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Background and Objectives: Previous research has shown associations between atrophy and fatty infiltration of the lumbar paraspinal musculature and low back pain (LBP). However, few studies have examined longitudinal changes in healthy controls and individuals with LBP without intervention. We aimed to investigate the natural variations in lumbar paraspinal musculature morphology and composition in this population over a 4-month period. Materials and Methods: Healthy controls and individuals with LBP were age- and sex-matched and completed several self-administered questionnaires. MRIs of L1-L5 were taken at baseline, 2 months, and 4 months to investigate cross-sectional area (CSA), along with DIXON fat and water images. A total of 29 participants had clear images for at least one level for all three time points. Means and standard deviations were calculated for the participant demographics. A two-way repeated measures ANOVA was performed to investigate CSA, fat signal fraction, and CSA asymmetry. Results: A total of 27 images at L3/L4, 28 images at L4/L5, and 15 images at L5/S1 were included in the final analysis. There were significant main effects of group for psoas CSA at the L3/L4 level (p = 0.02) and erector spinae (ES) CSA % asymmetry at the L3/L4 level (p < 0.001). There was a significant main effect of time for lumbar multifidus (LM) CSA % asymmetry at L4/L5 level (p = 0.03). Conclusions: This study provides insights into LM, ES, and psoas morphology in both healthy controls and affected individuals over a 4-month period without any intervention. Our findings suggest that psoas CSA at higher lumbar levels and CSA % asymmetry in general may be a better indicator of pathology and the development of pathology over time. Evaluating natural variations in paraspinal musculature over longer time frames may provide information on subtle changes in healthy controls and affected individuals and their potential role in chronic LBP.