While the literature on well-being has largely explored workers in different industries, and much has been written about patient well-being and quality of life in recent years, little attention has focused on the well-being of healthcare professionals. This study aims to provide a relevant state-of-the-art on the healthcare sector within the context of well-being. The paper employs a systematic literature review, following the PRISMA guidelines, to find relevant studies; the analysis of results is then provided by using Herzberg’s two-factor theory to explore factors affecting the well-being of healthcare workers. The literature review revealed that few relevant studies were conducted before the year 2013; only a recent increased attention to this theme, with a particular focus on nurses, enabled this study to confirm several factors individuated in the general literature on well-being and to highlight some other factors specific to healthcare. This paper provides suggestions to build a better and healthier work environment, highlighting that managers need to focus their attention on creating conditions that facilitate intra- and inter- professional relationships, on providing healthcare workers with adequate solutions for their well-being, and on strengthening the individual employee’s identification with, and involvement in, the organization.