This research aimed to identify changes in muscle mechanical properties (MMPs) when a standardized sequence of movements is performed and to determine the influence of acute low back pain (LBP) and age on the MMPs. Socio-demographic, clinical variables and MMPs were collected in 33 patients with LBP and 33 healthy controls. A 2 × 2 × 2 (group × age × time) analysis of variance (ANOVA) mixed model was used to determine the effect of the study factors on the different MMPs. There were no significant triple interactions. After the movements, tone and stiffness increased 0.37 Hz and 22.75 N/m, respectively, in subjects <35 years, independent of their clinical status. Relaxation showed differences by age in healthy subjects and creep in LBP subjects. Furthermore, elasticity was higher in <35 years (p < 0.001) without the influence of any other factor. In conclusion, sequenced movements can modify tone and stiffness as a function of age, while age-associated changes in viscoelastic characteristics depends on pain but not on movements. The MMPs should be assessed, not only at the beginning of the physical examination at rest, but also along the patient’s follow-up, depending on their pain and age, in a clinical setting.