The Serie A powerhouse have named former Azzurri boss the veteran manager to lead the team for the remainder of the campaign.
The 66-year-old takes over from his predecessor, who was let go post the Lazio setback last weekend.
Tudor arrived at the club in March filling the shoes of Thiago Motta but left after seven months after an eight-game winless streak.
Spalletti guided Napoli to the Serie A crown in 2022-23 before taking charge of Italy, who had failed to qualify for the 2022 World Cup under former boss Roberto Mancini.
He helped them qualify for Euro 2024 but were eliminated in the round of 16 by Switzerland.
However, Spalletti was sacked by Italy in June following a loss to Norway in World Cup qualifying, putting Italy in danger of missing a third straight World Cup.
Spalletti's managerial career includes Inter Milan, Roma, and Udinese in Serie A and won the Russian Premier League twice with Zenit St Petersburg.
Massimo Brambilla served as interim manager for Juventus in the midweek 3-1 triumph over Udinese which left them seventh in Serie A - six points behind leaders Napoli after nine games.
In the Champions League, they sit 25th in the league phase, with one loss and two draws in their first three games.