This dissertation explores the role of the wound microbiome in acute open wounds, structured into three distinct studies. The first study presents a comprehensive scoping review of the literature on Deep Sternal Wound Infection (DSWI), focusing on dimensions of the wound microbiome: microbial load, microbial diversity, presence/abundance of common wound microorganisms, and biofilm formation. Findings suggest a significant reliance on culture techniques and the need for advanced molecular methods to better understand the wound microbiome associated with DSWI. The second study investigated opioid tolerance in acute open wounds, comparing patient and wound factors, wound microbiome, and inflammatory mediators between opioid-tolerant and non-tolerant patients. Results indicate significant predictors, including younger age, higher levels of depression and resting wound pain, and reduced bacterial richness, underscoring the necessity for targeted interventions to manage pain and reduce opioid dependency in patients with acute open wounds. The third study examines the microbiome and inflammatory mediator expression in abdominal surgical wounds, differentiating between infected and non-infected cases. It identifies a higher abundance of anaerobes in infected wounds and links specific inflammatory mediators like ARG1 and MMP-8 to abdominal surgical site infections. This dissertation highlights the importance of using advanced molecular techniques and suggests future research directions to develop interventions that can modify the impact of the microbiome on wound pathogenesis and healing, potentially reducing the incidence and severity of both SSIs and opioid tolerance.