The application of nanotechnologies to medicine, or nanomedicine, which has already demonstrated its tremendous impact on the pharmaceutical and biotechnology industries, is rapidly becoming a major driving force behind ongoing developments in the antimicrobial and wound healing fields. This study confirms the potential of Allium eriophyllum Boiss aqueous extract for the green synthesis of titanium nanoparticles (TiNPs). Also, we reveal the antioxidant, cytotoxicity, cutaneous wound healing, antifungal and antibacterial properties of TiNPs. These nanoparticles were characterized using Fourier transform infrared (FT-IR) and UV-visible spectroscopies, X-ray diffraction (XRD), transmission electron microscopy (TEM) and field emission scanning electron microscopy (SEM). FT-IR findings suggested molecular interactions of compounds in the plant were the sources of reducing power, reducing titanium ions to TiNPs. In XRD analysis, 22.8 nm was measured for the crystal size of the nanoparticles. SEM and TEM images indicated a uniform spherical morphology and average diameters of 22 nm. 2,2-Diphenyl-1-picrylhydrazyl experiments were conducted to assess the antioxidant activities, which indicated similar antioxidant potentials for TiNPs and butylated hydroxytoluene. In the antimicrobial part of this study, agar diffusion experiments were done to determine the antibacterial and antifungal characteristics. TiNPs had antifungal activities against Candida guilliermondii, C. krusei, C. albicans and C. glabrata and antibacterial potentials against Gram-negative bacteria (Pseudomonas aeruginosa, Salmonella typhimurium and Escherichia coli O157:H7) and Gram-positive bacteria (Staphylococcus aureus, Streptococcus pneumoniae and Bacillus subtilis). Also, these nanoparticles did not have any cytotoxicity against human umbilical vein endothelial cells. In in vivo experiments, TiNP ointment significantly decreased (p ≤ 0.01) wound area, total cells, macrophages, neutrophils and lymphocytes and notably increased (p ≤ 0.01) wound contracture, vessels, hydroxyproline, hexosamine, hexuronic acid, fibrocytes and fibrocyte/fibroblast ratio in rats. The results of FT-IR, UV, XRD, TEM and SEM analyses confirm that aqueous extract of A. eriophyllum leaves can be used to yield TiNPs with notable antioxidant, antibacterial, antifungal and