAK experiments, the lenses (n = 153) were exposed directly to 0.001%, 0.01%, 0.1%, and 1.0% solutions for 15 min. These lenses were then rinsed five times each with saline and medium. Another group of lenses (n = 36) was irradiated with broadband UV-B at energy levels of 1.0 and 2.0 J/cm(2) (0.445 and 0.89 J/cm(2) in the biologically effective energy levels). For all of the above experiments, lens optical quality and cellular viability of lens epithelial cells were evaluated.

The analysis of optical quality, using a scanning laser in vitro assay system, of exposed lenses treated with SDS and BAK at concentrations of 0.01%, 0.1%, and 1.0%, and with UV-B at energy levels of 0.445 and 0.89 J/cm(2) showed a dose- and time-dependent increase in back vertex distance (BVD) variability, indicating loss of sharp focus in comparison with control lenses. Both 0.001% SDS and 0.001% BAK-treated lenses did not show any optical damage until 8-days after exposure. Lenses treated with 0.01% SDS showed recovery from optical damage 6-days later after exposure. Optical damage was not shown immediately for UV-B-exposed lenses. The Alamar Blue assay data for SDS, BAK and UV-B-exposed lenses, except the 0.001% SDS treated lens group, showed also dose- and time-dependent decreases in cellular viability in comparison with the control lenses, and there was no cellular recovery during the entire culture period. Lenses treated with 0.001% SDS did not show biological damage until 8-days after exposure. It appears that cellular changes appeared earlier than optical changes.

The findings suggest that cultured bovine lenses can be evaluated by assays that probe optical properties and cellular function after exposure to surfactants and UV-B irradiation, and that the optical and biological assay methods are valuable for in vitro mild ocular toxicity research.