The　objective　of　this　study　was　to　design　a　drug　delivery　system　consisting　of　biotinylated　cholesterol-modified　glycol　chitosan　(Bio-CHGC)　nanoparticles　and　fish　collagen/chitosan　(Col/Ch)　film　for　localized　chemotherapy.　Bio-CHGC　was　synthesized,　and　then　its　self-assembled　nanoparticles　were　prepared　by　probe　sonication.　Doxorubicin　(DOX)-loaded　Bio-CHGC　(DBC)　nanoparticles　prepared　by　dialysis　had　spherical　shape,　and　their　sizes　were　in　the　range　of　330-397.　nm.　Col/Ch/DBC　nanoparticle　films　were　fabricated　by　freeze-drying.　SEM　showed　that　the　DBC　nanoparticles　were　uniformly　distributed　into　the　films,　and　the　films　retained　their　structural　integrity.　A　higher　degradation　and　swelling　rate　of　the　drug　films　led　to　a　higher　diffusion　rate　of　the　nanoparticles　from　the　films,　resulting　in　an　increase　in　the　drug　release　from　nanoparticles.　The　release　of　DOX　from　the　films　or　Bio-CHGC　nanoparticles　was　sensitive　to　the　pH　value　of　the　release　medium.　In　addition,　the　DOX　release　ratio　of　the　drug　films　was　lower　than　that　of　the　nanoparticles　alone,　suggesting　that　the　drug　films　had　a　double-sustained　effect　on　the　drug　release.　MTT　assay　implied　that　the　DBC　nanoparticle　film　showed　a　higher　inhibitory　ratio　than　the　film　containing　nanoparticles　without　biotin,　indicating　that　biotin　moieties　in　the　nanoparticles　played　an　important　role　in　exerting　a　cytotoxic　effect.　These　data　demonstrate　that　Col/Ch/DBC　nanoparticle　film　has　the　potential　to　be　used　as　a　localized　delivery　system　for　hydrophobic　antitumor　drugs.　©　2015　Elsevier　B.V.