Micro- and Ultrastructural Characterization of Age-Related Changes at the Anterior Cruciate Ligament-to-Bone Insertion

There remains a lack of understanding of the structural changes that occur across the complex, multitissue anterior cruciate ligament (ACL)-to-bone insertion as a function of aging. The objective of this study is to provide a multiscale comparison of matrix properties across the skeletally immature and mature ACL-to-bone insertion. Using complementary imaging methods, micro- and ultrastructural analysis of the insertion revealed that collagen fiber orientation at the interface changes with age, though the degree of collagen organization is maintained over time. These changes are accompanied by a decrease in collagen fibril density and are likely driven by physiological loading. Mineral crystal structure and crystallinity are conserved over time, despite regional differences in crystallinity between the interface and bone. This suggests that mineral chemistry is established early in development and underscores its important functional role. Collectively, these findings provide new insights into interface development and set critical design benchmarks for integrative soft tissue repair.