Matsui started as the designated hitter, batting seventh against the Toronto Blue Jays.
"Our plan is to play him every day," manager Joe Girardi said. "Obviously, we'll evaluate him every day he gets in here and see how his knee is holding up."
Matsui, who is batting .323 with seven homers and 34 RBIs in 69 games, has been out since June 22. His sore left knee flared up several times during the rehab, and he decided not to have surgery last month in an attempt to rejoin the team this season. Matsui isn't likely to play the outfield again this season.
"It would have to be quite an emergency," Girardi said.
Matsui had 15 at-bats during a simulated game at the Yankees' spring training complex in Tampa, Fla., on Sunday, facing left- and right-handed pitching.
The Yankees are still deciding whether Carl Pavano will pitch for them this weekend.
The oft-injured righty is in the mix to start Saturday at Baltimore. Pavano has been in just two games for New York since 2005.
"There's still internal discussions about what we're going to do and I should have something for you before we go to Baltimore," Girardi said.
"There's some other people that are possible," he said. "We have to wait and see how some things come out and then I'll give it to you on Thursday."
Victor Zambrano, who spent last year in the Toronto, Pittsburgh and Baltimore systems, signed earlier this season with the Yankees and could also be considered.
The 32-year-old Pavano signed a $39.95 million, four-year contract with the Yankees before the 2005 season but injuries to his shoulder, back, buttocks, elbow and ribs have limited him to only 19 appearances for New York. When healthy, his record is 5-6.
On the disabled list all season, Pavano started a minor league rehab assignment less than a month ago and has gone 1-1 with a 3.32 ERA in five starts with Class-A Charleston of the South Atlantic League and Double-A Trenton of the Eastern League.
In his last outing, he allowed one earned run and six hits in six innings Sunday at Reading, striking out six and walking one.
Hughes started at Triple-A on Sunday and allowed five runs and eight hits in 3 2/3 innings.
Chamberlain, who went on the disabled list Aug. 6 with rotator cuff tendinitis, threw in the outfield for the second straight day Tuesday. He'll take Wednesday off and play catch again Thursday, then throw a bullpen session in Baltimore on Saturday.
"As of now, everything feels good," Chamberlain said. "It feels a lot better than it has, which is a good sign. It's coming out good. The more I play catch, the better it feels."
The 22-year-old Chamberlain said he hopes to be activated by early September, but Girardi refused to put a timeline on the second-year right-hander.
"I just have it up until Saturday and we'll see how he feels come Sunday morning," Girardi said.