In this work, the technique of pulsed laser ablation in liquid (PLAL) was used to synthesize Zirconium oxide (ZrO2) colloidal Nanoparticles in Distilled Water (D.W.). The optical properties and The morphology of the Nanoparticles were characterized by (UV-Vis) spectrophotometer, X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR) and field emission scanning electron microscopy (FE-SEM) analysis. study have been the optical properties of the (ZrO2) Nanoparticles by (UV-Vis) analysis all colloidal samples have shown broad peak at wavelength of 289 nm for Zirconium oxide Nanoparticles produced by technology pulsed laser ablation in liquid. The structural analysis by X-ray diffraction (XRD) reveals the formation of mixture of monoclinic (m-ZrO2) and tetragonal (t-ZrO2) phases of nano crystalline Zirconium oxide Nanoparticles produced by technology pulsed laser ablation in Distilled Water. The Fourier transform infrared spectroscopy (FT-IR) study determined the functional groups of (ZrO2) Nanoparticles in Distilled Water. verified the nanostructure of produced materials confirmed by field emission scanning electron microscope (FE-SEM) analysis. Biocompatibility tested For zirconium oxide solutions prepared with pulse laser ablation technology in distilled water and using three different types from bacteria which includes gram-positive (Staphylococcus aureus) bacteria And gram-negative (Pseudomonas aeruginosa) and (E.coli) bacteria as the results showed that zirconium oxide nanoparticles it has biocompatibility Due to the absence of any toxicity to the tested bacteria.