Riccardo Calafiori was due to start for Arsenal against Wigan Athletic in the FA Cup on Sunday but was injured in the warm-up.
The Italian has struggled with injury issues this season, but was thought to be ready to play in the fourth round at the Emirates on Sunday afternoon.
Calafiori was due to start at left-back, with Myles Lewis-Skelly also in the starting XI and sent out to play in midfield.
However, shortly before the start of the match there was a change to the Arsenal team, with Calafiori axed and Bukayo Saka brought into the starting XI.
Lewis-Skelly reverted to left-back, where he has played most regularly in the first, team, while Saka appeared to move into a free role in midfield.
Ceadach O’Neill, the 17-year-old Northern Irish forward, was a late addition to the bench to replace Saka.
Calafiori started the season well for Arsenal, scoring against Manchester United on the opening day of the season and remaining a regular in the starting line-up.
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However, he picked up an injury in December and a month during the busy festive period, before returning in late January.
That issue also happened in the warm-up, when he was due to play against Brighton in the Premier League but was pulled out and did not return until he played against Kairat in the Champions League on January 28.
When Calafiori began to struggle in December, Mikel Arteta said of the problem: ‘Something with Ricci was very, very strange.
‘It’s something that he felt yesterday, something very awkward that happened and today in the warm-up he wasn’t comfortable.’
He started for the first time since December in the 3-0 win over Sunderland in the Premier League on February 7.
The nature of the injury that has kept him out of the action against Wigan on Sunday is yet to be disclosed.
Already sidelined for the Gunners are Mikel Merino, Kai Havertz and Max Dowman.
Centre-back William Saliba was out due to illness but has recovered and started the game on Sunday against Wigan.