The discovery of the existence of biofilms in the 1970s has resulted in a greater understanding of how bacteria form a synergistic relationship to facilitate their survival by incorporating and sharing the individual attributes that individual bacteria have with the polymicrobial flora that composes the biofilm. Advances in molecular biology have aided in delineating complexity of biofilms, but the abundance and plethora of the microbial world provides an ongoing and daunting task. The enormity of this task is reflected in the chronic non-healing wound, as it is estimated that 60% of all chronic non-healing wounds have a biofilm component. The etiology of the wound, its anatomical location, the chronicity of the wound, and the varying polymicrobial population that exists in the varying clinical settings where treatment occurs, adds to the difficulty of this problem. A review of the present existing knowledge of biofilms in the chronic non-healing wound and their clinical relevance is discussed. The relevance of clinical findings, readily available laboratory tests, and an analysis of the primary mode of action of certain treatment options are also discussed as to their effectiveness in the treatment of chronic non-healing wounds.