The Serie A powerhouse have appointed previous Italian national team manager Spalletti as their manager for the rest of the term.
The 66-year-old takes over from his predecessor, who was let go post the Lazio setback recently.
The former boss came on board in March as Thiago Motta's replacement but departed after seven months after an eight-game winless streak.
Spalletti led Napoli to the Serie A title in 2022-23 prior to assuming the Italy role, after the team's failure to reach the 2022 World Cup during Roberto Mancini's tenure.
He helped them qualify for Euro 2024 exiting in the last 16 stage against Switzerland.
Nonetheless, Italy dismissed Spalletti in June after a World Cup qualifying defeat by Norway, putting Italy in danger of missing a third straight World Cup.
He has previously coached Inter Milan, Roma, and Udinese in Italy's top flight and lifted the Russian league trophy twice while at Zenit St Petersburg.
Massimo Brambilla served as interim manager for Juventus during the 3-1 victory over Udinese on Wednesday which left them seventh in Serie A - trailing leaders Napoli by six points after nine matches.
In the Champions League, they sit 25th in the league phase, with one loss and two draws in their first three games.
