: This study aims to determine the effectiveness of a new polyherbal formulation (Onopolis spray gel) which contains Centella asiatica, Aloe vera, Stichopus variegatus, and propolis extracts in HPMC and Carbopol gel bases used in the healing of second-degree burns. This study used a true experimental design with only a post-test control group. The sample used in this study consisted of 15 rats divided into 3 groups including the control, commercial control, and the Onopolis spray gel groups using a second-degree burn model made by attaching hot metal (60°C) for 5 seconds to the backs of shaved rats. The treatment lasted 14 days, with skin samples obtained on days 3, 7, and 14. Additionally, hematoxylin and eosin staining were used in making the preparations. It was also shown that the polyherbal group (Onopolis) and commercial control (Phocagel) was seen to have performed better than the control group (without medication) with re-epithelialization rates of 41.45 %, 42.85 %, and 33.19 % on the 7th day, and 70.09 %, 71.08 %, and 64.66 % on the 14th day, respectively. Conclusion: The polyherbal spray gel formula had the potential to treat second-degree burns..