Shale oil development in Ordos Basin, China, primarily relies on the displacement of crude oil during the post-fracturing shut-in stage (PFSIS) of horizontal wells. Reservoir wettability significantly influences the shut-in duration and even the development approach. However, due to strong heterogeneity and super tight characteristics, the reservoir usually shows an mixed wettability, and it was usually hard to differentiate the wettability in different pore sizes. With this in mind, this study focuses on core samples from shale oil reservoir in the Longdong region of the Ordos basin to quantitatively analyze the reservoir wettability. Amott method combined with nuclear magnetic resonance is adopted in the paper to meed this end. And the optimal post-fracturing shut-in duration for Huachi and Heshui areas in the Longdong region are determined based on both wettability and field practice analysis as well as numerical simulations. Qualitative wettability evaluation reveals that the reservoir in the Longdong region is weakly oil-wet (oil-wet pores account for 58.9% and water-wet pores for 41.1%), and that larger pores are more water-wet, while smaller pores are more oil-wet. Field practice observes a noticeable two-stage decline in wellhead pressure, with pressure drop rates and water content decline rates following the order of neutral reservoir > weakly oil-wet reservoir > oil-wet reservoir during the post-fracturing shut-in stage. Numerical simulations indicate that the determination of the optimal post-fracturing shut-in duration for horizontal wells should consider reservoir properties, wettability, and injection volume. The final optimal shut-in durations for the Huachi and Heshui areas in the reservoir are determined to be 36 days and 43 days, respectively. Our study qualitatively distinguishes the wettability in different pores sizes and thus determines reasonable post-fracturing shut-in durations in different areas in Longdong region. The research has major implication for building a realistic method of wettability analysis in shale or tight oil reservoir.