Utilizing Fourier Transform Infrared Spectroscopy (FTIR) in specular reflectance mode chemical changes of root cement surfaces due to laser radiation were investigated. A total of 18 samples of root cement were analyzed, six served as controls. In this study laser energies were set to those known for removal of calculus or for disinfection of periodontal pockets. Major changes in organic as well as inorganic components of the cementum were observed following Nd:YAG laser irradiation (wavelength 1064 nm, pulse duration 250 μs, free running, pulse repetition rate 20 Hz, fiber diameter 320 μm, contact mode; Iskra Twinlight, Fontona, Slovenia). Er:YAG laser irradiation (wavelength 2.94 μm, pulse duration 250 μs, free running, pulse repetition rate 6 Hz, focus diameter 620 μm, air water cooling 30 ml/min; Iskra Twinlight, Fontona, Slovenia) significantly reduced the Amid bands due to changes in the organic components. After irradiation with a frequency doubled Alexandrite laser (wavelength 377 nm, pulse duration 200 ns, q-switched, pulse repetition rate 20 Hz, beam diameter 800 μm, contact mode, water cooling 30 ml/min; laboratory prototype) only minimal reductions in the peak intensity of the Amide-II band were detected.