The Chelsea manager is set to rest the star duo along with several other key first-team players, following the weekend controversy involving referee Mark Clattenburg
The Stamford Bridge side have accused referee Clattenburg of using "inappropriate" language towards Mata and Mikel in Sunday's controversial 3-2 defeat to Sir Alex Ferguson's men, with both players expected to be kept out of the limelight on Wednesday night as the fallout intensifies.
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The Italian manager has suggested he will be resting a number of first-team players due to the Champions League holders' hectic schedule of fixtures.
"It is our fourth game in 10 days and they have all been big games, so we are looking to freshen up the team a little bit for sure and also put a couple of youngsters in," Di Matteo told reporters.
"We always talk about developing and giving youngsters a chance, so this game is a good opportunity for some of them."
The likes of Ryan Bertrand, Oriol Romeu, Daniel Sturridge and Marko Marin are likely to be given a chance to avenge Chelsea's league reverse at the hands of United, in which both Branislav Ivanovic and Fernando Torres were dismissed.
Javier Hernandez also scored the winner from what appeared to be an offside position with 15 minutes remaining, incensing the London club further, and the latest allegations over Clattenburg have inflamed the ongoing debate over racism in the modern game.