High Al content AlxGa1-xN/GaN multiple quantum well (MQW) films with different interlayers were grown by metal organic chemical vapor deposition. These MQWs were designed to achieve intersubband (ISB) absorption in the mid-infrared spectral range. We have considered two growth conditions, with AlGaN interlayer and GaN/AlN superlattice (SL) interlayer, both deposited on GaN-on-sapphire templates. Atomic force microscopy images show a relatively rough surface with atomic-step terraces and surface depression, mainly dominated by dislocations. High-resolution X-ray diffraction and transmission electron microscopy analyses indicate that good crystalline quality of the AlGaN/GaN MQW layer could be achieved when the AlGaN interlayer is inserted. The ISB absorption with a peak at 3.7 μm was demonstrated in MQW films with AlGaN interlayer. However, we have not observed the infrared absorption in MQW films with GaN/AlN SL interlayer. It is believed that the high dislocation density and weaker polarization that resulted from the rough interface are determinant factors of vanished ISB absorption for MQW films with the GaN/AlN SL interlayer.