The centre-half had recently returned to training after a third operation of the season on a niggling groin problem.
But Villa boss Paul Lambert confirmed the 33-year-old, who is out of contract in the summer, has sustained another blow to his hopes of a first-team comeback.
Lambert said: "It looks like Richard has got a setback. We will have to wait and see if that settles down again in the next few days.
"He has been training but came in one morning this week and felt something."
Lambert had hoped Dunne could play some part in Villa's relegation scrap, despite having made only a handful of appearances in the last 12 months.
He suffered a shoulder injury which forced him to miss several games at the end of last season and then picked up the groin injury when on Euro 2012 duty for the Republic of Ireland.
Lambert also revealed midfielder Fabian Delph is sidelined with an ankle
He said: "It's the same ankle he hurt against West Brom and Millwall but hopefully he might train at the end of the week."