Background: The COVID-19 pandemic underscored the effect of different vocal demands (including the highly stressful conditions to address health disparities) on the interactions between healthcare providers, patients, and the public. Challenges in healthcare voice production during the pandemic were documented, focusing on the effect of wearing surgical masks on voice analysis results. Interestingly, in the post-COVID era, the analysis of the impact of vocal demands (including working conditions) on voice production and the well-being of healthcare workers, and strategies to mitigate this relationship is limited. Methods: This cross-sectional study was conducted in 2021 at a Colombian healthcare facility. The study population comprised 79 healthcare workers, including physicians, registered nurses, assistant nurses, therapists, and administrative staff from different clinical and non-clinical areas. Working conditions and health-related data were collected through a comprehensive online survey. Results: The multivariate analysis showed that female registered nurses spending longer in transportation to work, with self-perceived inadequate background noise conditions, monotony at work, and throat clearing had statistically significant higher values of jitter, whereas working on Information management, radiology, intensive care, or therapies and self-reported high levels of stress decreased the values of this parameter. Increased HNR was associated, in the multivariate model, with marital status, female gender, being a medical doctor working on information management, radiology or therapy, phonation with the surgical mask, deviated nasal septum, sleep duration in minutes, and apnea. Conclusion: This study highlights the significant impact of vocal demands and working conditions on healthcare workers' voice production and well-being during the COVID-19 pandemic. The findings reveal that factors such as transportation time, background noise, work monotony, high stress levels, the use of surgical masks, marital status, and sleep patterns influenced voice production. These results underscore the need for targeted strategies to promote healthy voice use and improve the overall well-being of healthcare workers in the post-COVID era.