A series of new nonionic antimicrobial polymers with a biodegradable polyvinyl alcohol (PVA) backbone grafted with indole units and different hydrophobic alkyl or ether groups were synthesized by facile esterification. The chemical structures and thermal properties of the obtained polymers were characterized by GPC, NMR, FTIR, WAXD, TGA and DSC analyses. All these nonionic polymers showed a significant antibacterial effect similar to gentamicin against 9 food and human pathogenic bacteria according to the disk diffusion assay. The presence of alkyl or ether groups in most cases did not significantly affect the antibacterial effect compared to the polymer with unsubstituted indole units (with N–H moieties). The impacts of the OH conversion and molecular weight of the obtained polymers on their antimicrobial and anti-quorum sensing effects were also preliminarily investigated. Finally, the obtained indole-grafted PVAs were subjected to MTT assay using a mammalian cell line and hemolysis investigations, and the results showed excellent biocompatibility, particularly for those with ether substituents.