RESUMO
Purpose: Interactions between tear proteins and the interfaces of contact lenses can be complex and can influence contact lens wear success. Tear proteins, including lysozyme, function to maintain the balance of ocular surface homeostasis, as evidenced by the effects of its conformation relative to stabilizing the tear film and its potential impact on corneal epithelial cells. Contact lens manufacturers include components in lens care and blister package solutions to help stabilize the tear film and preserve homeostasis. This in vitro study was performed to evaluate the ability of daily disposable contact lens package solutions to stabilize lysozyme and preserve its native conformation under denaturing conditions. Methods: Lysozyme was added to contact lens solutions sampled from kalifilcon A, etafilcon A, senofilcon A, narafilcon A, nelfilcon A, verofilcon A, delefilcon A, somofilcon A, and stenfilcon A blister packages, then mixed with the protein denaturant sodium lauryl sulfate. Lysozyme activity was evaluated by adding test solutions to a suspension of Micrococcus luteus. Native lysozyme lyses the Micrococcus luteus cell wall, which decreases suspension turbidity. Stabilization of lysozyme activity was determined by comparing suspension turbidity before and after exposure to test solutions. Results: Lysozyme stabilization was 90.7% for kalifilcon A solution, a statistically significant improvement (p < 0.05) compared to phosphate buffered saline (PBS, negative control). No significant improvement was observed with any other contact lens solution (all lysozyme stabilization < 5.00%). Conclusion: The representative tear protein lysozyme was significantly more stable in the novel kalifilcon A contact lens solution containing multiple moisturizers and osmoprotectants than in PBS or other daily disposable contact lens solutions. The lysozyme activity assay provides mechanistic evidence that the kalifilcon A contact lens solution can stabilize proteins under conditions that typically denature proteins, which may contribute to maintaining ocular surface homeostasis.
RESUMO
The range and extent of tear proteins removed by various multi-purpose solutions has been investigated, but there is little information in the literature about their ability to prevent denaturation of tear proteins, particularly lysozyme. The purpose of this study was to determine the ability of Bausch+Lomb Biotrue™ multi-purpose solution and other care solutions to affect denaturation of lysozyme using a lysozyme activity assay. The test solutions used were: Biotrue multi-purpose solution, Bausch+Lomb renu(®) fresh™, formerly ReNu MultiPlus(®), Alcon OPTI-FREE RepleniSH, Alcon OPTI-FREE EXPRESS, CIBA VISION AQuify, and AMO COMPLETE Multi-Purpose Solution Easy Rub Formula. A phosphate-buffered saline (PBS) solution served as a control. The test and control solutions containing lysozyme were exposed to sodium dodecyl sulfate (SDS), a known denaturant of the enzyme. The assay was based on digestion of the cell wall of Micrococcus luteus in a suspension, a substrate sensitive to active lysozyme. Enzymatic activity against M. luteus was used to assess activity of lysozyme. The decrease in the turbidity of the cell wall suspension, a measure of relative enzyme activity, was determined by following the decrease in absorbance (at 450nm) over time using a spectrophotometer. Statistically significant greater stabilization of lysozyme was observed with Biotrue multi-purpose solution and renu fresh than with OPTI-FREE RepleniSH, OPTI-FREE EXPRESS, AQuify, COMPLETE Multi-Purpose Solution Easy Rub Formula, and a PBS control. The lysozyme activity assay revealed that Biotrue multi-purpose solution and renu fresh have the ability to stabilize lysozyme under conditions that typically denature the protein.
