Former World Cup winner Fabio Grosso has missed out on the finals this time.
Italy coach Marcello Lippi has reduced his 30-man provisional group to 28 and the Juventus defender and Udinese midfielder Antonio Candreva are the two to be axed.
Grosso, 32, was a pivotal player for the Azzurri four years ago when Lippi's side lifted the trophy.
He scored the first goal against Germany in the semi-final and then the winning spot-kick in the penalty shootout against France in the final.
Candreva, meanwhile, made his debut with the national side last November in a friendly against Holland.
But the 23-year-old, who played alongside Grosso while at Juve last season, has made just one appearance for Italy since then.
The Azzurri will train in Sestriere from Sunday onwards and Lippi will cut his squad to the final 23 by 1st June.
They will then play two friendly internationals against Mexico on 3rd June in Brussels and against Switzerland two days later as they round off their World Cup preparations.
Italy squad: Gianluigi Buffon (Juventus), Morgan De Sanctis (Napoli), Federico Marchetti (Cagliari), Salvatore Sirigu (Palermo); Gianluca Zambrotta (Milan), Fabio Cannavaro (Juventus), Giorgio Chiellini (Juventus), Christian Maggio (Napoli), Leonardo Bonucci (Bari), Domenico Criscito (Genoa), Mattia Cassani (Palermo), Salvatore Bocchetti (Genoa); Andrea Pirlo (Milan), Gennaro Gattuso (Milan), Daniele De Rossi (Roma), Riccardo Montolivo (Fiorentina), Mauro Camoranesi (Juventus), Angelo Palombo (Sampdoria), Andrea Cossu (Cagliari), Claudio Marchisio (Juventus), Simone Pepe (Udinese); Fabio Quagliarella (Napoli), Giuseppe Rossi (Villarreal), Vincenzo Iaquinta (Juventus), Antonio Di Natale (Udinese), Marco Borriello (Milan), Alberto Gilardino (Fiorentina), Giampaolo Pazzini (Sampdoria).