Table 1.
Individual information.
Fig 1.
Positive control of immunohistochemical staining.
A: A positive area is found in the dermis using the mammary gland as a positive control for type 1 collagen. B: A positive area is found in the intracellular area of the mesothelial area using the normal canine aorta, selected as the positive control for elastin.
Fig 2.
The transition point (yellow dot) has been set at the intersection of the toe-region approximation line and the liner-region approximation line following the creation of the stress-displacement curve.
Fig 3.
Photograph used to measure the area of the femoral side in Image J.
The cranial cruciate ligament was transected perpendicular to the longitudinal axis of ligament with a scalpel at its attachments to the femur and tibia, and the section was stained with India ink to measure the cross-sectional area using Image J. The photograph was taken while the scale was taken to calibrate the object’s length.
Fig 4.
Hematoxylin-eosin staining findings of the cranial cruciate ligament in the GC and control groups (bar = 20 μm).
The upper panels represent examples of staining on the tibial (A), middle (B), and femoral (C) sides of the GC group. The lower panels represent examples of staining on the tibial (D), middle (E), and femoral (F) sides of the control group. In the box-and-whisker plot, blue, orange, gray, and yellow indicate the tibial side, middle, femoral side, and the entire cell count, respectively. If the plots are connected by a line, they are stifle joints of the same dog. Within each group, the interfascicular area and fibrocartilage are more extensive at the ligament attachments, particularly on the tibial side, than in the middle. Irregularities are observed in fibril orientation. In the analysis of cell density, there are no significant differences between the areas within each group and between the groups. GC, glucocorticoid.
Fig 5.
Alcian-blue staining findings of the cranial cruciate ligament in the GC and control groups (bar = 20 μm).
The upper panels represent examples of staining on the tibial (A), middle (B), and femoral (C) sides of the GC group. The lower panels represent examples of staining on the tibial (D), middle (E), and femoral (F) sides of the control group. In the box-and-whisker plot, blue, orange, gray, and yellow indicate the tibial side, middle, femoral side, and the entire measurement, respectively. The asterisks indicate P<0.05. If the plots are connected by a line, they are stifle joints of the same dog. Areas with positive Alcian-blue staining are particularly obvious in the areas of irregular fibril orientation around the round cells, such as the interfascicular area and fibrocartilage. Within each group, the positive Alcian-blue area ratio is significantly higher on the tibial side than on the femoral side and in the middle. The ratio is significantly higher in the control group than in the GC group on the tibial side. GC, glucocorticoid.
Fig 6.
Elastica-Eosin staining findings of the cranial cruciate ligament in the GC and control groups (bar = 20 μm).
The upper panels represent examples of staining on the tibial (A), middle (B), and femoral (C) sides of the GC group. The lower panels represent examples of staining on the tibial (D), middle (E), and femoral (F) sides of the control group. In the box-and-whisker plot, blue, orange, gray, and yellow indicate the tibial side, middle, femoral side, and the entire measurement, respectively. The asterisks indicate P<0.05. If the plots are connected by a line, they are stifle joints of the same dog. Positive areas of Elastica-Eosin are observed, particularly in the interfascicular area and fibrocartilage. In the control group, the ratio is significantly higher on the tibial side than on the femoral and middle. The ratio is significantly higher in the control group compared to the GC group in the tibial, middle, femoral, and entire areas. GC, glucocorticoid.
Fig 7.
Immunohistochemical staining of type 1 collagen findings of the cranial cruciate ligament in the GC and control groups (bar = 20 μm).
The upper panels depict examples of staining on the tibial (A), middle (B), and femoral (C) sides of the GC group. The lower panels depict examples of staining on the tibial (D), middle (E), and femoral (F) sides of the control group. In the box-and-whisker plot, blue, orange, gray, and yellow indicate the tibial side, middle, femoral side, and the entire measurement, respectively. If the plots are connected by a line, they are stifle joints of the same dog. Extensive positive regions are found in areas with normal ligament orientation. There are no significant differences between the areas within each group and between the groups in the positive ratio analysis. GC, glucocorticoid.
Fig 8.
Immunohistochemical staining of the elastin findings of the cranial cruciate ligament in the GC and control groups (bar = 20 μm).
The upper panels depict examples of staining on the tibial (A), middle (B), and femoral (C) sides of the GC group. The lower panels depict examples of staining on the tibial (D), middle (E), and femoral (F) sides of the control group. In the box-and-whisker plot, blue, orange, gray, and yellow indicate the tibial side, middle, femoral side, and the entire measurement, respectively. The asterisks indicate P<0.05. If the plots are connected by a line, they are stifle joints of the same dog. In the control group, the ratio is significantly higher on the tibial side than on the femoral and middle. The ratio is significantly higher in the control group than in the GC group on the tibial side. GC, glucocorticoid.
Fig 9.
Failure pattern of the biomechanical test.
A: The femoral side after the failure. The end of the ligament is attached to the bony fragment of the tibia (red circle). B: The tibial side after the failure. The cranial cruciate ligament is detached over the entire area (red circle).
Fig 10.
Tensile test results of the bone-ligament-bone model in the GC and control groups.
A: stress at the transition point; B: strain at the transition point; C: elastic modulus; D: maximum tensile strength; and E: maximum stress. The asterisks indicate P<0.05. If the plots are connected by a line, they are stifle joints of the same dog. The stress at the transition point is significantly lower in the GC group than in the control group. GC, glucocorticoid.