Mathesius is a Czech surname. Notable people with the surname include:
Bohumil Mathesius (1888–1952), Czech poet, translator, publicist and literary scientistFrederick Mathesius, an American architectJohannes Mathesius (1504–1565), German-Bohemian minister and Lutheran reformerJohannes Mathesius, the Younger (1544, Jáchymov – 1607), Bohemian-German physicianPaul Mathesius (1548, Jáchymov – 1584), Bohemian-German Lutheran theologianVilém Mathesius (1882–1945), Czech linguistMathesius is also a surname of a Finnish family and may refer to:
Johan Mathesius (1709–1765), pastor and politician of the Caps partyPer Niklas Mathesius (1711–1772), pastor and politician of the Caps party, brother of Johan Mathesius