Carlo Ancelotti's side will also now be sweating over the fitness of the France international for next week's Champions League semi-final second leg against Bayern Munich.
According to journalist Guillermo Rai, the midfielder's foot injury is not serious, but it is likely to rule him out of Saturday's game against Cadiz.
Tchouameni would have likely been rested against Cadiz regardless of his fitness, with Ancelotti preparing to remove a number of key players from the XI ahead of the second leg of their European Cup semi-final at Bernabeu on May 8.
The 24-year-old has been a vital player for Real Madrid this season, making 37 appearances in all competitions, scoring three times and providing one assist in the process.
Aurelien Tchouameni's foot injury 'regarded as minor'
Tchouameni's ability to operate as a centre-back has been vital for Real Madrid this season, with David Alaba out since last December, and Eder Militao only just back from a lengthy absence.
The Frenchman started in central midfield against Bayern on Tuesday, but he dropped back into the middle of the defence when Nacho was replaced in the second half of the 2-2 draw.
Providing that Tchouameni is fit for the second leg with Bayern, Ancelotti must decide whether to name the ex-Monaco youngster in central defence alongside Antonio Rudiger, or keep him in the middle of midfield.
There is set to be a huge injury boost for Real Madrid this weekend, though, with number one goalkeeper Thibaut Courtois in line to make his first appearance of the campaign.
Thibaut Courtois is expected to start against Cadiz
Courtois suffered an ACL injury in training ahead of Real Madrid's opening match of the 2023-24 La Liga campaign; he was due back at the end of March, but it was then revealed that the Belgian had suffered another knee problem, which required an operation.
The club's first-choice stopper is now back to full fitness, though, and he is set to be between the sticks for the clash with Cadiz on Saturday.
Real Madrid will be confirmed as La Liga champions with four matches to spare this weekend if they overcome Cadiz and second-placed Barcelona fail to beat third-placed Girona.
Ancelotti is set to ring the changes for the clash against Cadiz, with Fran Garcia, Arda Guler, Brahim Diaz, Dani Ceballos and Joselu among those expected to start.
Dani Carvajal will also be back in the squad after missing the game against Bayern due to a European ban.