ABSTRACT
Objective Sema4D, as the earliest known immunosuppressive agent, may be involved in the development and progression of osteoporosis. This study aimed to investigate the relationship between sSema4D and osteoporosis.Methods The model of osteoporosis was established in 50 three-month-old C57 mice by gavage of 0.9% retinoic acid and another 50 mice were taken as controls, which received gavage of 0.9% sodium chloride, all at 90 mL/kg for 21 days. Micro-CT scanning of the proximal tibial metaphysis bone tissue 3D reconstruction were performed for observation of the structure, shape, arrangement, direction and number of the trabeculae. The CT values of the area of interest in the proximal tibia metaphysis were determined and the trabecular parameters were obtained with the Inveon Research WorkPlace software. The pathological changes of the bone collagen fibers in the femur and tibia were observed by Van Gieson staining, and the level of serum sSema4D was detected by ELISA.Results Compared with the controls, the osteoporosis model mice showed significant decreases in the trabecular number (7.0 mm vs 5.8 mm, P<0.05), thickness (0.08 mm vs 0.06 mm, P<0.05) and volume fraction (0.5% vs 0.3%, P<0.01), but a marked increase in the trabecular separation (\[0.54±0.12\] mm vs \[0.69±0.13\] mm, P<0.05). Van Gieson staining manifested scarce collagen fibers in the osteoporosis model mice, loosely and disorderly arranged, some poorly connected, dissolved and fractured, with the marrow cavity increased. The level of serum sSema4D was significantly higher in the osteoporosis group than in the control (P<0.01).Conclusion Serum sSema4D may contribute to the incidence of osteoporosis.
ABSTRACT
Objective To investigate whether and what staining techniques are applied to the ultrathin sheet plastination slice and whether the stained specimen is of autofluorescences .Methods A cadaveric hand block was plastinated and then sectioned as a series of 300-400μm thick transverse sections .A total of 56 slices in total .Alternative sections were stained with hematoxylin -eosin staining ( HE) , Verhoeff -Van Gieson staining ( VVG) or methylene blue and azureⅡstaining(MA).The stained slices were examined under a light microscope and a confocal microscope .Results The plastinated slices were stained with the three staining methods .HE staining revealed the muscle and connective tissues were red or violet , bone was violet or blue;VVG staining showed the elastic fibers was black , the collagen was red , and other tissues were yellow .MA staining showed the tendon was violet , the bone was pink , cartilage was violet , and other tissues were purple.However, the intracellular structures appeared not very well stained .The collagen, elastin and muscular structures in the stained slices were observed under a confocal microscope .Conclusion The commonly used histology staining methods can be used to stain the ultrathin sheet plastination slices .The staining provides a better observation of various tissues in the slice than the unstained slice .After staining, those autofluorescent structures in the plastinated slice are detectable under a confocal microscope .