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We aimed to determine the possible inhibitory effects of zinc oxide nanoparticles (ZnO–NPs) and polyethylene glycol (PEG)-coated ZnO–NPs (ZnO–PEG–NPs) on herpes simplex virus type 1 (HSV-1). PEGylated ZnO–NPs were synthesized by the mechanical method. Antiviral activity was assessed by 50% tissue culture infectious dose (TCID₅₀) and real-time PCR assays. To confirm the antiviral activity of ZnO–NPs on expression of HSV-1 antigens, indirect immunofluorescence assay was also conducted. 200 μg/ml ZnO–PEG–NPs could result in 2.5 log₁₀ TCID₅₀ reduction in virus titer, with inhibition rate of approximately 92% in copy number of HSV-1 genomic DNA. ZnO–PEG–NPs could be proposed as a new agent for efficient HSV-1 inhibition. Our results indicated that PEGylation is effective in reducing cytotoxicity and increasing antiviral activity …