This study was conducted to investigate the repair effect and mechanism of new platelet lysate film-forming spray (NPLFS) on rat skin burn wound model with a commonly used clinical dressing as the control. Forty-five SD rats were randomly divided into groups, including NPLFS treated (group A, n = 15), platelet-rich plasma (PRP) plaster group (group B, n = 15), and saline gauze cover group (group C, n = 15), fixed with 3M Tegaderm transparent film dressing. The healing rates on the 5th, 10th, 15th and 20th day, the histological hematoxylin and eosin (HE) staining changes on the 10th and 20th day, and the expression of CD31 on the 7th and 21th day after injury were observed. On the day 15 of treatment, the wounds in groups A and B were significantly reduced, while the degree of reduction in group C was not significant. On the day 20 of treatment, the wounds in groups A and B were completely healed and appeared flat, while the wounds in group C were only partly healed. On day 10, the HE staining showed significant increase in neovascularization in groups A and B as compared to group C (n = 3). The CD31 expression on day 20 after treatment showed that the integrated option densities (IODs) for rats in group A (20.13 ± 1.50) and group B (21.15 ± 0.79) were significantly higher than in group C (15.39 ± 1.41, P < 0.01). Therefore, NPLFS can promote the healing of burn wounds, and the effect is similar to that of PRP. [ABSTRACT FROM AUTHOR]