The reigning champions have taken 21 points from their seven La Liga games, but will be without star attacker Messi - who has already scored eight league goals this term - due to a hamstring injury.
Gerardo Martino's men proved they can perform without their talisman in Tuesday's UEFA Champions League win at Celtic though, and they will look to do so again by extending their unbeaten home run to 25 Liga matches against a Valladolid side without a win in four.
Striker Diego Costa scored the only goal of the game as Diego Simeone's men secured their first league win over Real in 14 years, which they duly followed up with a 2-1 Champions League success at Porto.
Joaquin Caparros's side are unbeaten since their 7-0 hammering at the hands of Barcelona on the opening day, but they have not won in the league at Estadio Ciudad de Valencia since March.
Elche host Espanyol hoping to move up the table following their first win of the campaign over Celta, and two out-of-form sides meet when Real Sociedad take on Rayo Vallecano.
Elsewhere, Malaga face Osasuna, Almeria travel to Sevilla and Getafe meet Real Betis.
