The Brit lost the fight on a split decision, though many have questioned the outcome after a dazzling performance from the Ukrainian, who many believe should have been given a unanimous decision.
While Usyk retained the WBA, IBF and WBO titles to set up the prospect of a unification clash with Tyson Fury, he was hurt on several occasions by an underdog who was transformed from the first meeting at Tottenham Hotspur Stadium.
Joshua lacked the gas or the class to reclaim his belts, but he went out on his shield in a thriller that atoned for his passive display in London nearly a year ago.
For a second time, Usyk was operating on a different level, his movement and ringcraft keeping him away from the Briton's most dangerous shots, while his accuracy and volume of work swept him to place him in charge on the scorecards.
Joshua is no longer a dominant force in the division and another disappointment was his angry reaction after the fight when he grabbed two of Usyk's belts and dropped them out of the ring.
He also confronted the champion, asking him how he won, before then demanding the microphone to address the crowd.
"I am giving you my story. I was going to jail. I got bailed and started training my arse off, because if I got sentenced then I would not have been able to fight," Joshua said.
"I could have done better, but it showed the hard work he must have put in to beat me. Please give him a round of applause as our heavyweight champion of the world.
"I'm not a 12-round fighter. I am a new breed of heavyweight. Mike Tyson, Sonny Liston - they say 'he doesn't throw combinations like Rocky Marciano,' because I am 18 stone, I am heavy.
"It is hard work. This guy here is a phenomenal talent. We are going to cheer for him."