Inter Miami coach Gerardo Martino announced on Monday that Argentine superstar Lionel Messi will not be available for the defending champions' Leagues Cup knockout match against reigning MLS champion Columbus on Tuesday.
Messi remains sidelined after suffering a right ankle injury a month ago in Argentina's 1-0 victory over Colombia in the Copa America final.
Messi has not yet trained with his Miami teammates since escaping a walking boot.
The Leagues Cup is a tournament with MLS and Liga MX Mexican clubs, with Messi helping lift Miami to last year's inaugural title.
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The showdown match between the Columbus Crew, last year's MLS kings, and Miami, this year's top overall team in the league table, is among eight matches on Monday and Tuesday that will decide quarter-final pairings on Friday and Saturday.
The round of 16 opens Monday with the Seattle Sounders playing host to Mexico's Pumas. Seattle defeated the Pumas in the 2022 CONCACAF Champions League final.
Other knockout matches on Tuesday include Philadelphia at Cincinnati, Cruz Azul against Mazatlan, Tigres against New York City, Toluca at Colorado, Club America at St. Louis and San Jose at Los Angeles FC.