Relevant personal protective measures during the COVID-19 pandemic include face masks, possibly decreasing the risk of infection among the general population and healthcare workers (HCW) if utilized properly. The aim of the study was to assess whether different Polish HCW utilize face masks according to the 2020 World Health Organization guidance (WHO) criteria. This cross-sectional study included 1156 respondents who participated in an internet survey evaluating mask-related behaviors. All the WHO criteria were complied with by 1.4% of participants, regardless of medical profession, specialty or place of employment. HCW mostly adhered to criterion 1 (C1; strict covering of the face and mouth with the mask; 90.8%), C4 (washing/disinfecting the hands after touching/taking off the mask; 49%) and C3 (taking off the mask properly without touching the anterior surface; 43.4%), whereas C2 (avoidance of touching the mask with hands) was complied with least commonly (6.8%). HCW with mask-induced itch (31.6%) complied to C2 less often (odds ratio 0.53; p = 0.01). The study reveals that Polish HCW rarely adhere to all the 2020 WHO guidance criteria on the use of masks, whereas the adherence to particular criteria is variable and may be associated with the presence of skin related conditions and other factors. Better compliance with the recommendations in the future is necessary to increase personal safety of HCW and prevent the transmission of SARS-CoV-2.