The cytotoxicity and anti-influenza virus (IFV) activity of calcium or zinc alginate fibers were investigated to explore the feasibility of them to be used as biomaterials. African Green Monkey kidney cell (Vero) and human cervical cancer cell (Hela) cultured with alginate fibres were used to screen cytotoxic effects. After 48 h, MTT (3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H tetrazolium bromide) assays were performed. Then cytotoxicity was evaluated with six grades according to cell relative growth rate (RGR). In anti-IFV activity assay, IFV were added to all fibers and the Vero cell survival were detected by MTT assays with calculating the percentage of protection. The cytotoxity of calcium alginate fibers on Vero were grade 0 or 1 in contrast to zinc alginate fibers which was grade 0. The cytotoxity of calcium or zinc alginate fibers on Hela were grade 0. Furthermore, partial calcium or zinc alginate fibers could promote Vero or Hela cell growth. In antiviral assay the highest percentage of protection of calcium alginate fibers was 34.42%, while that of zinc alginate fibers was 59.42%. The results showed that calcium or zinc alginate fibers had a good cellular biocompatibility and the large weight zinc alginate fibers had a better anti-IFV activity than calcium alginate fibers, which is potential for tissue engineering.