Lorenzo Pellegrini and Matteo Politano left the Italy camp with injuries on Tuesday ahead of the European champions' Nations League match with England.
Midfielder Pellegrini played on with thigh pain during Roma's 1-0 home loss to Atalanta at the weekend and he was sent home after tests at Italy's Coverciano training centre.
Politano was substituted with a sprained ankle shortly after scoring in Napoli's win over title rivals AC Milan on Sunday and coach Roberto Mancini decided not to risk further aggravating the injury.
The pair drop out a day after Paris Saint-Germain midfielder Marco Verratti was forced out of contention with a calf injury.
Mancini has called up Sassuolo's Davide Frattesi, Sampdoria forward Manolo Gabbiadini and youngster Salvatore Esposito, who plays for SPAL in Serie B.
Mancini's team hosts England at the San Siro on Friday and then travels to League A, Group C leaders Hungary three days later with a chance of qualifying for the final in March.
Italy sits third on five points, two behind Hungary and three ahead of winless England with two matches remaining.
Whoever wins the group makes it through to the final four, while the bottom team will be relegated to League B.
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