Background: COVID-19 is a highly communicable disease that can be transmitted from animal-to-human and human-to-human contacts. It is still now a major global threat for which vaccination remains the ultimate solution to protect it, especially healthcare professionals are the first frontiers to fight against the COVID-19 virus which makes them at higher risk of this disease. Therefore, to reduce the spread of COVID-19, we need to improve the acceptance of COVID-19 vaccines is crucial. The aim of this study was to assess the vaccine hesitancy of the COVID-19 vaccine among health professionals who worked at the University of Gondar Specialized Hospital. Methods: A cross-sectional survey was conducted between 1 May and 10 June 2021. And the data were analyzed using Statistical Package for Social Sciences version 26. A chi-square test was conducted and to assess the associations between socio-demographic characteristics, perceived risk of COVID-19, attitude, and acceptance of COVID-19 vaccine, a p-value of less than 0.05 was considered to declare statistical significance. Results: We surveyed 319 health professionals with a response rate of 67.87%. Of these, 74.9% (239) were male, and 81.2% (259) of health professionals were perceived exposed to COVID-19 without testing. 87.7% of respondents would like to vaccinate their parents. 66.2% (208) of them have accepted the COVID-19 vaccine. 54.85% (175) of health professionals had a good attitude and 45.8% (146) of health professionals were received the first-round COVID-19 vaccine. Conclusion: This study showed that more than 60% of health professionals had good acceptance of COVID-19 vaccination during the pandemic period. Regarding vaccine safety profile may reduce the vaccination in the future. To increase vaccine uptake in response to the COVID-19 pandemic, COVID-19 vaccination programs should be redesigned to remove barriers to vaccine acceptance. Knowledge and attitude toward the COVID-19 vaccine should be promoted.