The study explored the antimicrobial and antibiofilm properties of chitosan microflowers (CMF) in sponges. The main objective was to enhance the manufacture of CMF by employing varying quantities of calcium chloride (CaCl2) and tripolyphosphate (TPP). CMF was then combined with gelatin (GE) in different proportions to produce three sponge samples: CMF0@GE, CMF1@GE, and CMF2@GE. The CMF had a morphology like that of a flower and produced surfaces with a porous sponge-like structure. The antibacterial activity, as determined by the zone of inhibition (ZOI), increased with greater doses of CMF. Among the tested samples, CMF2@GE had the greatest activity against Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, and Enterococcus faecium. CMF2@GE successfully suppressed biofilm formation, decreased clotting time to an average of 212.67 s, and exhibited excellent biocompatibility by preserving over 90 % viability of human skin fibroblast cells at dosages below 100 μg/mL. The results indicate that gelatin sponges filled with CMF have considerable promise as flexible medical instruments for wound healing and infection control.