Wound healing is a multifaceted biological process influenced by microbial dynamics, biofilms, and the choice of antiseptics. Chronic wounds often harbour biofilms—structured microbial communities that resist antimicrobial treatments and hinder healing. Among the widely available antiseptics, Povidone-Iodine (PVP-I) demonstrates exceptional efficacy in wound sanitation due to its broad-spectrum antimicrobial activity, its ability to disrupt biofilms, low resistance-inducing effect, and minimal cytotoxicity. PVP-I acts by penetrating biofilm matrices, eradicating embedded microorganisms, and reducing microbial load, thereby accelerating healing in acute, chronic, and surgical wounds. It is versatile and is available in various formulations such as solutions, ointments, and dressings, tailored to clinical needs. Comparative studies highlight PVP-I’s superior biofilm-disrupting properties and antimicrobial efficacy over alternatives like chlorhexidine, silver-based agents, and polyhexamethylene biguanide. Practical application involves integration with complementary therapies, including debridement and negative pressure wound therapy, to optimize treatment outcomes. Proposed algorithms emphasize structured care strategies, addressing infection control, biofilm management, and wound monitoring. Future developments in antiseptic therapies may leverage nanotechnology and personalized treatments based on wound microbiology and patient characteristics. PVP-I remains a cornerstone in modern wound care due to its safety, effectiveness, and versatility, offering healthcare providers a reliable tool to manage infections, promote healing, and improve patient outcomes, particularly in biofilm-associated chronic wounds.