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Background: Antimicrobial resistance is a growing public health threat and one that requires urgent attention. Antimicrobial stewardship programs are being utilized to encourage the appropriate use of antibiotics in a clinical setting, however, less has been done to address non-prescription use. There is limited research on predictors of non-prescription antibiotic use, but healthcare system distrust could be a potential motivator. The objectives of this study are to determine if there is a relationship between non-prescription antibiotic use and healthcare system distrust in the United States and Mexico, as well as how race, ethnicity, and proximity to the United States-Mexico border affect this association. Methods: This cross-sectional survey collected responses from August 2020 to August 2021 in the United States and Mexico. Data analysis included chi-square and t-tests to determine differences in characteristics by non-prescription or prescription antibiotic use. Logistic regression models were specified to calculate odds ratios (ORs) with 95% confidence intervals (CIs) to model use of non-prescription antibiotics by the level of healthcare system distrust. Models adjusted for significant confounders were fit with interaction terms for race. ethnicity, and proximity to the border for United States participants. Results: The mean healthcare system distrust score for this sample was 28.3 on a 10-50 scale (50=highest distrust) and 48.6% had used non-prescription antibiotics. After adjusting for significant covariates, the odds of using non-prescription antibiotics was 3.51 (95% CI: 1.9, 6.5) times higher for those in the highest distrust quartile versus the lowest. A significant interaction was not detected when models were fit for race, ethnicity, or proximity to the border interaction terms. Conclusions: Our results suggest that individuals with higher levels of healthcare system distrust have higher odds of using non-prescription antibiotics compared to those w