Psychological safety is essential for rest, recovery, and fostering social connections, particularly for health and social care workers (HSCWs) who frequently operate in high-pressure environments. These workers are prone to traumatic stress, which can elevate their sense of threat and undermine their psychological safety. This study aimed to validate the Neuroception of Psychological Safety Scale (NPSS) among HSCWs in the UK (n = 443). The NPSS is based on polyvagal theory and assesses the dimensions of compassion, social engagement and bodily sensations. Internal consistency, test–retest reliability, convergent, discriminant, and concurrent validity were examined, along with the scale’s dimensionality. A three-factor structure was confirmed, with internal consistencies ranging from acceptable to excellent across subscales. Validity was supported by significant associations with measures of team psychological safety, well-being, post-traumatic stress, burnout, body perception, and personality. The NPSS also demonstrated strong test–retest reliability. These results validate the NPSS as a reliable and multidimensional tool for assessing psychological safety in health and social care settings. The study highlights the importance of psychological safety for HSCWs and provides a valuable measure to support interventions aimed at fostering safer and more supportive work environments.