The Coronavirus-19 (COVID-19) pandemic has many psychological consequences for the population, ranging from anxious-depressive symptoms and insomnia to complex post-traumatic syndromes. This study aimed to evaluate the impact of the Covid-19 pandemic on the mental well-being of healthcare workers, focusing on the association between hopelessness, death anxiety, and post-traumatic symptomatology. Eight hundred forty-two healthcare workers were recruited between 21 March 2020 and 15 May 2020. A specific questionnaire was administered to assess socio-demographic and clinical characteristics, together with psychometric scales: Beck Hopelessness Scale, Death Anxiety Scale (DAS), and Davidson Trauma Scale (DTS). Respondents with hopelessness scored higher in the DAS and DTS than respondents without hopelessness. Furthermore, death anxiety was identified as a potential mediator of the significant association between hopelessness and post-traumatic symptomatology. The impact of death anxiety should be recognized in vulnerable populations, such as frontline healthcare workers. Therefore, pharmacological and non-pharmacological strategies could be useful to attenuate the negative psychological consequences and reduce the burden worldwide.