No reports have discussed this point before that. Chitosan nanospheres (CNPs) with an average size of 110 nm and positive potential of +27.8 mv have been prepared via chemical ion gelation method and characterized by spectroscopy and electron microscopy. Hep G2 cell line is a suitable host for Hepatitis C Virus (HCV) replication. Hep G2 was treated with a concentration of 100μg/mL (w/v) of CNPsto investigate their effect on Hep G2. Visible cell aggregations and granulations took place in the tissue culture flask after 48 hours. This may be mediated, in part; by Hep G2 phagocytosis, which has been recorded with electron microscopy 24 hours post treatment. Thus, CNPs might attain the critical aim of drug delivery in a persistent HCV culture