Tracheostomy care is a standard procedure that nurses perform in Critical Care Units (CCUs) to reduce complications from tracheostomy. The literature indicates a clear variety of care and practice in managing tracheostomy patients within the healthcare system. This study was conducted to assess the knowledge level of tracheostomy care among nurses in CCUs in Jordanian hospitals. A cross-sectional descriptive design was used for this study. A convenience sample of 260 nurses working in the CCUs of four government hospitals completed a self-reported structured questionnaire. Data were collected from January 2021 to March 2021. A t-test and one-way Analysis of Variance (ANOVA) were used to assess the differences among socio-demographic variables in terms of knowledge score. The result revealed that the level of knowledge was suboptimal. There was a statistically significant difference in the mean level of knowledge regarding tracheostomy care (in all dimensions) by age (F = 22.595, p < 0.001), educational level (F = 355.30, p < 0.001), and work experience (F = 13.63, p < 0.001). For gender, there was a statistically significant difference in knowledge of the tracheostomy suctioning dimension (p = 0.001). The level of knowledge among nurses regarding tracheostomy care was moderate, indicating an urgent need for education.