This study aimed to characterize the mechanical properties of native human ligamentum flavum (LF) and correlate them with histopathological changes. Mechanical property gradients across the cranial, medial, and caudal regions of LF were mapped and compared with histological sections. We also compared lumbar spinal stenosis (LSS) samples with disc herniation (DH) samples as reference material to identify differences in mechanical properties and histopathological features. Our results revealed significant heterogeneity in LF mechanical properties, with local variations correlating with specific histopathological changes such as chondroid metaplasia and loss of elastic fibers. These findings underscore the importance of considering LF heterogeneity in mechanical characterization and provide insights into its behavior under pathological conditions.