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Silver nanoparticles (AgNPs) are commonly incorporated in food packaging to enhance the shelf life of fruits and vegetables. In this work, three microstructures (spherical, hematocyte-like, and pie micromorphology) of AgNPs-polylactic acid (PLA) materials were fabricated on electrospun PLA-thermoplastic polyurethane (TPU) nanofiber film by electrospraying to reduce the safety risks and the cost of food packaging materials brought by AgNPs. Among them, the spherical AgNPs-PLA microsphere/PLA-thermoplastic polyurethane (TPU) hierarchical antibacterial film had good mechanical properties, a low AgNPs content of 0.6 wt%, the highest water contact angle of 140.1°, the highest bacteriostatic rate of 99% ± 1%, and the least bacterial adhesion. The test of this composite film in strawberry packaging successfully extended the shelf life to over 7 days. The hierarchical PLA material shows promising prospects in food packaging, and the preparation method may be a potential candidate for reducing the amount of antimicrobial agent required in antibacterial materials. [Display omitted] • AgNPs/PLA microspheres are made by turning the solvent ratio of the spinning solution. • PLA-TPU films loaded with spherical AgNPs-PLA microspheres exhibit good antimicrobial and anti-adhesion properties. • The hierarchical film with low AgNPs loading can be used as food packaging materials. [ABSTRACT FROM AUTHOR]