Pathogenic biofilms are considered a potential major obstacle to healing chronic wounds. The six-month evaluation period in this study assessed the effect of a single treatment of the HYBENX® (HYB) Root Canal Cleanser on chronic wounds, especially its ease of use, safety, potential for shortening duration of Inflammatory Phase, and promotion of granulation. HYB gel was applied to the wound bed and periwound skin for 10 seconds. Gel was removed by cotton gauze pads and low-pressure saline rinsing. Standard wound dressings based on wound etiology, location, and exudate characteristics were applied. Duration of these seven HYB-treated wound cases (two pressure ulcers, one surgical wound, four lower extremity ulcers-a venous reflux-associated calf ulcer, one diabetic foot ulcer, and two animal-induced wounds-brown recluse spider and bovine bite) ranged from 12 days to 10 years. Three cases had sinus tracts. After HYB application, sinus tract closure occurred at 7, 16, and 21 days. The 10-year recalcitrant wound from brown recluse spider bite healed in 97 days. Diabetic foot ulcer responded to HYB treatment plus additional strategies and healed in six months. These data support the hypotheses that pathogenic biofilm actively prevents the healing of chronic wounds, and biofilm can be disrupted with a single HYB treatment., (Copyright 2020 Biolife Sas. www.biolifesas.org.)