Before meeting Ecuador later this week, the Azzurri play their first fixture since qualifying for the upcoming European Championship at the end of last year.
Ultimately aiming to defend their title this summer, Italy have called up a relatively inexperienced squad, as Spalletti sifts through the alternatives to tried and tested players such as Ciro Immobile, who has been left at home.
While most of the men that edged out Ukraine to seal a spot in Germany are still present, several fresh faces may have a chance to impress at Florida's Chase Stadium.
Captain Gianluigi Donnarumma already has 60 caps to his name at the age of 25, but Tottenham Hotspur's Guglielmo Vicario and Atalanta BC goalkeeper Marco Carnesecchi are now vying to make their first senior start for La Nazionale.
Still uncapped, young wing-backs Raoul Bellanova and Andrea Cambiaso have shone for Serie A's two Turin clubs, while Udinese striker Lorenzo Lucca joins former Fruilani star Destiny Udogie in hoping for selection.
Lucca is one of seven forwards included, along with Aston Villa loanee Nicolo Zaniolo, Federico Chiesa, Riccardo Orsolini, Giacomo Raspadori, Mateo Retegui and Mattia Zaccagni; however, Sassuolo's Domenico Berardi was recently ruled out for the rest of the season by injury and Gianluca Scamacca has been dropped.
Assuming Spalletti sticks with his preferred 4-3-3 formation on Thursday, Orsolini and Zaniolo are in contention to replace Berardi on the right wing, with Chiesa and Raspadori favourites to occupy the other two positions up front.
Following the enforced withdrawal of Inter Milan defender Francesco Acerbi, who stands accused of racism during Inter's recent draw with Napoli, Gianluca Mancini has been called up as cover.
There will be a big battle for places at full-back - or wing-back, should Italy opt for a variation of 3-5-2 - as Bellanova, Giovanni Di Lorenzo and Matteo Darmian contest the right flank; Federico Dimarco and Udogie are both strong contenders on the left, while Cambiaso can fill in on either side.
Italy possible starting lineup: Donnarumma; Di Lorenzo, Scalvini, Bastoni, Udogie; Barella, Jorginho, Pellegrini; Orsolini, Raspadori, Chiesa