In 1874 he earned his medical doctorate from the University of Vienna, where he was a student of Theodor Billroth (1829-1894). Afterwards, he remained in Vienna for several years as Billroth's assistant. In 1886 he became a professor of surgery at the University of Graz, and in 1895 was a professor at Charles University in Prague.
Wölfler is remembered for his work in gastrointestinal surgery, and his investigations involving the thyroid gland. On September 28, 1881, he performed the first gastroenterostomy, which occurred on a patient with pyloric obstruction. He is also credited with providing the first detailed description of postoperative tetany.