A former defensive midfielder, Di Biagio last played for Ascoli Calcio 1898 in 2007, and previously also played for several other Italian clubs throughout his career, including Roma and Internazionale, in particular. At international level, he also played 31 times for the Italian national side between 1998 and 2002, scoring 2 goals, representing his country at the 1998 and 2002 World Cups, as well as at Euro 2000, where Italy reached the final.