This study aimed to assess the status of physical activity and its association with morphological-functional characteristics among employees of Tan Hung General Hospital, Ho Chi Minh City. The results showed that approximately 30–35% of staff did not meet recommended physical activity levels, with mean weekly MET values below World Health Organization standards. Regarding morphological-functional characteristics, mean BMI was within the overweight range, waist circumference risk was higher among physically inactive staff, visceral fat measured by DEXA was increased, cardiorespiratory fitness assessed by the IPN test was below reference levels, and biological age exceeded chronological age in the inactive group. Factors associated with insufficient physical activity included older age, female sex, lower education level, smoking, prolonged sedentary time, high blood pressure, and abnormal lipid profile. These findings indicate that insufficient physical activity remains common among healthcare workers and is closely associated with multiple health indicators, highlighting the need for workplace physical activity promotion programs in the healthcare sector.