Matthews was ruled out for the remainder of the game.
Matthews broke his right thumb on Oct. 6 while sacking Detroit Lions quarterback Matthew Stafford. The injury, diagnosed as a Bennett's fracture, required surgery to have pins implanted to stabilize the bone. Matthews missed four games.
While no timeline has been issued for Matthews' recovery, it's possible his season could be over.
"I couldn't give you a timeline," Packers coach Mike McCarthy said after the team's 38-31 loss, "but I think based off the history of what happened the first time, I'm told it's similar to [a] repeat injury."
Matthews wore a club cast when he returned on Nov. 10 against the Philadelphia Eagles but a week later he began playing with a much smaller cast, which he was wearing against the Steelers.
As he came off the field, Matthews unbuckled his chin strap with his left hand.
Nick Perry replaced Matthews for the remainder of the first half.
The Packers also lost inside linebacker Brad Jones to an ankle injury in the first half, and his return was listed as questionable. He had been listed on the injury report with an ankle injury last week. He was replaced by Jamari Lattimore.