Scans have confirmed that Gary Gardner suffered an anterior cruciate ligament injury in his right knee during the Professional Development League U21 game against Stoke on Wednesday.
The 20-year-old midfield player will undergo surgery and he faces being sidelined for eight to nine months.
"It's a blow and it's definitely a blow for the lad but he's very positive and he'll get great attention all the way through his rehab," said Villa manager Paul Lambert.
Gardner suffered an anterior cruciate ligament injury in his left knee in December 2009 but recovered and has made 17 appearances for the Villa first team, 12 as a substitute.
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