NiO thin films with various strains were grown on SrTiO 3 (STO) and MgO substrates using a pulsed laser deposition technique. The films were characterized using an x-ray diffraction, atomic force microscopy, and infrared reflectance spectroscopy. The films grown on STO (001) substrate show a compressive in-plane strain which increases as the film thickness is reduced resulting in an increase of the NiO phonon frequency. On the other hand, a tensile strain was detected in the NiO film grown on MgO (001) substrate which induces a softening of the phonon frequency. Overall, the variation of in-plane strain from −0.36% (compressive) to 0.48% (tensile) yields the decrease of the phonon frequency from 409.6 cm −1 to 377.5 cm −1 which occurs due to the ∼1% change of interatomic distances. The magnetic exchange-driven phonon splitting Δω in three different samples, with relaxed (i.e. zero) strain, 0.36% compressive strain and 0.48% tensile strain, was measured as a function of temperature. The Δω increases on cooling in NiO relaxed film as in the previously published work on a bulk crystal. The splitting increases on cooling also in 0.48% tensile strained film, but Δω is systematically 3-4 cm −1 smaller than in relaxed film. Since the phonon splitting is proportional to the non-dominant magnetic exchange interaction J 1 , the reduction of phonon splitting in tensile-strained film was explained by a diminishing of J 1 with lattice expansion. Increase of Δω on cooling can be also explained by rising of J 1 with reduced temperature.