The mechanical properties in the outermost region of a polymer film strongly affect various material functions. We here propose a novel and promising strategy for the two-dimensional regulation of the mechanical properties of a polymer film at the water interface based on an inkjet drawing of silica nanoparticles (SNPs) underneath it. A film of poly(2-hydroxyethyl methacrylate) (PHEMA), which exhibits excellent bioinertness properties at the water interface, was well fabricated on a substrate with a pattern of SNPs. X-ray photoelectron spectroscopy and atomic force microscopy confirmed that the surface of the PHEMA film was flat and chemically homogeneous. However, the film surface was in-plane heterogeneous in stiffness due to the presence of the underlying SNP lines. It was also noted that NIH/3T3 fibroblast cells selectively adhered and formed aggregates on the areas under which an SNP line was drawn.