De Rossi was caught on camera punching Inter's Mauro Icardi and although the incident went unpunished at the time, Italy coach Cesare Prandelli has pre-empted a likely charge by dropping the player.
Prandelli is also without injured pair Mario Balotelli and Giuseppe Rossi, while Spain will hand an eagerly awaited debut to Atletico Madrid striker Diego Costa following his switch of allegiance from Brazil.
Spain defender Sergio Ramos said: "The World Cup will be here before we know it so the players and the fans are motivated. Italy is one of the biggest teams in the world."
Arjen Robben will sit out Holland's friendly in France with a thigh injury sustained in the process of scoring a hat-trick in Bayern Munich's Bundesliga win over Schalke at the weekend.
France coach Didier Deschamps will give Antoine Griezmann his debut in place of injured Franck Ribery as he looks to finalise his squad for Brazil. Deschamps said: "What matters are the values and potential my players show."
Miroslav Klose will become Germany's all-time leading goalscorer if he finds the next against Chile in Stuttgart.
Currently tied with Gerd Muller on 68 goals for his country, the 35-year-old will get his chance after recovering from injury while Bastian Schweinsteiger and Lukas Podolski also return to the squad.
England boss Roy Hodgson is desperate to avoid a third consecutive home loss as his side face Denmark at Wembley, with Alex Oxlade-Chamberlain hoping to get the chance to make an impression after a knee inury.
Portugal face fellow World Cup finalists Cameroon in Leiria, with Rafa and Ivan Cavaleiro set to get a chance to impress while Anthony Lopes starts in goal in place of Rui Patricio.
Belgium boss Marc Wilmots will move closer to finalising his first-choice line-up as his side face the Ivory Coast, with both Romelu Lukaku and Christian Benteke pushing for selection as his main striker.
Sweden look to put World Cup play-off woe behind them as they hand a debut to Fulham striker Muamer Tankovic in Turkey, while new Croatia boss Niko Kovac will look to experiment as his side face Switzerland in St Gallen.
Russia meet Armenia in Krasnodar, with Lokomotiv Moscow defender Maxim Belyaev and Spartak Moscow's Evgeny Makeev both added to Fabio Capello's squad.
Greece take on South Korea in Piraeus, while Ukraine's match against the United States is set to go ahead in Cyprus despite doubts over the fixture due to the current unrest in capital Kiev.
Brought back down to earth by their 4-1 friendly loss to the Faroe Islands at the weekend, UEFA new boys Gibraltar get a second chance to impress when they face Estonia in Portugal.
New Czech Republic coach Pavel Vrba has named no new players for his side's friendly against Norway in Prague, while Pambos Christodoulou takes charge of Cyprus for the first time against Northern Ireland.
Borussia Dortmund striker Robert Lewandowski is an injury doubt for Poland as they prepare to face Poland in Warsaw.
Aaron Ramsey is ruled out for Wales against Iceland in Cardiff, while Ireland face Serbia in Dublin and Slovakia travel to meet Israel in Netanya.