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Background: Globally, preventable medication-related problems are increasing constantly. Patient-centered communication (PCC) is essential to identify and reduce the occurrence of drug-related problems such as inappropriate use of medications, adverse drug reactions and non-adherence. In Eritrea, personal observations and anecdotal reports showed that pharmacy professionals' practice towards PCC was unsatisfactory. This study was conducted to assess pharmacy professionals' knowledge, attitude, self-efficacy and barriers towards the practice of PCC. Methods: An analytical cross-sectional study design was employed among all pharmacy professionals who were employees of the drug retail outlets and hospital out-patient pharmacies of Asmara, Eritrea. A census approach was employed and data were collected from May to June 2021 using a face to face interview-based data collection tool. Descriptive and analytical statistics including independent samples t-test and multiple linear regression were employed using IBM SPSS (version-26). Results: A total of 79 pharmacy professionals with a median age of 36 (IQR: 20) years were included in the study. The mean (SD) knowledge score of PCC was found to be 11.91 (2.0) out of 17. Moreover, the mean (SD) attitude score was 39.55 (4.0) out of 60 and the overall mean (SD) self-efficacy score was 52.51 (12.0) out of 96. Self-efficacy was significantly correlated with attitude (p=0.015). Work setting (p=0.002) and training (p=0.045) were predictors for the sub-scales of self-efficacy 'exploring patient's perspectives' and 'dealing with communicative challenges', respectively. Workload/shortage of pharmacy professionals and time constraint were found to be the main barriers of the pharmacy professionals for not practicing PCC. Conclusion: Generally the pharmacy professionals' fairly encouraging knowledge and attitude score on PCC, it was not supported with their self-efficacy. This necessitates further attention from policy makers and health facility managers in creating an ideal working environment for the pharmacy professionals to practice PCC at their best. [ABSTRACT FROM AUTHOR]