Abstract Diabetic wounds are injuries that occur in patients with diabetes that involve disturbances in the peripheral and autonomic nerves. Diabetic wounds are a major complication of diabetes and a global public health problem. There is no wound-cleansing intervention using Polyhexamethylene Biguanide as a wound-cleansing fluid against diabetic wound biofilm. To identify the effect of Polyhexamethylene Biguanide on biofilm among patients with diabetic wounds. This research was a quasi-experiment with a pre-test and post-test approach. In this study, the research subject will be given to see a pre-test before the intervention. After that, a post-test will be given to see the final results of the intervention. The sampling technique was carried out by total sampling with a total of 30 respondents. This research instrument uses a biofilm observation sheet. The statistical test used was univariate and bivariate using paired sample t-test. The results showed that the average pre-test biofilm observation score was 7,63 ± 0,490 and the post-test 3,97 ± 0,765. The results of the study showed that there was a difference in the pre-test and post-test biofilm observation scores with a p-value of 0,000. After using Polyhexamethylene Biguanide on Biofilm on Diabetic Wounds, a p-value of <0,000 (p<0,05) was obtained so that Polyhexamethylene Biguanide was effective against Biofilm wound diabetic. Patients with Diabetic wounds are expected to be able to use Polyhexamethylene Biguanide as an effort to remove Biofilm and efforts in wound healing.