Enzymatic Elimination of Collagen in the Alcian Blue Staining of Connective Tissue Ground Substance

1961 ◽  
Vol 36 (2) ◽  
pp. 53-55 ◽  
Author(s):  
John R. Ring ◽  
Dorothy Krug ◽  
Ann Dally Ogawa
1997 ◽  
Vol 45 (8) ◽  
pp. 1157-1163 ◽  
Author(s):  
Weiqi Lin ◽  
Svetlana Shuster ◽  
Howard I. Maibach ◽  
Robert Stern

The apparent intensity of hyaluronan (HA) staining in tissue sections can vary as a function of fixation techniques. We examined the histochemical distribution of HA in normal human skin using an HA-specific binding peptide derived from bovine nasal cartilage. The HA, particularly in the dermis, was best preserved in sections fixed in 10% acid-formalin with 70% ethanol. In contrast, sections fixed in the routine 10% neutral-buffered formalin had a much weaker intensity of HA staining. Furthermore, acid-formalin/ethanol-fixed sections retained much of their apparent HA after incubation with saline, in contrast to the neutral formalin-fixed sections, in which most of the stainable HA was lost. Such marked differences in staining intensity were not observed in slides stained with Alcian blue, a procedure pressumed to stain HA as well as other glycosaminoglycans. Staining using the HA binding peptide was entirely absent when sections were first preincubated in hyaluronidase, whereas similar Alcian blue-stained sections retained most of their staining intensity. Caution should be exercised in evaluating the distribution of HA in tissues using the HA binding peptide, particularly when different fixation techniques among several laboratories are being compared. In addition, the ability to evaluate the HA content of tissues using Alcian blue staining should be reconsidered. The sulfated glycosaminolglycans of the “ground substance” appear to be the predominant substrates for Alcian blue. (J Histochem Cytochem 45:1157–1163, 1997)


2018 ◽  
Vol 46 (1) ◽  
pp. 8
Author(s):  
Gabriela De Bastiani ◽  
Flávio Desessards De La Corte ◽  
Karin Erica Brass ◽  
Camila Cantarelli ◽  
Stefano Dau ◽  
...  

Background: The injury repair process in tendons and ligaments includes different phases such as inflammation, neovascularization, fibroblast proliferation and fibrosis. Collagen type and tissue characteristics of tendon and ligament repair are described such as type collagen differentiation and properties of the scars tissue. The degeneration of articular cartilage when, characterized by loss of the articular layers associated of the decreased of proteoglycans. The aim of this study is to describe by histochemistry techniques the characteristics of tissue scar, collagen type in the repair process of tendons and ligaments, as well as articular cartilage degeneration.Materials, Methods & Results: Tissue samples of equine tendons, ligaments and articular cartilage of the metacarpophalangeal joint region were evaluated by ultrasonography, macroscopically and prepared for routine histopathology (H&E staining). The inclusion criterion of the samples in this study was based on the presence of lesions characterized in H&E stain as fibroplasia, neovascularization, collagenolysis, chondroid metaplasia in tendons and ligaments and fibrillation and cartilaginous eburnation lesions in the articular cartilage samples. The Masson’s trichrome, Picrosirius red and Alcian blue staining techniques were also performed in addition to H&E. Pathologic findings in the tendons and ligaments included fibroplasia, collagenolysis, chondroid metaplasia and lymphohistioplasmacytic inflammation. Tendons and ligaments scars were composed of type III collagen but there was also some type I collagen. Fiber alignment of tendons and ligaments in the reorganization tissue was not flawless and the fiber appearance was characterized by a lack of the fiber crimp and parallelism. The fibroplasia was characterized by endotendinous tickening areas associated with the presence of loose connective tissue. In the areas of loose connective tissue substitution, collagen type fibers are intercalated to a lesser extent by type-III collagen fibers. In the Alcian blue stained samples of articular cartilage observed the surface layer and the matrix zone of calcified cartilage were weakly stained in blue.Discussion: Three special stains were utilized in this study along with the H&E evaluation elucidating the behavior tendons, ligaments and articular cartilage injury. The important observation in this study was fibroplasia in tendons and ligaments seems to be composed by abundant of loose connective tissue, chondrocytes and intermingled collagen type I and III fibers associated with lack of crimps alignment of the fibers. The fragile structure suggested by the Masson’s trichrome stain results (presence of the loose connective tissue) in this study perhaps make the tendons and ligaments receptive to other lesions. The characteristic blue discoloration of collagen fibers was only observed in the loose connective tissue may be because the dye penetration becomes easier when compared to the dense connective tissue (stained in red). The Masson’s trichrome made possible the differentiated the dense connective tissue of the loose connective tissue. The combined histochemistry staining technics allowed an improved characterization of fiber alignment, collagen type, inflammatory cell infiltration and neovascularization, which happens during the repair process of tendons and ligaments. The fibrillation and eburnation of the articular cartilage were associated with the decrease Alcian Blue staining characterized by degeneration process of articular cartilage.


1967 ◽  
Vol 125 (3) ◽  
pp. 409-428 ◽  
Author(s):  
Betsy G. Bang ◽  
Frederik B. Bang

Infectious laryngotracheitis can be produced in chickens as an experimental model of severe nonfatal rhinitis and sinusitis. Inoculated intranasally into unanesthetized baby chicks it remains limited to the nasal fossa, produces acute desquamation of all nasal epithelia, results in functional recovery of the respiratory epithelium, but leaves important residual abnormalities. From the earliest recognizable lesions through 4½ months' convalescence, the principal changes are as follows: 1. Initial lesions, or small syncytia of intranuclear "inclusions", first identifiable in the mucociliated cells of the shallowest portion of the epithelium at about 21 hr postinoculum (the inner surface of the maxillary conchal scroll). 2. Acute sloughing, (about 3 to 7 days), marked by: (a) spread of lesions from cell to cell via multinucleated "giant cells" which progressively slough and desquamate respiratory, olfactory, and sinus epithelia, epithelial neural elements and blood vessels; (b) appearance of numbers of eosinophilic leukocytes along the basement membrane at the sites of lesions just previous to sloughing; intensive infiltration of the submucosa with small lymphocytes after sloughing begins; (c) histochemical change in the intracellular mucus of the cells which comprise the syncytia: this mucus stains with Alcian blue alone when stained with AB-PAS; and (d) all cartilages of the maxillary conchae become flaccid, and the cell nuclei and matrix lose both basophilic and Alcian blue staining properties, effects which recede by about the 8th day. 3. Repair (about 8 to 21 days), marked by rapid initial spread of a sheet of epithelial cells over the infiltrated subrmucosa, appearance of numbers of plasma cells circulating in the tissues, formation of encapsulated secondary nodules, and mucosal adhesions. 4. Convalescence (about 1 to 4½ months when experiments terminated), marked by functional restoration of the mucociliary lining of the nasal fossa. However, at 4½ months eight specimens all show complete metaplasia of the olfactory organ (end nerves, supporting cells, and glands of Bowman) to mucociliated epithelium, all show abnormal formation and alignment of mucous acini, and about 50% have severe persistent sinusitis.


Parasitology ◽  
1969 ◽  
Vol 59 (3) ◽  
pp. 505-518 ◽  
Author(s):  
R. E. Howells ◽  
D. A. Erasmus

Regional differences in the tegumentary tissue of Moniezia expansa, as revealed at the light-microscope level by histological and histochemical techniques, are described and evidence for secretory activity by the interproglottidal glands is presented.In very immature proglottides the interproglottidal glands are at the ‘precryptic’ stage. Gland cells may be differentiated from other tegumentary cells by their high RNA content and in certain gland cells the presence of an alcian blue staining material.In mature proglottides the glands consist of rosette-like clusters of cells around crypt-like intuckings of the tegument. Two types of cells are found in the gland, small alcian blue-staining cells which are most numerous in the neck region of the crypt, and larger cells, the predominant gland cells, which do not stain with alcian blue but possess non-specific esterase activity. No other tegumentary cells in Moniezia exhibit this activity. Esterase and phosphatase activity is found in the tegument and crypt of the glands and in the interproglottidal folds.The non-enzyme histochemistry confirms and extends the observations of previous workers.Cytochrome oxidase and succinic dehydrogenase were detected in the tegumentary cells and tegument. Very strong reactions were given in the neck and scolex, with a progressive diminution of activity posteriorly along the strobila. Very low activities were recorded in the tegument of the glands.


1997 ◽  
Vol 110 (21) ◽  
pp. 2691-2701 ◽  
Author(s):  
N.S. Stott ◽  
C.M. Chuong

Members of the vertebrate hedgehog gene family (HH) are involved in patterning and modulation of differentiation. Recently it has been shown that ectopic expression of HH gene family members in vivo blocks chondrocyte maturation through activation of a parathyroid hormone related peptide (PTHrP) dependent negative regulatory loop in the perichondrium. However, the direct effect of HH on chondrocyte maturation has not been tested. Here, we studied the effect of retroviral overexpression of the chicken sonic hedgehog gene (Shh) on the growth and maturation of limb bud cells in micromass cultures. Shh is neither expressed nor required for the initiation of cellular condensation in normal micromass cultures. With Shh over-expression, micromass cultures developed novel tightly whorled nodules in addition to the normal Alcian Blue positive cartilage nodules. We characterized the new nodules and showed that they are strongly positive for alkaline phosphatase, enriched in type X collagen and weakly positive for Alcian Blue staining. Shh overexpression also increased cell proliferation, but this cannot account for the formation of the new nodules. This current study shows that misexpression of Shh in in vitro chondrogenic cultures promotes characteristics of hypertrophic chondrocytes. Thus HH has two complementary functions; a direct positive effect on chondrocyte hypertrophy in the absence of PTHrP pathway, and an indirect negative feedback loop through PTHrP to prevent other less differentiated chondrocytes from becoming hypertrophic. These two complementary actions of HH coordinate the progression of cartilage maturation.


2017 ◽  
Vol 25 (6) ◽  
pp. 533-535 ◽  
Author(s):  
Francesco Nesa ◽  
Luca Poggi ◽  
Stefano Ferrero ◽  
Alessandro Del Gobbo

Extensive extracellular mucin deposition is a rare pathological thyroid condition with 6 cases described in literature so far. We report another case of a 67-year-old woman, discussing histopathological features, and review the literature. Our findings showed a diffuse mucin deposition in the stromal compartment of thyroid parenchyma. Histochemical stainings showed positivity for Alcian blue staining, but not for periodic acid–Shiff staining. Our case is peculiar because this mucin deposition was associated with benign nodular hyperplasia, in contrast with the other 6 reports, which described the same stromal alterations associated with benign or malignant thyroid tumors.


2018 ◽  
Vol 7 (2) ◽  
pp. 57-59
Author(s):  
Rakhmiyati Rakhmiyati ◽  
Muhammad Ja’far Luthfi

Common House Gecko (Hemidactylus frenatus) is one of reptiles that have ability to autotomy their tails. Tail autotomy is a mechanism to protect it self from predators. After the tail broke, there will be wound healing on the tail which is then followed by a tail regeneration event. Original tail and regenerate tail is very different morphologically and anatomically. The original tail is composed of bones while the tail of the regenerate is composed of cartilage. Histochemical staining using Alizarin Red-S Alcian Blue was done to differentiate bone and cartilage. This method will stained bones red while the cartilage will stained blue.


2020 ◽  
pp. 153857442096611
Author(s):  
Alptekin Yasim ◽  
Erdinc Eroglu ◽  
Sezen Kocarslan ◽  
Atila Yoldas ◽  
Mehmet Acipayam ◽  
...  

Introduction: To study the efficiency of internal compression therapy (ICT), a new and promising method of treatment for deep venous insufficiency, how that efficiency is achieved, and its potential side-effects, in a porcine model. Material and Methods: The femoral vein diameters of 4 pigs were first measured. ICT was then applied such as to reduce the diameter of these veins by 50%. The femoral vein diameters of 2 pigs were re-measured after 1 month. The femoral vein and its surrounding tissue were excised for immunohistopathological and genetic examination. The same procedures were applied to the remaining 2 pigs 3 months subsequently. Collagen I and IV immunohistochemical staining and Masson’s trichrome and Alcian blue histochemical staining were applied during immunohistopathological examination. Collagen I, III, and IV and connective tissue growth factor (CTGF) mRNA expressions were examined for genetic examination. Results: The femoral vein diameters decreased by approximately 50% after ICT application. This decrease persisted after the first and third months. Histopathological examination revealed loose connective tissue around the venous tissue after the operation, particularly in the third month, together with perivascular fibrosis and increased collagen in connective tissue. No difference was observed between regions with and without ICT application in terms of mucinous degeneration, an indicator of tissue injury, during Alcian blue staining. Genetic examination revealed an increase in collagen I and IV and CTGF mRNA expression in perivascular tissue resulting from ICT application. Conclusion: ICT is effective both in terms of creating a durable tissue around the vein and of increasing collagen tissue and stimulating fibrosis, and has no deleterious side-effects on tissue.


1985 ◽  
Vol 10 (1) ◽  
pp. 49-53 ◽  
Author(s):  
MARIE A. BADALAMENTE ◽  
L. C. HURST ◽  
J. ELLSTEIN ◽  
C. A. McDEVITT

The formation of neuroma scar after trauma or neurorrhaphy is believed to be mediated by the response of collagen forming fibroblasts. In this study of twenty human neuromas, myofibroblasts were identified as a component of the scar. These cells occurred singly or as aggregates. There was a qualitative increase of myofibroblasts during the period from two to six months post-injury. From six months to one and one-half years post-injury, numbers and aggregations of myofibroblasts diminished, concurrent with collagen proliferation. Ultrastructural alcian blue staining and biochemical analyses revealed a glucosamine—glycosaminoglycan matrix within neuromas when compared to control nerves. Myofibroblasts appear to play a part in the pathobiology of human neuromas.


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