Objective To evaluate the safety and efficacy of non-invasive ventilation with aerosolized surfactant (AS) in the treatment of premature infants with respiratory distress syndrome (RDS). Methods From July 2019 to June 2020, in this unblinded Phase I study, the premature infants with RDS who met the criteria with the informed consent of their guardians were enrolled. They were treated with nasal intermittent positive pressure ventilation (NIPPV) and received one dose (100 mg/kg) of aerosolized surfactant by vibrating mesh system (Aeroneb Solo) after hospitalization. The vital signs, adverse reactions and blood gas during areosolizing were recorded and prognosis of them also recorded in detail. Results Twenty infants were enrolled, nineteen completed the study, one died in 26 days. Five infants still required endotracheal intubation and mechanical ventilation after AS treatment. One hour after AS treatment, infants' oxygen saturation and indicators of pulse oximetry improved (P＜0.05), and heart rate decreased (P＜0.05) , but blood pressure and parameters of ventilator had no change (P＞0.05). Infants all tolerated the aerosol treatment well. No other significant adverse events were identified. Conclusion We have demonstrated the feasibility and safety of AS treatment in preterm infants with RDS receiving non-invasive respiratory support. The treatment was well tolerated by infants and clinical caregivers , but still need further study.