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Nijani Nagaarudkumaran, David Joseph McCanna, William Ngo, Lyndon William Jones; In vitro Quantification of Cytokines Adhered to Contemporary Contact Lens Materials. Invest. Ophthalmol. Vis. Sci. 2020;61(7):1490.
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© ARVO (1962-2015); The Authors (2016-present)
Contact lenses (CL) may induce a low-level inflammatory response on the ocular surface. Previous studies have quantified the concentration of inflammatory mediators present in the tear film during CL wear. Analyzing the inflammatory mediators loosely adhered to CL materials may provide another perspective on the role that contact lenses play in inflammation. The purpose of this in vitro study was to quantify a variety of cytokines found in the tear film that adhered to various CL materials and to develop a method that could extract them.
Cytokines IL-1β, IL-6, IL-8, and TNF-α (Meso Scale Diagnostics, Rockville, MD) were combined with 5 mL of Diluent 2 to prepare a cytokine solution with a final concentration of 119.41, 166.05, 101.48 and 40.73 pg/mL, respectively. Contact lenses (etafilcon A, somofilcon A, omafilcon A, delefilcon A) (n=4 each) were each placed into a polypropylene tube containing a volume of 200 μL of the prepared cytokine solution and were incubated at 23°C for 6 hours. The lenses were removed from the tubes using tweezers and placed into a 0.6 mL microcentrifuge tube containing 200 μL of Diluent 2 and were incubated at 23°C for 1 hour. The microcentrifuge tube was then vortexed for 5 seconds and pin sized holes were made at the base of the tube. The tube was then placed into a larger 2.0 mL microcentrifuge tube acting as a carrier and were centrifuged at 604 RCF. The eluent in the 2.0 mL microcentrifuge was then collected and stored at -80°C for cytokine quantification at a later date, using the MESO QuickPlex SQ 120 (Meso Scale Diagnostics, Rockville, MD). Statistical analysis was performed using a one-way ANOVA.
There was no significant difference between cytokine concentrations for all CL materials (p>0.05).
While there were no significant differences between the concentrations of cytokines found loosely adhered to the soft CL materials investigated, the results support this method as a means to quantify such cytokines on soft lens materials. This method may be used to examine human-worn lenses in future studies.
This is a 2020 ARVO Annual Meeting abstract.
Figure 1: Concentration of loosely adhered IL-1β (A), IL-6 (B), IL-8 (C), and TNF-α (D) in 200μL of eluent from each lens material.
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