In Italy, cultural and professional nursing improvements are reached thanks to the university-based education which marks the clinical competency and the professional autonomy in nursing decision-making. To highlight how Italian nurses perceived their nursing autonomy level in the main action-points highlighted in the Italian regulation law according to sex, age, work experience, education, shift and ward. A cohort explorative study was carried out from September 2022 to January 2023 to highlight how Italian nurses perceived their nursing autonomy levels in the main action-points highlighted in the Italian regulation law for the nursing profession according to demographic characteristics, like: gender, age, work of experience, education, shift, ward employment. A total of 403 nurses were enrolled. Significant differences were recorded in: decision-making authority in patient care and shift (p≤0.001) and ward employment (p = 0.045); ability to initiate nursing interventions and education (p < 0.001) and ward employment (p = 0.011); collaboration and communication with healthcare team members and education (p < 0.001) and ward employment (p = 0.010); independence in clinical judgment and critical thinking and shift (p < 0.001); responsibility for the planning and evaluation of nursing care and education (p = 0.005) and shift (p = 0.002) and ward employment (p = 0.013); autonomy in professional development and continuing education and shift (p < 0.001) and ward employment (p < 0.001). The results highlighted the intricate world both of the healthcare surrounding and the abilities to act autonomously within the multiprofessional staff. Future studies will develop qualitative and phenomenological designs in order to better define in which fields nurses will act their professional autonomy.