Ryan Tannehill was impressive for the hosts, the quarterback throwing three touchdowns as he passed for 195 yards while rushing for 42 yards and another score.
Tennessee's recently-poor defence stepped up with two interceptions against in-form Buffalo signal-caller Josh Allen, who they also limited to 263 passing yards.
The Titans' Derrick Henry made the play of the night, and the one that sent broadcast commentators and those on Twitter into a frenzy, in the second quarter when the running back faced down cornerback Josh Norman.
Norman attempted to stop Henry from reaching a first down but was met with a stiff shove from the defending league rushing champion which sent the Buffalo player flying.
The Titans remain undefeated at 4-0 while the Bills suffered their first loss and will next take on the reigning Super Bowl winners the Kansas City Chiefs, who will be out for blood after falling 40-32 to the Las Vegas Raiders on Sunday.