The present study demonstrated collagen nanofiber deposition onto titanium and partially stabilized zirconia, yttria-stabilized zirconia (Y-TZP), using an electrospray deposition (ESD) technique. Type I collagen was dissolved into 1,1,1,2,2,2-hexafluoro-2-propanol at different concentrations. The effects of the flow rate of the collagen solution and the applied voltage on collagen nanofiber deposition were investigated.For titanium substrates, a collagen concentration of 50 mg/ml, a flow rate of 5.0 µl/min, and an applied voltage of 16 kV were found to be suitable for the deposition of uniform collagen nanofibers. Collagen nanofibers could be sprayed onto Y-TZP, which is not an electrically conductive substrate, although nanofibers fused during ESD. To avoid the fusion of nanofibers, titanium sputter-coating was performed on Y-TZP. Less fusion of collagen fibers occurred. A collagen concentration of 50 mg/ml, a flow rate of 5.0 µl/min, and an applied voltage of 25 kV were found to be appropriate for the deposition of uniform collagen nanofibers onto titanium sputter-coated Y-TZP. The ESD technique was shown to be effective for the deposition of collagen nanofibers.