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Background: Oral submucous fibrosis (OSMF), a well-recognized oral potentially malignant disorder, results due to increased collagen production and reduced collagen degradation. Aims and Objectives: To qualitatively compare the staining properties of collagen in OSMF using two special stains based on their birefringent property using polarizing microscopy. The study also assessed the distribution and orientation of collagen fibers in different grades of OSMF. Materials and Methods: A total of 73 subjects with different clinical and histopathological staging of OSMF comprised the study population. Histopathological examination was done using hematoxylin and eosin stain, Van Gieson and picrosirius red. Collagen fibers were analyzed for polarization colors, distribution, and orientation. Results: Picrosirius red stained both thick and thin collagen fibers. Irrespective of the histopathological grades reddish orange and yellowish orange were the most predominant colors. Parallel arrangement of fibers was observed when stained with Van Gieson but picrosirius red stained sections showed a majority of parallel type I fibers with perpendicular type III fibers which increased with advancement in the histopathological grade. Yellowish orange and greenish yellow fibers were predominant in the lamina propria, while reddish orange fibers were predominant in the submucosa. Conclusion: Picrosirius red was found to be a better stain. Histopathological grading and polarization colors showed no association with each other. Collagen fibers were more thickly and tightly packed in the submucosa indicating that the process of fibrosis began there. The increase in perpendicular type III fibers with advancing histopathological grades suggested their role in fibrosis.
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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.
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Background: Oral submucous fibrosis (OSMF), a potentially malignant oral disorder has the highest rate of malignant transformation of about 7-13%. The connective tissue changes that occur in this disease are characteristic and are stained with special stains. Objective: The study was done to compare common and special stains under light microscopy and polarizing microscopy to evaluate the levels of fibrosis in oral submucous fibrosis and assess the type of collagen present in the stromal area. Materials and Methods: Fifty tissue blocks were selected from the archives and were prepared and stained with H&E, Masson's trichrome, Van Gieson and Picrosirius red and studied under light microscope and polarizing microscope respectively. Results: H and E stained slides were useful in diagnosing the lesion but was not able to highlight the level of fibrosis. Masson's trichrome and Van Gieson stained slides showed the depth of the lesion which extended even to the deeper muscle layer. The type of collagen present was definitively seen by the birefringence in polarizing microscopic study. Interobserver variation was less and all the values regarding the effectiveness of the special stains in detecting the level of fibrosis were statistically significant. Conclusion: Special stains can be used routinely in laboratories to demonstrate connective tissue lesions especially in cases of OSMF. Depth of the lesion and the area of involvement help in treatment planning to be delivered. Large scale studies with more categories and inclusion criteria are required along with the special stains to assess the other alterations in OSMF
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Objectives: Morphologic detection of connective tissue fiber changes in oral squamous cell carcinoma (OSCC) using special stains remains less documented. The aims of the present study were to study the collagen and elastic fibers in different stages of OSCC and to correlate these changes with two grading systems ‑ Broder’s and Bryne’s. Study Design: Forty‑eight cases of OSCC were studied using hematoxylin and eosin, Verhoeff’s ‑ Van Gieson stain for elastic fibers and picrosirius red stain for collagen fibers. The changes were compared with all the grades of carcinoma. Normal mucosa was taken as control. Results: Statistical analysis using Chi‑square and ANOVA, showed significant association between the grades of carcinoma and extracellular matrix changes. Greenish‑yellow collagen fibers were found to be significantly increased in the poorly differentiated/Grade 3 cases (P < 0.0001) where as well‑differentiated/Grade 1 cases showed predominantly reddish‑orange and yellowish‑orange birefringence of collagen fibers. Chi‑square analysis showed a significant amount of fragmented pattern of elastic fibers in poorly differentiated OSCC (χ2 = 104.45, P = 0.009)/Grade 3 OSCC (χ2 = 94.81, P = 0.016). Conclusion: The study of the connective tissue stromal changes can be used as an adjunct to histological grading and thereby helping the surgeon to determine the amount of marginal clearance.
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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 .
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Objective Hermaphroditic planarians possess a very important position in the systematic evolutionary history of animal, as they are capacity of complete regeneration. Hence, the research on histological structure of autofluorescence has been carried out to provide a crucial insight into the developmental and regenerative biology. Methods Part of histological structure of planarian (Dugesia japonica) was revealed with HE method, Masson method and Van Gieson method. Their autofluorescence was observed with ultraviolet. There were six planarians in each stained group and the autofluorescence group. Results Epidermis, outer epidermis of pharynx, protonephridium, intestine, the photoreceptor cells and longitudinal nerve cords, all radiated blue autofluorescence. The epithelial dissociation side of copulatory bursa radiated yellow autofluorescence, its middle part radiated blue autofluorescence, its fundus side radiated weakly blue autofluorescence. Testis could hardly give off autofluorescence. Pigment cells of eyepot could not give off autofluorescence. Conclusion The research on configuration and autofluorescence of planarian eye may offer help for the study of origin and evolutionary law on eye of invertebrate.
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[Objective]To find a simple and direct feasible method to detect bone tissue vascularize from comparing advantage or disadvantage of ink intravascular Dabble-VG dyeing and other tissue dyeing in the tissue section capillary vessel display.[Methods]One side femoral artery of 6 normal goat was mixed with liquor composed of 10 percentink,10 percent formaldehyde and 20 percent mannitol,volume ratio in 7∶2∶1.Stopped femoral vein effluent was black.To get off goat tibia,10 percent formaldehyde was fixed with.EDTA decalcification,cedar oil douse,fiat 50 to 100 micrometer pachy-section and compound ink-VG dyeing to observe capillary vessel.[Results]Compound ink-VG dyeing ould display relations of capillary vessel and surrounding tissue.But in control group,single HE,Masson and Weigert Resorcinol-aniline red dyeing after-stain VG dyeing ould display few capillary vessel and surrounding tissue,and recognize difficultly.[Conclusion]Compound ink-VG dyeing can display capillary vessel of bone clearly,and can disclose relationships of surrounding tissue as well.This method can try on in tissue vascularization detect.