Introduction Patient safety culture is defined as the attitudes, perceptions, and values that staffs share within an organization related to patient safety. The safety of health care is now a major global concern. It is likely that millions of people suffer disabling injuries or death directly related to medical care. Particularly in developing and transitional countries, patient harm is a global public health problem. The objective of the study is to assess patient safety culture and associated factors among health care professionals working in public hospitals in Dessie town, North East Ethiopia, 2019. Methods Facility based quantitative study was employed from March 15 -April 30, 2019 in public hospitals in Dessie town. Four hundred and twenty two health care professionals were recruited to complete a structured pretested self-administered questionnaire. The data was cleaned, coded and entered in to Epi Info-7 and exported to SPSS version 20. Data was further analyzed using bivariate and multivariate logistic regression analyses. Variables with P value of less than 0.05 in multivariate analysis were declared as statistically significant at 95% CI. Results Of the 422 recruited a total of 411 participants completed the survey with a response rate of 97.4%. Close to half (184(44.8%)) of the participants indicated good patient safety culture. Good patient safety culture was positively associated with working in primary hospital (AOR = 2.56, 95% CI = 1.56, 4.21). On the other hand, good patient safety culture was negatively associated with health professional's age between 25-34 year (AOR = 0.25, 95% CI = 0.08-0.74) and working in Pediatrics ward (AOR = 0.39, 95% CI = 0.17-0.9) and in emergency ward (AOR = O.25, 95%CI = 0.09-0.67). Conclusion The overall level of patient safety culture was under 50%. Good patient safety culture had positive association with working in primary hospital and negative association with professionals' age between 25-29 year, 30-34 year and working in pediatrics and emergency ward. Implementing actions that support all dimensions of safety culture should be promoted at all levels of hospitals.