Alcian Blue Staining Technique Research Articles

Effects of extracellular vesicles from adipose-derived stem cells on human keloid fibroblasts via the SOCS1/JAK2/STAT3 pathway.

Keloid represents a benign skin tumor with many cancer-like features. Extracellular vesicles (EVs) derived from human adipose-derived stem cells (hADSCs) play a role in cell migration of multiple diseases. This study aimed to investigate the impact of hADSC-EVs on human keloid fibroblasts (HKFs). hADSCs were cultured to the 3rd generation, and subsequently assessed for their osteogenic, adipogenic, and chondrogenic differentiative abilities using flow cytometry, alizarin red, oil red O, and alcian blue staining techniques. hADSC-EVs were isolated through ultracentrifugation and subsequently identified. HKFs at the 3rd generation were subjected to treatment with hADSC-EVs to observe their endocytosis of EVs by immunofluorescence. CCK-8, wound healing, and Transwell assays were performed to test HKF proliferation and migration. The levels of autophagy proteins, collagens, and Janus kinase 2 (JAK2) and Signal Transducer and Activator of Transcription 3 (STAT3) were determined through Western blot analysis. Suppressor of cytokine signaling 1 (SOCS1) expression was determined by RT-qPCR and Western blot. hADSC-EVs were successfully isolated from hADSCs. PKH67-labeled hADSC-EVs were observed to be endocytosed by HKFs, resulting the inhibition of HKF proliferation, migration, as well as a reduction in collagen deposition. hADSC-EVs carried SOCS1 into HKFs to suppress HKF autophagy. SOCS1 downregulation in hADSC-EVs partially nullified the inhibitory effect of hADSC-EVs on HKFs. hADSC-EV-carried SOCS1 inhibited the activation of the JAK2/STAT3 pathway. JAK2/STAT3 pathway activation partially abrogated the suppression of hADSC-EVs on the proliferation, migration, and collagen deposition of HKF. hADSC-EVs carried SOCS1 into HKFs and suppressed HKF autophagy, proliferation, migration, and collagen deposition by inactivating the JAK2/STAT3 pathway.

Development and Characterization of Decellularized Lung Extracellular Matrix Hydrogels.

The use of extracellular matrix (ECM)-derived hydrogels in tissue engineering has become increasingly popular, as they can mimic cells' natural environment in vitro. However, maintaining the native biochemical content of the ECM, achieving mechanical stability, and comprehending the impact of the decellularization process on the mechanical properties of the ECM hydrogels are challenging. Here, a pipeline for decellularization of bovine lung tissue using two different protocols, downstream characterization of the effectiveness of decellularization, fabrication of reconstituted decellularized lung ECM hydrogels and assessment of their mechanical and cytocompatibility properties were described. Decellularization of the bovine lung was pursued using a physical (freeze-thaw cycles) or chemical (detergent-based) method. Hematoxylin and Eosin staining was performed to validate the decellularization and retention of major ECM components. For the evaluation of residual collagen and sulfated glycosaminoglycan (sGAG) content within the decellularized samples, Sirius red and Alcian blue staining techniques were employed, respectively. Mechanical properties of the decellularized lung ECM hydrogels were characterized by oscillatory rheology. The results suggest that decellularized bovine lung hydrogels can provide a reliable organotypicalternative to commercial ECM products by retaining most native ECM components. Furthermore, these findings reveal that the decellularization method of choice significantly affects gelation kinetics as well as the stiffness and viscoelastic properties of resulting hydrogels.

Modulation of Small-Intestine Morphology in Mice by a Novel Supplement Containing Silybum marianum, Yeast β-Glucan, Prebiotics, and Minerals

AbstractSilymarin, derived from Silybum marianum, has recently demonstrated its potential to improve health in conditions such as obesity and metabolic disturbances. Understanding the impact of nutraceuticals on intestinal morphology is crucial for developing supplements that promote a higher quality of life. Therefore, this study aimed to investigate the effects of nutraceutical supplementation with silymarin on the morphology of the small intestine. Sixty-day-old adult male C57BL/6 mice were divided into two groups: one receiving a standard chow (control) and the other receiving a novel silymarin supplement (experimental). Following the experimental period, the animals were euthanized, and fragments of the small intestine were collected for histochemical analysis using Masson's trichrome and periodic acid-Schiff with Alcian blue staining techniques. Our results revealed an increase in the number of villi per analyzed field in the experimental group, accompanied by a decrease in basic mucin, crypt depth, mucosal thickness, and villus spacing. In conclusion, this novel nutraceutical supplementation may play a crucial role in modulating small intestine morphology and enhancing absorption capacity.

Study the Thorax Ossification Centers in Partridge Embryos by Alizarin Red and Alcian Blue Double Staining Techniques

The current study was aimed to determine the onset of vertebral ossification, ribs, and sternum, and to evaluate ossification centers using the alizarin red and alcian blue staining techniques in the partridge embryos. One-hundred fertilized eggs of the partridge were placed in a hatching machine. From day 8 of incubation, three egg samples were randomly selected daily for staining until hatching. After staining, ossification centers in various parts of thorax were evaluated by a stereomicroscope. Data from examination of different thoracic parts of the partridge embryos indicated that cartilaginous thoracic vertebrae was were formed in the embryo on the 8th day, but the ribs and sternum were not visible. Cartilaginous ribs were formed on the 9th day of embryo. A cartilaginous sternum began to form on the 10thday of embryo. On the 13th day, cartilaginous uncinate processes were observed at the caudal border of embryonic ribs. From the 16th day, the first ossification center was detected in the body of ribs. The embryonic development process was progressive on the 17th and 18th days. The first ossification centers in the embryo of the 22nd and 23rd days were detectable in the cranio-lateral process of sternum and in the uncinate process of the first thoracic rib, respectively. On the 25th day, the cranio-medial process of sternum was the first ossification center in the embryos. The emergence of ossification centers in different parts of thorax and the developmental process were not observed at the pre-hatching time.

Histochemistry of Equine Damaged Tendons, Ligaments and Articular Cartilage

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.

Age determination with alcian blue staining technique for Lagocephalus sceleratus in Mersin Bay, Turkey, northeastern Mediterranean

In this study, alcian blue staining technique was used to enhance the visibility of the band on vertebrae of Lagocephalus sceleratus from Mersin Bay, Turkey, Northeastern Mediterranean. A total of 77 L. sceleratus samples were captured by commercial purse seine nets at depths between 24 and 50 m from Mersin Bay (Turkey). The total length ranged from 8.9 to 78.4 cm and body weight varied between 7.59-4750 g for L. sceleratus. Age determinations were carried out using whole vertebrae. A total of 4 vertebrae from each specimen were read by two independent readers. All images were taken using a Leica S8APO brand microscope with a high resolution Leica Application suit (Las V4.8) software. Vertebral images were then enhanced using Adobe Photoshop CS2 to improve sharpness and clarity. Of all the 77 staining vertebrae were readable and aged by both readers, resulting in an IAPE of 7.8 %. The maximum age was found as 10+ years old.

Production of transparent exopolymer particles (TEP) by the eastern oyster Crassostrea virginica

MEPS Marine Ecology Progress Series Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsTheme Sections MEPS 288:141-149 (2005) - doi:10.3354/meps288141 Production of transparent exopolymer particles (TEP) by the eastern oyster Crassostrea virginica Michael P. McKee1,*, J. Evan Ward1, Bruce A. MacDonald2, Bridget A. Holohan1 1Department of Marine Sciences, University of Connecticut, 1080 Shennecossett Road, Groton, Connecticut 06340, USA2Department of Biology, University of New Brunswick, Saint John, PO Box 5050, New Brunswick E2L 4L5, Canada *Email: mckee@uconn.edu ABSTRACT: Transparent exopolymer particles (TEP) play an important role in aggregation, phytoplankton bloom dynamics and the fate of organic carbon. Various types of plankton are known to release exopolymers that contribute to the TEP pool. Little is known, however, about other potential sources of TEP and TEP precursors in the oceans, especially in nearshore waters. Suspension-feeding bivalves can form dense assemblages in coastal waters and can process large volumes of water during feeding activities (e.g. 5 l h–1 g–1 dry wt). They secrete mucus to maintain clean body surfaces and to aid in capturing and transporting food material. We hypothesized that mucopolysaccharides, which are released from bivalves in dissolved or particulate form, contribute to the TEP pool. To test this hypothesis, laboratory experiments were carried out to measure the concentration of TEP and DOC in chambers with actively feeding oysters Crassostrea virginica over a 9 h period. Field experiments were conducted to measure TEP and DOC production by oysters in situ, using clear Plexiglas chambers on an intertidal sand-flat in New Brunswick, Canada. TEP concentrations were determined with an Alcian blue staining technique and spectrophotometric detection. DOC concentrations were determined using high-temperature catalytic oxidation (HTCO) with a platinum catalyst. Laboratory results indicated that oysters significantly enhanced TEP and DOC concentrations compared to control levels over 9 h. Likewise, field experiments demonstrated elevated TEP concentrations in experimental chambers compared to control treatments. Our results support the hypothesis that oysters contribute to the production of TEP in coastal marine ecosystems. KEY WORDS: Transparent exopolymer particles · TEP · Mucopolysaccharides · Oysters · Dissolved organic carbon · DOC · Aggregation Full text in pdf format PreviousNextExport citation RSS - Facebook - Tweet - linkedIn Cited by Published in MEPS Vol. 288. Online publication date: March 10, 2005 Print ISSN: 0171-8630; Online ISSN: 1616-1599 Copyright © 2005 Inter-Research.

Hyaluronic acid metabolism in keloid fibroblasts

Hyaluronic acid (HA), an important component of the tissue extracellular matrix, is a ubiquitous glycosaminoglycan (GAG) that forms a pericellular coat on the surface of cells. It has been speculated that this pericellular HA boundary may localize cytokines, such as transforming growth factor-β1, which is known to stimulate collagen production. The purpose of this study was to examine the role of HA and its cell surface receptor (CD44), an active participant in HA degradation, as they relate to keloid formation. Dermal excisions from both normal patients (n = 13) and keloid patients (n = 13) were analyzed for HA content using an alcian blue staining technique. Fibroblast cell cultures were used to quantitate HA synthesis and CD44 receptor density. Histological analyses showed a greater HA content in keloid tissue compared with normal dermal tissue. In agreement with this observation, keloid fibroblasts were found to synthesize significantly more HA than normal dermal fibroblasts (2469 ± 483 cpm versus 1122 ± 256 cpm, P = .02). Treatment of keloid fibroblasts with triamcinolone acetonide reduced the level of HA synthesis to that of normal fibroblasts (1560 ± 477 cpm versus 1293 ± 264 cpm, P = .6). However, there was no significant difference in HA receptor density on keloid cells compared with normal skin fibroblasts. Therefore, the increased HA deposits found in keloids are attributable to increased synthesis rather than to decreased degradation mediated by the CD44 receptor.

Maturation of glomerular size distribution profiles in domestic fowl (Gallus gallus)

Previous histological evaluations of chick kidneys indicated nephrons continue to develop from embryonic foci for up to 6 weeks after hatching. The present study was conducted using an in vivo alcian blue staining technique to quantify posthatch changes in glomerular numbers and sizes in female domestic fowl at 1, 3, 5, 9, 12, 21, and 30 weeks of age. Changes in glomerular size distributions reflect changes in the heterogeneous nephron populations of avian kidneys. Foci of embryonic tissue were observed at the periphery of renal lobules up to 12 weeks of age. Glomerular numbers increased from 69,800/kidney at 1 week to 586,000/kidney at 12 weeks, with no further significant increase up to 30 weeks (599,000/kidney). The increase in glomerular number per gram kidney weight remained constant as kidney mass increased up to 12 weeks of age, after which the number of glomeruli per gram kidney weight declined significantly as kidney size increased without further addition of new nephrons. Glomerular size distribution profiles were constructed using eleven circumference categories. The peak number of glomeruli fell within the 0.11-0.14 mm category at 1 and 3 weeks; within the 0.15-0.18 mm category at 5, 9, and 12 weeks; and within the 0.19-0.22 mm category at 21 and 30 weeks. One and 3-week-old chicks had no glomeruli within the largest (greater than or equal to 0.35 mm circumference) size categories, and 9-12-week-old birds had significantly fewer glomeruli in these categories than 21-30-week-old birds. These results demonstrate that posthatch renal maturation in domestic fowl involves the ongoing formation of new nephrons up to 12 weeks of age, with subsequent kidney growth (12-30 weeks of age) accomplished by enlargement of existing nephrons (nephron hypertrophy). The cumulative evidence indicates that nephrons destined to develop loops of Henle (mammalian-type) develop first, with shorter (reptilian-type) nephrons developing later as the kidneys enlarge.

Effect of orally administered food-grade carrageenans on antibody-mediated and cell-mediated immunity in the inbred rat

Experiments were performed to investigate the immunological consequences associated with the persorption of poorly degradable carregeenans from the diet. Using an inbred strain of rat it was demonstrated histochemically, by the carrageenan-specific Alcian blue staining technique, that small quantities of food-grade carrageenans given at 0.5% in drinking-water for 90 days could penetrate the intestinal barrier of adult animals. This apparently occurred via an intact mucosa in the absence of inflammatory or pathological lesions. The carrageenan was demonstrated in macrophage-like cells present within the villi and lamina propria of the small intestine. The oral administration of κ, λ or ı food-grade carrageenans did not affect local (biliary) or systemic antibody responses to gut commensal microorganisms, or to orally-administered sheep erythrocytes. However, when sheep red blood cells were administered parenterally the ensuing anti-sheep red blood cell haemagglutinating antibody response was temporarily suppressed in carrageenan-fed rats. λ-Carrageenan and ı-carrageenan both significantly ( P ⩽ 0.01 and P ⩽ 0.05, respectively) reduced the mid-phase (14–28 days) haemagglutinin response; κ-carrageenan (L100) was less effective but caused significant depression at day 21 ( P ⩽ 0.01). Individual responses were, however, within the control range 35 days after sheep erythrocyte administration, thus indicating the temporary nature of this effect. Although carrageenan administration depressed the anti-sheep erythrocyte antibody response, it did not affect T-cell immune competence as measured by the popliteal lymph node assay for graft-versus-host reactivity.

Histochemical studies on Echinorhynchus gadi (Acanthocephala)

The distribution of glycogen, polysaccharide, mucopolysaccharide, lipid and nucleic acid has been studied in Echinorhynchus gadi(Acanthocephala). The results were summarized as follows: 1. Glycogen and polysaccharide was demonstrated by Bauer PAS reaction technique and was found in fertilization membrane in ovum, central nuclear mass in acanthor and lemnisci, hypodermis in cystacanth. 2. Mucopolysaccharide was demonstrated by Mowry alcian blue staining technique and was found in outer membrane, fibrillar coat, fertilization membrane and inner membrane in acanthocephalan ova. 3. Lipid was demonstrated by Smith Nile blue stain and Lison Sudan black B staining technique and was found roughly parallel to that of polysaccharide. 4. Nucleic acid was demonstrated by Rosenbeck Feulgen reaction, Taft methylgreen-pyronin stain and Diengdoh acridine orange staining technique and found in central nuclear mass in acanthor, also, was found in lemnisci, proboscis and hypodermis in cystacanth.

Dermal glands and mucin in the moulting cycle of Rhodnius prolixus Stål.

The role of the dermal glands in the moulting cycle of Rhodnius has been reinvestigated by means of the alcian blue staining technique for mucin. Mucin is secreted by the majority of the dermal glands from the eleventh day after feeding until the time of moulting. The gland sacs become distended with mucin immediately prior to moulting, and this mucin is discharged and forms and inner lubricating layer on the exuvia during ecdysis.