Argentine Guillermo Canas also withdrew on Thursday after undergoing surgery also on a wrist injury, the USTA reported.
Safin, who would have been the 26th seed, has only played once since April. However he was unable to finish that first round match in Los Angeles against India's Prakash Amritraj last month because of the injury.
The 23-year-old Russian won his only Grand Slam title at the US Open in 2000, defeating Pete Sampras in the final. Sampras is to announce his retirement on the first day of the tournament next Monday.
Canas, once Argentina's highest ranked player and 12th in the world a year ago, was making a comeback at the tournament after not playing since mid-January because of his injury.
He has dropped to 209th and was to play Spain's French Open champion, and tournament third seed, Juan Carlos Ferrero in the first round.
Both players will be replaced by lucky losers -- the highest ranked players who lose in the final round of qualifying.
