Barcelona suffered a first La Liga home defeat in 43 games, falling to Osasuna on the night they lost their La Liga title to Real Madrid.
Even a win over the mid-table side would not have been enough for Barca, as Madrid beat Villarreal 2-1.
Jose Arnaiz put Osasuna ahead before Lionel Messi levelled with a free-kick.
The visitors had Enric Gallego sent off for elbowing Clement Lenglet before Roberto Torres' close-range winner from Kike Barja's cross in injury time.
The last time Barca lost at home in the league was in November 2018, 4-3 to Real Betis, who were managed by now Barca boss Quique Setien.
Barca's only hope of a major trophy now is the Champions League. They host Napoli in the last 16 second leg on Saturday, 8 August, having drawn 1-1 in Italy.
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