Levi Reid and Michael Cummins will both miss the vital clash with MK Dons after the FA rejected a Vale appeal against Reid’s suspension.
Reid’s three match ban and Cummins’ one match ban will both be in place for this weekend’s visit to Milton Keynes.
Both players were controversially dismissed in the home match against Colchester which also saw the U’s Liam Chilvers shown red.
Robin Hulbert is still injured so the absence of Reid and Cummins could see either Steve Rowland or Craig James deployed in an emergency central midfield role alongside loanee Danny Sonner.
Vale appealed against the bans but their appeals were dismissed by the FA on Wednesday.
Milton Keynes will themselves be missing Craig Ewart who is serving a three match ban for a dismissal.