Studying Candida Biofilms Across Species: Experimental Models, Structural Diversity, and Clinical Implications

<i>Candida</i> biofilms play a critical role in clinical settings, contributing to persistent and device-associated infections and conferring resistance to antifungal agents, particularly in immunocompromised or hospitalized patients. Biofilm formation varies among <i>Candida</i> species, including <i>C. albicans</i> and non-albicans species, such as <i>C. glabrata</i>, <i>C. tropicalis</i>, <i>C. parapsilosis,</i> and <i>C. auris</i>, due to species-specific transcriptional networks that regulate modes of biofilm development, extracellular matrix composition, and metabolic reprogramming. These differences influence biofilm responses to treatment and the severity of infections, which can be further complicated in polymicrobial biofilms that modulate colonization and virulence. Understanding the mechanisms driving biofilm formation and interspecies interactions is essential for developing effective therapies and requires appropriate experimental models. Available models range from simplified in vitro systems to more complex ex vivo and in vivo approaches. Static in vitro models remain widely used due to their simplicity and reproducibility, but they poorly mimic physiological conditions and require careful standardization. Ex vivo tissue models offer a balance between practicality and biological relevance, enabling the study of biofilm physiology, host-microbe interactions and immune responses. In vivo models, primarily in mice, remain the gold standard for testing antifungal therapies, while alternative systems such as <i>Galleria mellonella</i> larvae provide simpler, cost-effective approaches. Advanced in vitro platforms, including organ-on-chip systems, bridge the gap between simplified tests and physiological relevance by simulating fluid dynamics, tissue architecture, and immune complexity. This review aims to examine <i>Candida</i> biofilms across species, highlighting differences in structural diversity and clinical implications, and to provide a guide to the most widely used experimental models supporting studies on <i>Candida</i> biofilm biology for the development of new therapeutic targets or drug testing.