Edinson Cavani, Christian Maggio and Goran Pandev start, while Rolando makes his Napoli debut in the Europa League tie with Viktoria Plzen.
The first leg of this Round of 32 kicks off at 18.00 GMT at the Stadio San Paolo and takes place on Cavani’s birthday.
Lorenzo Insigne had been pencilled in to play, but pulled out earlier today with flu symptoms.
Instead, Pandev and Cavani maintain the usual line-up, though Omar El Kaddouri steps in so Marek Hamsik can rest.
Maggio returns for his first appearance since undergoing surgery on a fractured hand, while January buy Rolando makes his Napoli debut in defence.
Czech side Viktoria Plzen qualified as winners of Group B and field a 4-2-3-1 system led by their top scorer Marek Bakos.
Napoli: De Sanctis; Gamberini, Rolando, Britos; Maggio, Donadel, Dzemaili, El Kaddouri, Zuniga; Pandev; Cavani
Napoli bench: Rosati, Campagnaro, Cannavaro, Hamsik, Inler, Mesto, Calaiò
Viktoria Plzen: Kozacik; Reznik, Cisovsky, Prochazka, Limbersky; Horvath, Darida; Rajtoral, Kolar, Kovarik; Bakos
Viktoria Plzen bench: Pavlik, Hejda, Zeman, Stipek, Tecl, Fillo, Adamov