Differential effects of graphene oxide nanosheets on Candida albicans phagocytosis by murine peritoneal macrophages


Macrophages, as effector cells involved in the innate and adaptive immunity, play a key role in the response to nanomaterials as graphene oxide (GO) and in their cellular uptake. The interactions at the interface of GO nanosheets, macrophages and microbial pathogens need to be assessed to determine the possible impairment of the immune system induced by biomedical treatments with this nanomaterial. Here, we have evaluated by flow cytometry and confocal microscopy the ability of murine peritoneal macrophages to phagocytose the fungal pathogen Candida albicans, alive or heat-killed, after treatment with poly(ethylene glycol-amine)-derivatized GO nanosheets (PEG-GO). After GO treatment, differences in fungal phagocytosis were observed between macrophages that had taken up GO nanosheets (GO* population) and those that had not (GO(-) population). GO treatment increased the ingested alive yeasts in GO(-) macrophages, whereas phagocytosis diminished in the GO' population. Ingestion of heat-killed yeasts was slightly higher in both GO(-) and GO(+) populations when comparing with control macrophages. For the first time, we show that GO uptake by macrophages modulates its phagocytic capability, affecting differentially the subsequent ingestion of either alive or heat-killed yeasts. Enhanced ingestion of heat killed yeast by GO-treated macrophages suggests a beneficial role of this nanomaterial for the clearance of dead microorganisms during infection. (C) 2017 Elsevier Inc. All rights reserved.



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Diez-Orejas, R; Feito, MJ; Cicuendez, M; Rojo, JM; Portoles, MT

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This study was supported by research grants from the Spanish Ministerio de Economia y Competitividad (MAT2013-43299-R and MAT2016-75611-R AEI/FEDER, UE). M.C. acknowledges the financial support from the Portuguese FCT [SFRH/BPD/101468/2014 Post-Doctoral grant]. J.M. Rojo is supported by Grant PI13/01809 from "AES, Plan Estatal I+D+i, ISCIII-Subdireccion General de Evaluacion y Fomento de la Investigacion, Spanish Ministerio de Economia y Competitividad (MINECO). The authors wish to thank Dr. D. Prieto and Dr. J. Pla for kindly providing the RFP C albicans strain, R. Alonso for scientific discussion and L. Casarrubios for statistical analysis. Thanks also to the staff of the Centro de Citometria y Microscopia de Fluorescencia of the Universidad Complutense de Madrid (Spain) and ICTS Centro Nacional de Microscopia Electronica (Spain) for the assistance in the flow cytometry, confocal microscopy and AFM studies, respectively.

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