Italy will have to do without Leonardo Bonucci and Roberto Gagliardini for the Nations League games against Poland and Bosnia-Herzegovina.
Defender Bonucci is out with a muscular problem having revealed he had played on with an adductor injury during the second half of Juventus' 1-1 draw with Lazio prior to the international break.
The centre-back had not featured in Thursday's friendly against Estonia, a game the Azzurri won 4-0 to stretch their unbeaten run to 20 matches.
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"I wanted to be there at all costs, but unfortunately I can't," Bonucci said in an interview with Rai Sport on Saturday.