Effect of age on ocular irritancy as measured with in vitro bovine lenses.

PURPOSE: This project studied the effect of age on optical quality of cultured bovine lenses exposed to a number of common surfactants and alcohol. METHODS: Lenses from calves (8-18 months) and cows (2-3 years) were isolated aseptically and studied optically for 96 h after treatment with various commercial surfactants and an alcohol. Potential eye irritancy was evaluated using a scanning laser in vitro assay system which records the change in focal characteristics (back vertex distance variability or BVDV) of the bovine lenses. Lenses were divided into a total of 14 groups. Both calf and cow lenses (a total of 257 lenses were used) were arranged into control, 0.01% BAK, 1% SDS, 1.0% Triton X-100, 100% ethanol, 10% Tween-20 and 1.0% Tween-20 treatment groups. RESULTS: The cationic surfactant BAK caused the most amount of optical change to the bovine lenses, followed by SDS, Triton X-100, ethanol and then Tween-20. There was also a significant difference in BVDV between the cow and calf groups for all the treated groups, except for Tween-20, with the calf lenses showing greater optical damage. In the case of 10% Tween-20, both cow and calf lenses show equal optical damage while at 1.0% both groups show no effect and are no different from the untreated control lenses. CONCLUSION: Younger bovine lenses are more sensitive to the surfactants and alcohol tested when compared to their older counterparts, indicating that younger eyes may be more sensitive to these chemicals. The results further indicate that age is a factor that should be taken into account in assessing ocular risk.

The lens of the eye as a focusing device and its response to stress.

The continued peripheral growth of the lens, resulting in the concentration of older tissue toward the center, has the important optical consequence of producing a lens of variable refractive index. An approach consisting of the projection of fine laser beams through excised lenses in physiological solution has been used for in vitro study of lens optical quality. By varying the separation of the incident beams and/or the wavelength characteristics of the laser used, lens refractive properties and relative transparency may be examined. In the review provided, these optical properties are correlated to lens suture anatomy, lens mitochondrial morphology and function and the function of lens heat shock proteins. In addition, lens spherical aberration is evaluated as a function of accommodation. This work can be highlighted as follows: Mammalian lens suture morphology has a direct impact on lens optical function and, while suture structure of mammalian and avian lenses are very different, they both show an age-related deterioration in morphology and focusing ability. The distribution and appearance of mitochondria of the lens epithelium and superficial fiber cells are similar in all vertebrates. Lens mitochondrial integrity is correlated to lens focusing ability, suggesting a correlation between lens optical properties and lens metabolic function. The induction of cold cataract measured optically in cultured mammalian lenses is enhanced by thermal (heat) shock and this effect is prevented by inhibiting heat shock protein production. Finally, lens accommodative function can be studied by measuring lens refractive change using a physiological model involving an intact accommodative apparatus.

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

PURPOSE: 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.

Mechanisms of ocular toxicity using the in vitro bovine lens and sodium dodecyl sulfate as a chemical model.

Previous work using the in vitro bovine lens as a model has shown a correlation between toxicity and lens optical function and showed much higher sensitivity in detecting irritancy of several surfactants at much lower concentrations than the Draize score. In the current study, cultured bovine lenses were used to study the effects of the surfactant sodium dodecyl sulfate (SDS) on lens optical properties and mitochondrial integrity. Bovine lenses were exposed to SDS (0.1 to 0.00625%) for 30 min and cultured for 24 h. Compared to controls (n = 17), loss of sharp focus was evident immediately following exposure to 0.1% SDS (n = 14, p < 0.0001). At 24 h loss of sharp focus became evident in all groups. Loss of lens transparency, significant increase in lens wet weight, and axial length were seen 24 h postexposure in lenses treated with 0.1 to 0.025% SDS. Confocal analysis 24 h postexposure showed SDS concentration-dependent decrease in number and length of the mitochondria in lens epithelial and superficial cortical fiber cells. The results of this study show a correlation between lens optical properties and metabolic function and together provide a sensitive in vitro model of ocular chemical toxicity. Results of confocal analysis suggest that the mitochondrial integrity of the superficial cortical fiber cells is most sensitive to damage caused by SDS. The results further suggest that recovery of lens metabolic function is necessary for the recovery of lens optical properties.