The work aims to study effects of sputtering techniques and substrate materials on crystal structure, chemical valence and morphology of titanium oxide films. Titanium oxide films were deposited on 316L stainless steel and silicon (100) wafers) in the method of high power pulsed magnetron sputtering (HPPMS) and DC magnetron sputtering (DCMS). Plasma characteristics were tested by using optical emission spectroscopy. Crystal structure, chemical valence, grain size and surface roughness of the films were evaluated by using X-ray diffractometer (XRD), X-ray photoelectron spectroscopy (XPS), atomic force microscope (AFM) and scanning electron microscopy (SEM), respectively. The plasma characteristics showed that HPPMS exhibited higher ionization rate and plasma density than DCMS in depositing titanium oxide films. XRD results showed that titanium oxide film of pure rutile structure was successfully prepared on 316L stainless steel in the methods of HPPMS and DCMS, while both rutile and anatase structures were obtained for the films on silicone substrate. Rutile content was higher on titanium oxide film prepared in the method of HPPMS than that on titanium oxide film prepared in the method of DCMS. XPS results exhibited that higher ratio of Ti3+/Ti4+ was obtained for films deposited in the method of either HPPMS or DCMS on stainless steel compared to that on silicon substrate. SEM and hardness test results showed that titanium oxide films deposited in the method of HPPMS exhibited equiaxed grains in smaller size and of higher hardness while the films deposited in the method of DCMS exhibited a columnar crystal structure in larger size. The AFM results showed no significant difference in surface roughness of the films deposited in the method of HPPMS and DCMS. Difference in ion bombardment energy of titanium oxide samples is caused by different sputtering techniques and substrate materials, and therefore crystal structure, chemical valence and grain size of titanium oxide films are affected as well.