A series of mesoporous NiO catalysts with high specific surface area were prepared by a simple hydrothermal method and modified by cetyltrimethylammonium bromide (CTAB) as the crystal structure directing regent. The characterization with SEM, XRD, BET, and H2-TPR results demonstrated that the introduction of CTAB effectively improved the dispersion, specific surface area, and pore volume and redox ability of NiO, and thus exposed more active sites. Meanwhile, the NiO catalyst with a CTAB/NiSO4·6H2O molar ratio of 2/3 exhibited the better catalytic ozonation performance of toluene, formaldehyde, methanol, and ethyl acetate than NiO. The in-situ DRIFTS elucidated the reaction path of catalytic ozonation of toluene and indicated that the introduction of CTAB facilitated the complete oxidation of by-products into CO2 and H2O.