Poly(lactic acid) (PLA), compared to other biodegradable polymers, has one of the highest consumption volumes in the world. Due to its specific properties, especially those that make it ecologically acceptable, it has a wide application from food packaging to medical implants. In this work, commercially available 3D-printing filament of PLA was used. The objective was to investigate the effects of γ- radiation on structural changes, mechanical and thermal behaviour of the material. The samples were irradiated with total doses of 25 kGy, 50 kGy, 100 kGy and 200 kGy. The experimental techniques used to investigate the structural changes in PLA filaments were electron spin resonance (ESR) and gel permeation chromatography (GPC). Differential scanning calorimetry (DSC) and universal testing machine were applied in monitoring changes in thermal and mechanical behaviour, respectively. The formation and stability of free radicals were studied by ESR, which confirmed that the radical concentration depends on the applied absorbed dose, while the spectral lineshape appears to be dose independent. The loss of the ESR signal, directly related to the decrease of radical concentration, was monitored as a function of time for all applied absorbed doses. The breaking force and the Young’s modulus were determined and compared. Results have shown that both the concentration and decay rate constant of radicals depend on the absorbed dose. Compressive strength test, a new approach in considering the mechanical changes in irradiated PLA indicated a loss of material elasticity and an increase of brittleness by increasing absorbed dose.