## 1 Introduction  
Biofilms, organized microbial communities enveloped in a self-synthesized extracellular polymeric substance (EPS) matrix, significantly contribute to healthcare-associated infections (HAIs) and other microbial illnesses. EPS is a complex matrix synthesized by bacteria in biofilms, providing structural support and protection. EPS, consisting of polysaccharides, proteins, extracellular DNA (eDNA), and lipids, improves adhesion, sequesters nutrients, and protects bacteria from external pressures, such as antibiotics and immunological reactions. Its involvement in antibiotic resistance makes it a critical target for methods designed to break biofilms and address related illnesses [1; 2; 3]. The World Health Organization (WHO) predicts that in acute-care hospitals, 7% of patients in high-income countries and 15% in low- and middle-income countries would acquire at least one HAI during their hospital stay. Disturbingly, one in ten patients impacted by HAI do not survive, highlighting the urgent need for improved infection prevention and control measures [4]. In the United States, HAIs account for over 88,000 fatalities annually, with an estimated economic burden of USD 4.5 billion [5]. The yearly financial burden of HAIs in the United Kingdom is estimated at GBP 774 million, highlighting their significant impact on the healthcare system [6]. Biofilms are associated with around 65% of human microbial infections and 80% of chronic illnesses, highlighting their significant influence on healthcare systems [7; 8; 9]. Biofilm's distinctive structure and adaptive mechanisms offer defense against antimicrobial agents, positioning them as significant contributors to antimicrobial resistance. Critical elements like bacterial cell density, quorum-sensing communication, and genetic adaptations, such as efflux pump activation and mutations, contribute to their resilience, making treatment efforts more challenging [10]. The relationship between biofilms and HAIs raises significant concerns. Biofilm-forming bacteria, including _Staphylococcus aureus_, _Cagulase-negative staphylococci_ (CoNS), _Pseudomonas aeruginosa_, and _Acinetobacter baumannii_, exhibit resistance to antibiotics that can be as much as 1000 times greater than that of their planktonic counterparts [11; 12; 13]. This increased resistance extends the duration of infections and markedly elevates the likelihood of recurrence, especially in clinical settings. In demanding settings like hospitals, microbes' capacity to develop biofilms serves as a vital survival mechanism. Biofilms enable bacteria to withstand disinfectants, scarce nutrients, and antimicrobial agents, facilitating their prolonged survival on surfaces and devices [14; 15]. Furthermore, their capacity to endure toxins and adjust to ecological disturbances amplifies their influence on healthcare systems, establishing them as a significant obstacle to efficient infection control [16; 17]. To tackle the challenges presented by biofilms, it is essential to gain a more profound insight into their physicochemical and biological characteristics, as illustrated in [Figure 1]. The insights gained are crucial for formulating novel therapeutic approaches to disrupt biofilms, improve antibiotic effectiveness, and reduce their effects on healthcare. Biofilms have long been recognized as a major contributor to HAIs, posing significant challenges due to their inherent resistance to antimicrobial treatments. Previous review articles have provided detailed insights into biofilm formation and resistance mechanisms. While these reviews have significantly advanced understanding of biofilm biology, they often focus predominantly on either the mechanistic underpinnings of biofilms, creating a gap between foundational science and translational applications. Furthermore, existing reviews have not fully addressed the economic burden or operational challenges of managing biofilm-associated infections nor provided a holistic analysis of emerging therapeutic and diagnostic advancements.