Skipper Stiliyan Petrov, central defender Richard Dunne and left-back Stephen Warnock all suffered setbacks during Wednesday night's 2-0 success at Hull.
Dunne picked up a head injury in a clash with Tigers striker Jan Vennegoor of Hesselink although he was able to return after having it bandaged.
Warnock sustained a knock to his ankle, although he managed to complete the 90 minutes, and midfielder Petrov came off six minutes from time with a groin injury shortly after Villa's second goal.
Villa manager Martin O'Neill said: "I think Richard's cut may have reopened (after stitches) but I'm hoping he is going to be okay.
"Stephen was kicked twice in the same spot within a matter of moments but he carried on to the end.
"Stiliyan had a kick in the groin which was very sore at half-time but he stayed on until close to the end. We'll see how they are."
Villa need to overcome their nearest rivals to sustain their late push for a Champions League spot after collecting 10 points from their last four games.
They will also be looking for a fourth successive victory against them under O'Neill.