FULL PAPER Preparation, characterization, and assessment of cytotoxicity, antioxidant, antibacterial, antifungal, and cutaneous wound healing properties of titanium nanoparticles using aqueous extract of Ziziphora clinopodioides Lam leaves Niloofar Seydi 1 | Behnam Mahdavi 2 | Sogand Paydarfard 2 | Akram Zangeneh 1,3 | Mohammad Mahdi Zangeneh 1,3 | Fariba Najafi 4 | Ali R. Jalalvand 5 | Elham Pirabbasi 6 1 Department of Clinical Sciences, Faculty of Veterinary Medicine, Razi University, Kermanshah, Iran 2 Department of Chemistry, Hakim Sabzevari University, Sabzevar, Iran 3 Biotechnology and Medicinal Plants Research Center, Ilam University of Medical Sciences, Ilam, Iran 4 Department of Dermatology, School of Medicine, Kermanshah University of Medical Sciences, Kermanshah, Iran 5 Research Center of Oils and Fats, Kermanshah University of Medical Sciences, Kermanshah, Iran 6 Shoushtar Faculty of Medical Sciences, Shoushtar, Iran Correspondence Behnam Mahdavi, Department of Chemistry, Hakim Sabzevari University. Sabzevar, Iran. Email: b.mahdavi@hsu.ac.ir Mohammad Mahdi Zangeneh, Department of Clinical Sciences, Faculty of Veterinary Medicine, Razi University, Kermanshah. Iran. Email: m.mehdizangeneh@yahoo.com In recent decades, nanotechnology is growing rapidly owing to its widespread application in science and industry. The aim of the experiment was chemical characterization and evaluation of cytotoxicity, antioxidant, antibacterial, anti- fungal, and cutaneous wound healing activities of titanium nanoparticles using aqueous extract of Ziziphora clinopodioides Lam leaves (TiNPs@Ziziphora). These nanoparticles were characterized by fourier transformed infrared spec- troscopy (FT‐IR), field emission scanning electron microscopy (FE‐SEM), energy dispersive X‐ray spectroscopy (EDS), and UV–visible spectroscopy. The synthesized TiNPs@Ziziphora had great cell viability dose‐dependently (Investigating the effect of the plant on human umbilical vein endothelial cells (HUVECs) cell line) and revealed this method was nontoxic. Then, 2,2‐ diphenyl‐1‐picrylhydrazyl (DPPH) free radical scavenging test was done to assess the antioxidant properties, which indicated similar antioxidant poten- tials for TiNPs@Ziziphora and butylated hydroxytoluene. Agar diffusion tests were applied to determine the antibacterial and antifungal characteristics. Min- imum Inhibitory Concentration (MIC), Minimum Bactericidal Concentration (MBC), and Minimum Fungicidal Concentration (MFC) were specified by macro‐broth dilution assay. The data were analyzed by SPSS 21 software (Dun- can post‐hoc test). TiNPs@Ziziphora indicated higher antibacterial and anti- fungal effects than all standard antibiotics (p ≤ 0.01). Also, TiNPs@Ziziphora inhibited the growth of all bacteria at 2‐16 mg/ml concentrations and removed them at 2‐32 mg/ml concentrations (p ≤ 0.01). In case of antifungal properties of TiNPs@Ziziphora, they prevented the growth of all fungi at 2‐8 mg/ml con- centrations and destroyed them at 2‐16 mg/ml concentrations (p ≤ 0.01). In vivo experiment, after creating the cutaneous wound, the rats were randomly divided into six groups: untreated control, treatment with Eucerin basal oint- ment, treatment with 3% tetracycline ointment, treatment with 0.2% TiO 2 oint- ment, treatment with 0.2% Z. clinopodioides ointment, and treatment with 0.2% Received: 14 March 2019 Revised: 30 April 2019 Accepted: 3 May 2019 DOI: 10.1002/aoc.5009 Appl Organometal Chem. 2019;e5009. https://doi.org/10.1002/aoc.5009 © 2019 John Wiley & Sons, Ltd. wileyonlinelibrary.com/journal/aoc 1 of 14