Cartilage attachment morphology of the fetal cruciate ligaments of the knee: an immunohistochemical study using human fetal specimens.

Fetal cruciate ligaments of the knee provide two types of cartilage attachments: to a cartilage fovea or a simple continuation to the perichondrium. To examine a difference in matrix substance between a ligament attachment to the fovea and another attachment to the perichondrium. We histologically observed 12 human fetal femurs in which the posterior (or anterior) cruciate ligament provided a fovea-type (or a perichondrium-type) attachment. Immunohistochemistry of matric substances (aggrecan, versican, tenascin-c) was performed. In the knees, aggrecan was consistently positive in any cartilage, versican was in the joint surface and tenascin-c in the perichondrium. In contrast to the femoral attachment, the anterior and posterior cruciate ligaments consistently continued to the perichondrium at the tibial attachment (versican-, tenascin+). In the femoral condyles, tenascin-immunoreactivity was seen in both of a fovea-type and a perichondrium-type attachments, but versican was not in both. During development of the cartilage fovea, the growing ligament seemed to push the perichondrium into the cartilage and, much or less, the tenascin-positive perichondrium was likely to be involved into the fovea.

Spatial Change of Cruciate Ligaments in Rat Embryo Knee Joint by Three-Dimensional Reconstruction.

This study aimed to analyze the spatial developmental changes of rat cruciate ligaments by three-dimensional (3D) reconstruction using episcopic fluorescence image capture (EFIC). Cruciate ligaments of Wister rat embryos between embryonic day (E) 16 and E20 were analyzed. Samples were sectioned and visualized using EFIC. 3D reconstructions were generated using Amira software. The length of the cruciate ligaments, distances between attachment points to femur and tibia, angles of the cruciate ligaments and the cross angle of the cruciate ligaments were measured. The shape of cruciate ligaments was clearly visible at E17. The lengths of the anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL) increased gradually from E17 to E19 and drastically at E20. Distances between attachment points to the femur and tibia gradually increased. The ACL angle and PCL angle gradually decreased. The cross angle of the cruciate ligaments changed in three planes. The primordium of the 3D structure of rat cruciate ligaments was constructed from the early stage, with the completion of the development of the structures occurring just before birth.

Three-dimensional reconstruction of rat knee joint using episcopic fluorescence image capture.

OBJECTIVE: Development of the knee joint was morphologically investigated, and the process of cavitation was analyzed by using episcopic fluorescence image capture (EFIC) to create spatial and temporal three-dimensional (3D) reconstructions. METHODS: Knee joints of Wister rat embryos between embryonic day (E)14 and E20 were investigated. Samples were sectioned and visualized using an EFIC. Then, two-dimensional image stacks were reconstructed using OsiriX software, and 3D reconstructions were generated using Amira software. RESULTS: Cavitations of the knee joint were constructed from five divided portions. Cavity formation initiated at multiple sites at E17; among them, the femoropatellar cavity (FPC) was the first. Cavitations of the medial side preceded those of the lateral side. Each cavity connected at E20 when cavitations around the anterior cruciate ligament (ACL) and posterior cruciate ligament (PCL) were completed. CONCLUSION: Cavity formation initiated from six portions. In each portion, development proceeded asymmetrically. These results concerning anatomical development of the knee joint using EFIC contribute to a better understanding of the structural feature of the knee joint.

Sox9-expressing precursors are the cellular origin of the cruciate ligament of the knee joint and the limb tendons.

Sox9 expression defines cell progenitors in a variety of tissues during mouse embryogenesis. To establish a genetic tool for cell-lineage tracing and gene-function analysis, we generated mice in which the CreERT2 gene was targeted to the endogenous mouse Sox9 locus. In Sox9(CreERT2/+) ;R26R embryos, tamoxifen activated Cre recombinase exclusively in Sox9-expressing tissues. To determine the suitability of this mouse line for developmental stage-specific gene recombination, we investigated the cellular origins of the cruciate ligaments of the knee joint and the limb tendons, in which precursor cells have not been defined. The cells in these tissues were labeled after tamoxifen treatment before or at the stage of chondrogenic mesenchymal condensation, indicating that ligament and tendon cells originated from Sox9-expressing cells and that cell fate determination occurred at mesenchymal condensation. This mouse line is a valuable tool for the temporal genetic tracing of the progeny of, and inducible gene modification in Sox9-expressing cells.

Postnatal maturation of tendon, cruciate ligament, meniscus and articular cartilage: a histological study in sheep.

Orthopaedic basic science data on immature skeletons are rare in the literature. Since the number of knee injuries in young humans is steadily increasing, studies on immature animals such as sheep, which can be used as model systems are becoming more and more important. However, no baseline data are available on physiologic and morphologic changes during growth in the relevant tissues. In the present study, histomorphometric changes in the tendon of the musculus flexor digitalis superficialis, the cranial cruciate ligament, the medial meniscus and the articular cartilage of the medial femoral condyle were identified in sheep between the ages of 1 and 40 weeks postnatally. Profound changes in tissue composition during growth could be observed. A high cellularity in the early postnatal period decreases to a constant lower level after 18 weeks. Similar changes during postnatal growth could be observed for blood vessel density. Also, staining of alpha-smooth muscle actin (SMA) and vascular endothelial growth factor (VEGF) steadily decreased. In contrast, the number of components of extracellular matrix steadily increased in all tissues. The age of 18 weeks seems to be a threshold after which the tissue composition of the observed structures remains constant in this species.

Early development of the cruciate ligaments in staged human embryos.

Investigations were carried out on 43 serially sectioned human embryos of developmental stages 18 to 23. The homogeneous interzone of the future knee joint is observed in embryos at stage 18. During stage 19 this interzone is differentiated into dense, intensively stained, peripheral parts, which are the primordia of menisci and the medial portion, in which the cruciate ligaments are formed. All structures of the interior of the knee joint are more clearly delineated during stage 20, and they are well developed during the last embryonic week (stages 21-23).

Development of the human knee joint ligaments.

BACKGROUND: Many studies have been published on the development of the human knee joint, but scant attention has been given to the development of the knee joint ligaments. The only elements that have received much attention are the cruciate ligaments and their relationships with the synovial membrane. METHODS: We summarize our observations on the development of the knee joint ligaments in 50 serially sectioned human embryonic and fetal lower limbs (26 embryos and 24 fetuses). RESULTS: The patellar ligament begins to form in O'Rahilly stage 20, with the muscle fibers of the quadriceps muscle being attached inferiorly to the tibial tuberosity. The cruciate ligaments (beginning with the posterior) arise from the articular interzone in O'Rahilly stage 21. Subsequently, with the organization of the Wrisberg's meniscofemoral ligament, in week 10 of development, the cruciate ligament system is completed. The lateral collateral ligament begins to form in O'Rahilly stage 23, and from its first appearance it is independent of the knee joint capsule. At this time, development of the tendon of the popliteus muscle begins. The medial collateral ligament begins to develop in week 9 of development as a condensation of the joint capsule. Two weeks later, the intra-articular pad of fat begins to form from mesenchymal tissue below the patella and between the cruciate and the patellar ligaments. With the organization of the suprapatellar bursa in week 14 of development, knee joint development is complete. CONCLUSIONS: The morphogenetic time table of the knee joint ligaments was established.