The Ivory Coast international scored the winner in Chelsea’s 1-0 first-leg win in the semi-final clash, but is struggling with a knee injury.
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Drogba missed the London derby against Arsenal on Saturday, and Di Matteo told The Guardian: "At this moment of time, I don't know [whether the striker will be fit in time for the return with Barca]."
Di Matteo explained after his team's goalless draw at Arsenal that the whole Chelsea squad will be looked at ahead of the second leg with the Spanish giants.
"We'll have to assess the players over the course of the weekend. He [Drogba] has an injury on his knee, which is why he did not play here, so it's whether the progression on this injury will be quick enough [for him to feature]."
With Drogba sidelined, Di Matteo started with strikers Fernando Torres, Daniel Sturridge and Salomon Kalou against the Gunners.