A systematic experimental investigation concerning the fretting-induced tribo-chemical state and its effect on the fretting wear behavior of titanium alloys under the vacuum atmospheres (4 × 10–3 Pa and 4 × 10–1 Pa) in different fretting regimes is reported. An in situ XPS analysis tester was developed to capture the real tribo-chemical state of worn surface for all test conditions. Results show that samples subjected to different vacuum atmospheres have varied tribo-chemical states depending on the fretting regime, which play significantly different roles in determining the associated damage mechanisms and the resulting fretting wear resistance. Under both vacuum atmospheres, in the partial slip regime (PSR) the worn scars were mainly covered by TiO2, showing comparable levels of very slight damage, while in the mixed fretting regime (MFR), the tribo-layer is still mainly consisted of TiO2, but with an evident peak of Ti metal for the high vacuum degree (4 × 10–3 Pa) in MFR, showing a mild damage. In contrast, in the gross slip regime (GSR), Ti metal was prone to be oxidized to Ti2O3 and TiO on the worn scar, especially for the low vacuum degree (4 × 10–1 Pa) having a highest content of Ti2O3. It might be inferred that the tribo-layer containing more Ti2O3 formed during fretting wear process is susceptible to be broken, hence showing a highest fretting wear volume in GSR for the low vacuum degree. The results suggest that for the vacuum environments, the Ti6Al4V may be more suitable to be used under the high vacuum atmosphere.