Bacterial biofilm-associated infections have emerged as a formidable challenge in global healthcare, posing significant hurdles to clinical treatment. Clinical data indicate that approximately 80% of bacterial infections are linked to biofilm formation, which is closely associated with the development of persistent or chronic infections. A biofilm is not a simple aggregation of bacteria but a structured community encapsulated by Extracellular Matrix (ECM) secreted by the bacteria themselves. Through multiple sophisticated mechanisms, biofilms exhibit remarkable tolerance and drug resistance, thereby severely compromising the therapeutic efficacy of conventional antibiotics. In-depth elucidation of these mechanisms and exploration of innovative therapeutic strategies are crucial for breaking the bottleneck in clinical treatment of biofilm-associated infections.