Mario Balotelli is not the only one expected to miss the clash, with a number of first-team stars sidelined against an almost full-strength Barca.
The sides faced each other in the round of 16 last season, with Barcelona overturning a 2-0 first-leg deficit to win 4-0 at Camp Nou.
That win for Milan in the opening fixture was its first over the Liga giant since October 2004 - a run of seven fixtures.
Both clubs enjoy a storied past in the Champions League, meeting in the 1994 final when Johan Cruyff's 'Dream Team' was comprehensively dismantled by Fabio Capello's outfit 4-0.
Mario Balotelli is expected to miss the clash due to a thigh injury, while Marco Amelia will make his first appearance since April in goal, with Christian Abbiati (calf) injured and Gabriel, who played in Saturday's 1-0 win over Udinese, unregistered.
Daniele Bonera, Mattia De Sciglio, Giampaolo Pazzini (all knee) and Stephan El Shaarawy (foot) are also set to miss the clash as Milan's injury woes persist.
Barcelona failed to win for the first time under its new coach on Saturday as the club drew 0-0 with Osasuna, the first time in 64 Liga fixtures the team has drawn a blank.
Gerardo Martino has an almost full squad to choose from, with only Jordi Alba and Ibrahim Afellay (both thigh) unavailable due to injury.
Carles Puyol is likely to continue at center back, having made his first appearance since last season's 4-0 win over Milan at Osasuna on Saturday.
One man particularly keen to have a good game will be German referee Felix Brych, who will enter the contest having made headlines for all the wrong reasons in the Bundesliga on Friday as he awarded Bayer Leverkusen striker Stefan Kiessling a goal, despite the ball clearly passing through a hole in the side-netting on its way in.