Data related to the characterization of polysaccharide-based multilayer films and their interactions with tumor cells. Atomic force microscopy (AFM), contact angle measurements, and polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS) were employed to assess film morphology, surface properties, and chemical structure. In addition, images from adhesion assays with prostate tumor cells (PC-3) are provided to measure cell-film interactions.