The Gunners defender had to be substituted during the early stages of the second half in their 5-2 win at West Ham on Saturday evening.
Calafiori appeared to hold his groin once he was off the pitch in what may be a blow to Arsenal's plans for United on Wednesday night.
The versatile Italian, who was replaced by Oleksandr Zinchenko, has quickly established himself as a firm fan favourite amongst the Gunners faithful.
He only returned to the starting XI last week against Nottingham Forest having missed six games with a knee injury.
And Mikel Arteta will be left sweating over his fitness, with fellow full-backs Ben White and Takehiro Tomiyasu already out due to injury.
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There was another worry for Arsenal moments after Calafiori was hauled off before the hour-mark.
Bukayo Saka, who scored a penalty against West Ham and assisted two other goals, went down in pain on the London Stadium turf.
However, despite a long pause for treatment and a brief spell off the pitch, he was able to continue playing.
Though he was later taken off with Raheem Sterling coming on in his place in the closing stages.
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Arsenal were able to get through the rest of the match without anymore worries of their players' fitness in what was a thumping win.
Gabriel headed in the opener with only ten minutes played, doing so from yet another set-piece that was crossed in by Saka.
Leandro Trossard doubled Arsenal's lead before with less than half an hour on the clock - tapping home into an empty net after being picked out by Saka.
All three points were pretty much sealed moments later when Martin Odegaard scored from the penalty spot.
Kai Havertz continued the romp when he added the Gunners' fourth, although hosts West Ham did their best to fight back.
Aaron Wan-Bissaka's finish was followed by an excellent free-kick from Emerson Palmieri to ensure Arsenal only had a two-goal advantage.
However, the north London outfit scored their fifth and final goal shortly before half-time thanks to Saka's penalty.
Victory has sent them second in the Premier League and six points behind Liverpool, who host Manchester City tomorrow.
Arsenal could drop down in the standings on Sunday though should Chelsea beat Aston Villa by more than three goals.
Arteta's side will now turn their focus towards United's visit at the Emirates on Wednesday as they continue to hunt down Liverpool.