Pirates' conditioning coach Franck Plaine joined defender Marshall Munetsi on the sidelines after they were both sent for an early shower in their Telkom Knockout final penalty shootout defeat to Baroka a month ago.
The pair are serving the final match of their suspension after also missing out on the Absa Premiership win over Free State Stars in the club's last domestic assignment in December.
Plaine usually takes the players through their warm-up routines on matchday, but will have to relinquish his duties at Makhulong Stadium this Saturday evening.
Highlands will also miss the services of top goal scorer and highly sought-after midfielder Mothobi Mvala, as well as right-back Phoka Mofokeng.
Others serving suspensions include Bidvest Wits duo Thabang Monare and S'fiso Hlanti, who will be absent during their side's visit to Free State Stars.
Ea Lla Koto, meanwhile, will be without Nyiko Mobbie, Sibusiso Masina and Wits-bound midfielder Sinethemba Jantjie.
Maritzburg United will visit Golden Arrows minus Rushine De Reuck, while Chippa United have to make do without goalkeeper coach Brian de Heever who was banished from the bench against Wits last year.