The Portuguese international was replaced by Branislav Ivanovic after limping off in the 2-1 defeat at Aston Villa on Saturday.
And Ancelotti said the problem had prevented him from training for the past two days.
'Bosingwa had a problem with his knee in the game against Aston Villa and he will not play against Atletico Madrid,' he said.
'But it is not a big problem and we should be able to use him on Saturday.'
The Italian revealed better news on another of the Blues' star defenders, Alex, who is on the road to recovery after undergoing surgery on a groin injury in the summer.
The Brazilian played a reserve fixture at Brentford on Monday night, coming through with no ill effects, and Ancelotti says he could be back in the first-team fold by the weekend.
'Alex has to train for some days but he will be ready for Saturday,' the manager added.
Chelsea are looking to make it three straight wins in Europe this season against Atletico at Stamford Bridge, after opening their campaign with victories over FC Porto and Apoel Nicosia.
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