Objective: There is an increasing demand for wound healing products of natural origin. Our objective is to develop a spray formulation from Jatropha curcas L. latex extracts for wound healing applications. Materials and Methods: J. curcas L. latex was subjected to solvent extraction. The phytochemical structure was elucidated by 1 H-NMR and confirmed by LC-MS spectrometry. A topical spray formulation prepared from J. curcas latex extracts were evaluated in antimicrobial activity and radical scavenging activity. The toxicity of formulation on fibroblast cell line, collagen production and wound healing activities were carried out. Results: The 1 H-NMR and mass spectrometric analyses revealed pure compound as curcacycline A. The J. curcas latex extract formulation had radical scavenging activity and antibacterial activities. Moreover, the formulation was not toxic to the human fibroblast cells while it stimulated collagen production and healed cell injury in 24 h. Conclusion: The J. curcas latex extract promoted the wound healing after cell injury. Our findings could rise the possibility of utilizing the J. curcas latex extract spray formulation as a potential the antibacterial, antioxidant and wound healing product from nature.