To compare the Vickers microhardness, surface roughness, initial adhesion, and osteogenic differentiation on titanium (Ti) and nitrurized titanium (NTi) plates were treated by UV irradiation and chitosan. Each plate was subjected to Vickers hardness with a pressure of 2.9 N for 10 seconds and roughness evaluation by atomic force microscope (AFM) analysis. Three groups of each type of plates were tested: control (C), ultraviolet irradiation (UV), and chitosan (Q). The UV group was exposed to UV-irradiation for 20 min at 253.7 nm (52 μW/cm2). The Q group was coated with 1% chitosan, and the C group had no treatment. The osteoblasts (2 × 106 cells/mL) were inoculated in each group for 60 min and their viability was determined by the MTT bioassay. Osteogenic differentiation was performed over 4 weeks and determined by alizarin red staining. The mean was analyzed with the Shapiro-Wilks, Kruskall-Wallis, and Mann-Whitney U tests of normality (n = 9/gp). The NTi plates hardness (125.1 ± 4.01 HV) was higher (P = 0.026) than the Ti plates (121.3 ± 2.23 HV). The surface topography was: NTi (Ra = 0.098 μm) and Ti (Ra = 0.212 μm). The quantification of cell adhesion was: Ti + Q = 123 ± 4.9% (P < 0.05) < NTi + Q = 107 ± 3.3% < Ti = 100 ± 10.7% < NTi = 72 ± 6.8% < NTi + UV = 71 ± 4.4% < Ti + UV = 69 ± 3.5%, regardless the plates, the presence of chitosan induce a faster osteogenic differentiation. The Ti + Q plates tested the highest cell attachment and osteogenic adhesion suggesting their potential use of chitosan for cell-implant interaction.