Objective To explore the effect of recombinant human basic fibroblast growth factor (rh-bFGF) spray combined with ultra-pulsed CO2 fractional laser in the treatment of acne scars. Methods A total of 64 patients with acne scars admitted to the Department of Dermatology of Yancheng No.1 People's Hospital from January to December 2022 were selected as the research objects. They were divided into control group and study group by random number table method, with 32 patients in each group. The control group was treated with ultra-pulsed CO2 fractional laser combined with mupirocin ointment, and the study group was treated with ultra-pulsed CO2 fractional laser combined with rh-bFGF spray. The clinical efficacy, acne scar, acne basal depth, skin barrier function and adverse reactions were compared between the two groups. Results The total effective rate of the study group was 96.88%, which was higher than 81.25% of the control group (P <0.05). After treatment, the score of acne scar scale in the study group was lower than that in the control group (P <0.05). After treatment, the depth of acne base in the study group was lower than that in the control group (P <0.05). After treatment, the TEWL, pH value and erythema index of the study group were lower than those of the control group, and the water content of the stratum corneum was higher than that of the control group (P <0.05). The incidence of adverse reactions in the study group was 9.38%, which was lower than 15.63% in the control group, but the difference was not statistically significant (P >0.05). Conclusion rh-bFGF spray combined with ultra-pulsed CO2 fractional laser is effective in the treatment of acne scars, which can improve the scar condition and repair the skin barrier without increasing the incidence of adverse reactions. It is worthy of clinical application.