This study investigated the therapeutic effects of connective tissue manipulation (CTM) in diabetic foot ulcer (DFU). A total of 20 participants (10 in CTM group and 10 in conventional treatment group (CG)) with DFU underwent the conventional DFU treatment. In addition, the CTM group received CTM twice per week for 6 weeks. The percentage wound area reduction (PWAR) and bacterial colonization count (BCC) in log10 colony-forming units (CFU) per ml wound fluid was evaluated at baseline and six weeks. Results showed a significant change in PWAR in CTM (p L = 0.98 U = 3.81) and CG (p L = 0.26 U = 1.98). Mean reduction of BCC showed a significant reduction (p