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Christine Selhuber-Unkel, Sören Björn Gutekunst; Microstructured materials for removing Acanthamoeba castellanii from contact lens storage cases. Invest. Ophthalmol. Vis. Sci. 2016;57(12):1455.
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© ARVO (1962-2015); The Authors (2016-present)
Acanthamoeba castellanii (A. castellanii) cause acanthamoeba keratitis, which is a serious infection of the eye, and 85% of acanthamoeba keratitis cases are related to wrong contact lens usage. Many cases have also been related to the resistance of A. castellanii cysts to contact lens cleaning solutions. We hypothesize that microporous hydrogels containing microchannels in combination with larger cavities are able to capture A. castellanii.
Microporous polyacrylamide hydrogels were fabricated by incorporating polyacrylamide into interconnected sacrificial ZnO networks. The ZnO was dissolved by hydrochloric acid, leaving behind seamlessly interconnected microchannels in combination with larger cavities. Prior to biological experiments, the microchannel-containing polyacrylamide was washed for at least five days in sterile PYG 712 medium and incubated in cAMP for inducing A. castellanii chemotaxis.A. castellanii trophozoites were grown under axenic conditions to establish a cyst-free culture. They were seeded onto the polyacrylamide structure and incubated for at least 30 min. A. castellanii migration was investigated using light microscopy. Image processing tools (ImageJ) were used to analyze the migration speed of A. castellanii trophozoites as a function of microchannel diameter (number of investigated trophozoites: N=80). Control experiments were carried out on bulk polyacrylamide (N=9).
The speed of A. castellanii trophozoites inside the microchannel network was significantly reduced compared to bulk materials (v(channel) 155 µm ± 50 µm/sec; v(bulk)=255 µm ± 80 µm/sec). Furthermore, 55 % of A. castellanii trophozoites in microchannel hydrogels were resting for at least 10 min (N=35) in cavities, whereas no resting trophozoites have been observed on bulk materials (N=18). A. castellanii trophozoites stayed alive for 6 days (N=23), as verified by intracellular motion.
The obtained results are consistent with our hypothesis that microstructured hydrogels are able to attract A. castellannii, thus providing a novel strategy to remove these human pathogens with the hydrogel material from contact lens storage cases.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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