In the present study, we aimed to assess the prevalence and the relationships between alexithymia, burnout and hopelessness in a large sample of healthcare workers (HCWs) during the third wave of Covid-19 in Italy. Alexithymia was evaluated by the Italian version of the 20-item Toronto Alexithymia Scale (TAS-20), hopelessness was measured using the Beck Hopelessness Scale (BHS) and irritability (IRR), depression (DEP) and anxiety (ANX) were evaluated with the Italian version of Irritability‚ Depression‚ Anxiety Scale (IDA). This cross-sectional study recruited a sample of 1445 HCWs from a large urban healthcare facility in Italy from 1 June—31 May 2021. Comparison between individuals positive (n=214, 14.8%) or not for alexithymia (n=1231, 85.2%) controlling for age, gender and working seniority revealed that positive subjects showed higher scores on BHS, MBI, IRR, DEP and ANX than not positive ones (p<0.001). In the linear regression model, higher working seniority and higher MBI, DEP, ANX and TAS-20 scores were associated with higher hopelessness. In conclusion, increased hopelessness was associated with higher burnout and alexithymia. Comprehensive strategies should be implemented to support HCWs mental health and mitigate the negative consequences of alexithymia, burnout, and hopelessness