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International Journal of Clinical and... 2019The purpose of this study was to document histologic features of the herniated sublingual gland (SLG) and investigate the histologic correlation between herniated SLG...
OBJECTIVES/HYPOTHESIS
The purpose of this study was to document histologic features of the herniated sublingual gland (SLG) and investigate the histologic correlation between herniated SLG and plunging ranula.
METHODS
One hundred half-heads from 50 adult cadavers (21 females and 29 males) were included in this study. The presence of SLG herniation and the histologic features SLG were analyzed. The histologic features were analyzed according to the part: intraoral, junctional, and herniated parts. Hematoxylin and eosin (H&E), periodic acid Schiff reaction (PAS), and Alcian Blue (pH 2.5) staining were performed.
RESULTS
SLG herniation was found in 42 of 100 half-heads. Non-herniated SLG and the intraoral part of the herniated SLG were mainly composed of mucous acini and a few mixed acini. Junctional and herniated parts were mainly composed of serous acini and showed fatty change. PAS and Alcian blue staining showed that both acidic and neutral mucinous acini of junctional and herniated parts were decreased. However, there was no pseudo-epithelium at any site of herniation.
CONCLUSIONS
The histologic features of herniated SLG are different according the portions. The herniated part showed fatty degeneration and the remaining acini were mainly serous. We cannot confer any correlation between plunging ranula and the herniated part of SLGs.
PubMed: 31933831
DOI: No ID Found -
Regenerative Biomaterials 2022Autologous chondrocytes (C cells) are effective sources of cell therapy for engineering cartilage tissue to repair chondral defects, such as degenerative arthritis. The...
Autologous chondrocytes (C cells) are effective sources of cell therapy for engineering cartilage tissue to repair chondral defects, such as degenerative arthritis. The expansion of cells with C cell characteristics has become a major challenge due to inadequate donor sites and poor proliferation of mature C cells. The perichondrial progenitor cells (P cells) from the cambium layer of the perichondrium possessed significantly higher mesenchymal stem cell markers than C cells. In the transwell co-culture system, P cells increased the passaging capacity of C cells from P6 to P9, and the cell number increased 128 times. This system increased the percentage of Alcian blue-positive C cells from 40% in P6 to 62% in P9, contributing about 198 times more Alcian blue-positive C cells than the control group. C cells co-cultured with P cells also exhibited higher proliferation than C cells cultured with P cell-conditioned medium. Similar results were obtained in nude mice that were subcutaneously implanted with C cells, P cells or a mixture of the two cell types, in which the presence of both cells enhanced neocartilage formation . In aggregate, P cells enhanced the proliferation of C cells in a dose-dependent manner and prolonged the longevity of mature C cells for clinical applications.
PubMed: 35702349
DOI: 10.1093/rb/rbab078 -
Archives of Medical Research Jul 2021Bone marrow mesenchymal stem cells (BMSCs) are ideal seed cells for tissue engineering cartilage construction. However, the underlying mechanism of it has not been...
AIM
Bone marrow mesenchymal stem cells (BMSCs) are ideal seed cells for tissue engineering cartilage construction. However, the underlying mechanism of it has not been illuminate well. In this study, the effects of circATRNL1 (hsa_circ_0020093) on the differentiation of BMSCs into chondrocytes were investigated.
METHODS
The degrees of chondrogenic differentiation of BMSCs on day 0, 14 and 21 mediums were detected by Alcian blue staining. Expressions of cartilage differentiation related factors SOX9, COL2 and Aggrecan, and circATRNL1 in BMSCs under differentiation were determined by western blot and quantitative real-time polymerase chain reaction (qRT-PCR) as needed. circATRNL1 knockdown or overexpression was performed in BMSCs. Then the viability of BMSCs and cartilage differentiation related factors were separately investigated through MTT assay, qRT-PCR, and western blot. Target gene of circATRNL1 and binding site were predicted using starbase and validated it by dual luciferase reporter. The effect of circATRNL1 and its target gene on chondrogenic differentiation of BMSCs was assessed using Alcian blue staining further.
RESULTS
The degrees of chondrogenic differentiation of BMSCs were increased with time. Expressions of SOX9, COL2 and Aggrecan as well as circATRNL1 were enhanced during chondrogenic differentiation. Furthermore, overexpression of circATRNL1 enhanced BMSCs proliferation, SOX9, COL2 and Aggrecan expressions and the degree of chondrogenic differentiation of BMSCs. Further research showed that circATRNL1 targeted miR-338-3p. MiR-338-3p inhibited differentiation of BMSCs into cartilage but overexpression of circATRNL1 reversed it.
CONCLUSION
CircATRNL1 is beneficial to BMSCs differentiation into cartilage by regulating miR-338-3p, which may be a new mechanism of action in the treatment of cartilage repair.
Topics: Cell Differentiation; Cells, Cultured; Chondrogenesis; Humans; Mesenchymal Stem Cells; MicroRNAs; RNA, Circular
PubMed: 33610389
DOI: 10.1016/j.arcmed.2021.02.003 -
Poultry Science Jul 2021As the largest single bone, avian sterna are very different from those of mammals in terms of morphology and functions. Moreover, years of artificial selection in...
As the largest single bone, avian sterna are very different from those of mammals in terms of morphology and functions. Moreover, years of artificial selection in poultry led to incomplete sternal ossification at slaughter age, which may cause diseases, sternal injury, and restriction to breast muscle growth. However, in living birds, studies have rarely described the ossification pattern and underlying mechanisms of the sterna. Here, we examined the pattern (timeline, ossification centers, ossification directions, weekly changes of different parts, quantified differences in ossification degree among sexes and parts) and developmental changes (histological structure, gene expression) of postnatal duck sternal ossification. Direct observation and alcian blue and alizarin red staining of whole sterna samples revealed that, duck sterna mainly ossified during 5 to 9 wk old with five ossification centers. These centers and their ossification directions were different from and more complex than the previously studied birds. The weekly changes of sterna and the quantitative analysis of ossification-related traits showed that ossifications in the three parts of duck sterna (sternum body, keel, posterolateral processes) were mutually independent in space and time, meanwhile, the male duck sterna were more late-maturing than the female. The results of hematoxylin-eosin, alcian blue, and toluidine blue stainings and the expression levels of COL2A1, COL10A1, COL1A2, and CTSK together supported that, duck sternal ossification was highly similar to typical endochondral ossification. Furthermore, continuously high expression of MMP13 and SPARC and their significant (P < 0.05) co-expression with COL2A1, COL10A1, COL1A2, and CTSK suggested the importance of MMP13 and SPARC in duck sternal ossification. Taken together, our results may be helpful for the understanding of avian sternal ossification and the improvement of the performance and welfare of poultry from a new perspective.
Topics: Animals; Chickens; Ducks; Female; Gene Expression; Male; Osteogenesis; Sternum
PubMed: 34116350
DOI: 10.1016/j.psj.2021.101112 -
Contrast Media & Molecular Imaging 2022To address the question of determining the osteogenic differentiation of mesenchymal stem cells, the bone marrow studies were performed using probe microscopy. All...
To address the question of determining the osteogenic differentiation of mesenchymal stem cells, the bone marrow studies were performed using probe microscopy. All adherent bone marrow was used to isolate the bone marrow mesenchymal stem cells and expanded and purified in vitro. Its morphology under an inverted microscope was observed. We used Zuogui Pills to differentiate the separation methods. Alcian blue staining, modified calcium cobalt alkaline phosphatase staining, and neuron-specific enolase immunohistochemical staining were performed. The experimental results are shown below. The morphology of the isolated and purified cells was analyzed with an inverted microscope, and the isolated and purified cells were analyzed with Zuogui Pill. Alcian blue staining, modified calcium cobalt alkaline phosphatase staining, and neuron-specific enolase immunohistochemical staining confirmed that the cells differentiated into cartilage and osteoblasts, and the cell structure and morphology were similar to those of the bone marrow mesenchymal stem cells. The results showed that the adherent mode of cells obtained from the whole bone marrow was the rat bone marrow mesenchymal stem cells, and the Zuogui Pills could induce multidirectional differences in the bone marrow mesenchymal stem cells.
Topics: Alcian Blue; Alkaline Phosphatase; Animals; Bone Marrow; Bone Marrow Cells; Calcium; Cells, Cultured; Cobalt; Mesenchymal Stem Cells; Microscopy, Scanning Probe; Osteogenesis; Phosphopyruvate Hydratase; Rats
PubMed: 35854771
DOI: 10.1155/2022/6483087 -
Journal of Acupuncture and Meridian... Jun 2020The primo vascular system (PVS) has been difficult to detect due to its small diameter and translucent features of the threadlike network. Thus, contrast-enhancing dyes...
BACKGROUND
The primo vascular system (PVS) has been difficult to detect due to its small diameter and translucent features of the threadlike network. Thus, contrast-enhancing dyes including Alcian blue, Trypan blue and Janus green B had to be used for finding and taking out PVS from rat and mouse.
OBJECTIVE
Generation of monoclonal antibodies (mAbs) against PVS of rat was intended to use as a detector for PVS and a biological tool for functional study of PVS.
MATERIALS AND METHODS
Primo vessel (PV) and Primo node (PN) were isolated from organ surfaces of rat and then their proteins were isolated and injected into mouse as an immunogen. The classical traditional method was applied for production of mAbs against PVS. The various techniques, such as cell fusion, screening of hybridoma, ELISA, Western blotting (WB), immunofluorescence microscopy (IF), and limiting dilution, were used to generate mAbs against PVS.
RESULTS
Among 16 mAbs generated, 4 representative mAbs were characterized with their specificities in ELISA, WB, and IF. α-rPVS-m1-1 and α-rPVS-m4-6 had strong binding affinities to PVS in both ELISA and WB but did not show specificities in IF at all. On the contrary, α-rPVS-m3-2 and α-rPVS-m3-4 almost did not respond in WB but had strong binding affinities in ELISA and specificities in IF. Two mAbs stained predominantly at extra cellular matrix and cell membrane of PVS of rat in IF, and they were able to discriminate PVS from blood vessel (BV) and lymphatic vessel (LV).
CONCLUSIONS
4 representative mAbs against PVS of rat were characterized by ELISA, WB, and IF. α-rPVS-m3-2 and α-rPVS-m3-4, which had strong specificities in IF, can be used as a tool in discriminating PVS from other similar tissues and in elucidate biological function of PVS.
Topics: Alcian Blue; Animals; Antibodies, Monoclonal; Lymphatic Vessels; Male; Meridians; Mice; Rats; Rats, Sprague-Dawley; Staining and Labeling
PubMed: 32437979
DOI: 10.1016/j.jams.2020.05.001 -
The American Journal of Pathology May 1982Dwarfism in the Norwegian Elkhound occurred as a result of a generalized disturbance in endochondral ossification. Radiographic changes included flaring and increased...
Dwarfism in the Norwegian Elkhound occurred as a result of a generalized disturbance in endochondral ossification. Radiographic changes included flaring and increased width of the distal metaphyses of the radius and ulna, delayed ossification of the cuboid bones of the carpus, and reduction in length of the vertebral bodies. The zone of chondrocyte proliferation was decreased in width and contained areas of abnormal cell column formation alternated with wide areas of matrix. Chondrocytes in all zones contained one or more inclusions bounded by a smooth discontinuous membrane. The material within the inclusions appeared homogeneous and stained blue-green with Movat's pentachrome and deep blue with alcian blue-periodic acid-Schiff at pH 1.0 and 2.6. The distribution of ruthenium red granules in the matrix frequently revealed poor differentiation into territorial and interterritorial zones. Twenty-four-hour urine samples were negative for glucose, and the glycosaminoglycan excretion pattern was normal.
Topics: Age Factors; Amino Acids; Animals; Calcium; Cervical Vertebrae; Dog Diseases; Dogs; Dwarfism; Female; Glycosuria; Hematocrit; Male; Radiography; Radius; Ulna
PubMed: 7081383
DOI: No ID Found -
Ocular Oncology and Pathology Apr 2018To investigate the source of fibrous astrocytes and neuroblasts in a small ciliary body medulloepithelioma appearing as a leukocoria in a 3-week-old baby girl.
PURPOSE
To investigate the source of fibrous astrocytes and neuroblasts in a small ciliary body medulloepithelioma appearing as a leukocoria in a 3-week-old baby girl.
METHODS
Histopathologic and immunohistochemical studies included Alcian blue, periodic acid-Schiff, and antisera for the detection of S100 protein, CD99, glial fibrillary acidic protein (GFAP), CRX, NeuN, neurofilaments, synaptophysin, desmin, and myogenin.
RESULTS
A small, nonteratoid ciliary body medulloepithelioma with collections of Alcian blue+ mucoplysaccharides was present in the enucleated globe. The retinal mass displayed multilaminar dysplastic rosettes that were CRX+, NeuN-, and synaptophysin-. Intraretinal neurofilaments and scattered NeuN+ neurocytes were also identified. At the base of the retinal mass ribbons and pseudopapillae of CRX+, NeuN- medullary epithelium were found. The latter developed from an S100+ and weakly CD99+ monolayer of premedullary epithelium. GFAP+ fibrous astrocytes and NeuN- neuroblasts streamed from the medullary epithelium.
CONCLUSIONS
A multilaminar medullary epithelium and a precursor monolayer of premedullary epithelium were both identified. Neuroblasts and fibrous astrocytes were determined to arise separately from the medullary epithelium. The early stage of tumorigenesis afforded by a small tumor provided the opportunity to discover morphologic and immunohistochemical evidence for these differentiations.
PubMed: 29765950
DOI: 10.1159/000481287 -
Journal of Functional Biomaterials Apr 2019Accurate determination of the amount of glycosaminoglycans (GAGs) in a complex mixture of extracellular matrix (ECM) is important for tissue morphogenesis and...
Accurate determination of the amount of glycosaminoglycans (GAGs) in a complex mixture of extracellular matrix (ECM) is important for tissue morphogenesis and homeostasis. The aim of the present study was to investigate an accurate, simple and sensitive alcian blue (AB) method for quantifying heparin in biological samples. A method for analyzing heparin was developed and parameters such as volume, precipitation time, solvent component, and solubility time were evaluated. The AB dye and heparin samples were allowed to react at 4 ℃ for 24 h. The heparin-AB complex was dissolved in 25 N NaOH and 2-Aminoethanol (1:24 /). The optical density of the solution was analyzed by UV-Vis spectrometry at 620 nm. The modified AB method was validated in accordance with U.S. Food and Drug Administration guidelines. The limit of detection was found to be 2.95 µg/mL. Intraday and interday precision ranged between 2.14-4.83% and 3.16-7.02% (n = 9), respectively. Overall recovery for three concentration levels varied between 97 ± 3.5%, confirming good accuracy. In addition, this study has discovered the interdisciplinary nature of protein detection using the AB method. The basis for this investigation was that the fibrous protein inhibits heparin-AB complex whereas globular protein does not. Further, we measured the content of sulfated GAGs (sGAGs; expressed as heparin equivalent) in the ECM of decellularized porcine liver. In conclusion, the AB method may be used for the quantitative analysis of heparin in ECM scaffolds for tissue engineering applications.
PubMed: 31052349
DOI: 10.3390/jfb10020019 -
Current Biology : CB Jun 2022The kinetochore links chromosomes to spindle microtubules to drive chromosome segregation at cell division. While we know nearly all mammalian kinetochore proteins, how...
The kinetochore links chromosomes to spindle microtubules to drive chromosome segregation at cell division. While we know nearly all mammalian kinetochore proteins, how these give rise to the strong yet dynamic microtubule attachments required for function remains poorly understood. Here, we focus on the Astrin-SKAP complex, which localizes to bioriented kinetochores and is essential for chromosome segregation but whose mechanical role is unclear. Live imaging reveals that SKAP depletion dampens the movement and decreases the coordination of metaphase sister kinetochores and increases the tension between them. Using laser ablation to isolate kinetochores bound to polymerizing versus depolymerizing microtubules, we show that without SKAP, kinetochores move slower on both polymerizing and depolymerizing microtubules and that more force is needed to rescue microtubules to polymerize. Thus, in contrast to the previously described kinetochore proteins that increase the grip on microtubules under force, Astrin-SKAP reduces the grip, increasing attachment dynamics and force responsiveness and reducing friction. Together, our findings suggest a model where the Astrin-SKAP complex effectively "lubricates" correct, bioriented attachments to help preserve them.
Topics: Alcian Blue; Animals; Cell Cycle Proteins; Chromosome Segregation; Friction; Kinetochores; Mammals; Microtubule-Associated Proteins; Microtubules; Mitosis; Phenazines; Phenothiazines; Resorcinols; Spindle Apparatus
PubMed: 35580605
DOI: 10.1016/j.cub.2022.04.061