Guardino was born to an Italian family on the Lower East Side of Manhattan and raised in Brooklyn, New York. He joined a Police Athletic League dramatic group while attending high school. After graduation, Guardino joined the Navy, serving in World War II. After the war, he became a merchant seaman. Guardino appeared on stage, in films, and on television. His Broadway theatre credits included A Hatful of Rain, One More River (earning a Tony Award nomination for Best Featured Actor in a Play for his performance), Anyone Can Whistle, The Rose Tattoo, The Seven Descents of Myrtle, and Woman of the Year.
Guardino's other film credits include Houseboat, Pork Chop Hill (about the Korean War), The Five Pennies, King of Kings, Madigan, Lovers and Other Strangers, Dirty Harry and The Enforcer. He was nominated twice for the Golden Globe Award for Best Supporting Actor. He guest starred on John Cassavetes's 1959–1960 series, Johnny Staccato, the story of a pianist/private detective in New York City.
In 1960, Guardino appeared as Johnny Caldwell in the episodes "Perilous Passage", "The O'Mara's Ladies", and "Daughter of the Sioux" in the NBC western series Overland Trail starring William Bendix and Doug McClure.

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