Scans have shown that Atletico Madrid defensive duo Santiago Arias and Stefan Savic have sustained thigh injuries.
The double injury blow comes after Atletico surrendered a 2-0 first-leg lead to exit the Champions League at the hands of a Cristiano Ronaldo-inspired Juventus on Tuesday.
Atletico will therefore not be playing in the final at their Wanda Metropolitano home, but Diego Simeone's side remain in LaLiga's title race.
The injuries to Savic and Arias, confirmed in a club statement on Wednesday, could therefore be a setback for Atletico, who are already without World Cup winner Lucas Hernandez.
Atletico, seven points behind LaLiga leaders Barcelona, return to action away to Athletic Bilbao on Saturday.