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


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
School of Optometry, University of Waterloo, Waterloo, ON, Canada N2L 3G1.
Hyun-Yi
HY
Youn
Kelley L
KL
Moran
Olanrewaju M
OM
Oriowo
Niels C
NC
Bols
Jacob G
JG
Sivak
D000818:Animals; D001548:Benzalkonium Compounds / Q000633:toxicity*; D001681:Biological Assay / Q000379:methods; D002417:Cattle; D003902:Detergents / Q000633:toxicity*; D005260:Female; D007908:Lens, Crystalline / Q000473:pathology*; D055095:Optics and Photonics; D012967:Sodium Dodecyl Sulfate / Q000633:toxicity*; D013501:Surface-Active Agents / Q000633:toxicity*; D046509:Tissue Culture Techniques; D014466:Ultraviolet Rays / Q000009:adverse effects*
D001548:Benzalkonium Compounds; D003902:Detergents; D013501:Surface-Active Agents; D012967:Sodium Dodecyl Sulfate