Striker Mark Gonzalez was summoned to the Chilean national team to replace the injured Mathias Fernandez.
Fernandez felt discomfort during training in San Diego (California) on Monday. Medical tests confirmed an injury to his left knee, bad news for the defending champion of the Copa America.
The injury is a serious problem for Juan Antonio Pizzi, as Fernandez was the only playmaker in midfield.
Chile fell 1-0 to Mexico on Wednesday in their last friendly before the Copa America.
La Roja debuts next Monday against Argentina in Group D.