Barkley finished with 57 yards on 13 carries and had two catches for 12 yards.
Barkley said on Tuesday that playing two games four days apart while coming back from a torn ACL in his right knee was not ideal.
He tore the knee ligament in Week 2 of last season, had surgery in October and started the summer on the physically unable to perform list. He didn't face his first contact until just a few weeks ago and was feeling natural soreness after being tackled for the first time in almost a year in Week 1 on Sunday.
Barkley had 10 carries for 26 yards and one catch for 1 yard in his return during a 27-13 loss to the Denver Broncos on Sunday. He played 28 snaps -- 48% of the Giants' offensive plays -- and was mostly a nonfactor without the ability to get into open space.