Objective: This study evaluated the effect of chemical disinfection and microwave irradiation on the surface hardness and roughness of two commercially available hard relining materials (Ufi Gel hard, GC Kooliner) and one denture base resin (Trevalon). Materials and Methods: A total of 72 specimens (n = 24) were divided into four groups. C: Not disinfected, Cl: disinfected with 4% chlorhexidine solution, Gl: disinfected with 2% glutaraldehyde solution, Mw: disinfected with microwave irradiation (650 W; 6 min). Hardness and roughness measurements were made after polymerisation, 1st day, 14th day and 28th day. Results: Ufi Gel hard showed an increased roughness after 1st day (P = 0.021) following chemical disinfection and GC Kooliner showed similar results after 14th day (P < 0.05). Microwave irradiation showed a significant increase in surface roughness value after 1st day (P < 0.05) for both Ufi Gel hard and GC Kooliner. Hardness of both Ufi Gel (12.131 to 7.333 VHN) and Kooliner (9.133 to 5.276 VHN) was significantly reduced by chemical disinfection, while microwave irradiation resulted in an increased surface hardness of Kooliner (from 9.126 to 12.713 VHN) and Ufi Gel hard (from 11.698 to 14.940VHN). Results for Trevalon were not significant for both the disinfection methods. Conclusions: Microwave irradiation increased the surface roughness and hardness of Ufi Gel hard and Kooliner, while chemical disinfection resulted in a decreased hardness and increased roughness of both hard relining materials. There was no effect of either of the disinfection methods on Trevalon.