During the ongoing multi-country monkeypox (Mpox) outbreak, healthcare workers (HCWs) have represented a key group in mitigating disease spread. The current study aimed to evaluate the attitude of nurses and physicians in Jordan towards Mpox vaccination, as well as their attitude towards compulsory vaccination against coronavirus disease 2019 (COVID-19), influenza, and Mpox. An online survey was distributed in January 2023 based on the previously validated 5C scale for psychological determinants of vaccination. Previous vaccination behavior was assessed by inquiring about the history of getting the primary and booster COVID-19 vaccination, influenza vaccine uptake during COVID-19, and any history of influenza vaccine uptake. The study sample consisted of 495 respondents: nurses (n = 302, 61.0%) and physicians (n = 193, 39.0%). Four hundred and thirty respondents (86.9%) had heard of Mpox before the study, and formed the final sample considered for Mpox knowledge analysis. Deficiencies in Mpox knowledge were reflected in a mean knowledge score of 13.3 ± 2.7 (out of 20.0 as the maximum score), with significantly lower knowledge among nurses and females. The intention to receive Mpox vaccination was reported by 28.9% of the participants (n = 143), while 33.3% were hesitant (n = 165), and 37.8% were resistant (n = 187). In multivariate analysis, Mpox vaccine acceptance was significantly associated with previous vaccination behavior, reflected in higher vaccine uptake and with higher 5C scores, while Mpox knowledge was not correlated with Mpox vaccination intention. The overall attitude towards compulsory vaccination was neutral, while a favorable attitude towards compulsory vaccination was associated with higher 5C scores and a history of previous vaccination uptake. The current study showed a low intention to get Mpox vaccination in a sample of nurses and physicians practicing in Jordan. The psychological factors and previous vaccination behavior appeared as the most significant determinants of Mpox vaccine acceptance and of attitudes towards compulsory vaccination. The consideration of these factors is central to policies and strategies aiming to promote vaccination among health professionals in efforts to prepare for future infectious disease epidemics.