The Chelsea boss admitted after the game he would consider dropping the Spain striker but said on Monday: "I'm not going to disclose my ideas about tomorrow [Tuesday].
Di Matteo also hinted at the weekend he was prepared to abandon the swashbuckling style that has earnt Chelsea so many plaudits this season in order to secure a much-needed result - an approach that paid off spectacularly last term.
Most of all, Di Matteo needs his players to react positively to Saturday's dressing down.
"There was obviously frustration for the result on Saturday," he said.
"It's normal when things don't go your way.
"We owe ourselves a good performance and anything can happen.
"But we've proven many times before that, when it counts, our players can be counted for.
"Everyone is fully committed for tomorrow [Tuesday]."
That loss heralded the first visible cracks in the Chelsea dressing room, with players said to have argued loudly and Di Matteo also thought to have been furious with their performance.
But the Italian "absolutely" welcomed the frank exchange, saying: "It's important that we communicate with each other, and the players express themselves. I've always encouraged that."
Crucially, there is no obvious sign of Di Matteo having lost the confidence of his players in the way that cost Villas-Boas so dearly.
He added: "They need to have belief in themselves, and that's the most important aspect.
"We work together and, at the moment, we're all in it together and believe we have a good group, a good team, and are pulling together."
Having been especially critical of a defence that went a ninth game without a clean sheet on Saturday, Di Matteo knew the challenge facing his side on Tuesday in the absence of the injured John Terry.
"We need to pay attention and focus in certain situations defensively," he said, revealing Ashley Cole would be back from a hamstring problem.
"We have enough players to deal with that, but the focus will be very important for that."
So finely poised is Group E that a Chelsea win tomorrow would secure qualification, while a draw would leave them needing only a routine victory against Nordsjaelland to progress.
A defeat, however, and Juventus would require only a draw at Shakhtar Donetsk - who could well have little to play for - to reach the last 16 at Chelsea's expense.
Di Matteo said: "We know the importance of the game tomorrow [Tuesday].
"Motivation is going to be very, very high.
"It's important we understand that and show, as I said, the commitment that's necessary."