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  • 1
    ISSN: 1432-0827
    Keywords: Periodontal ligament fibroblast ; Mineralized nodule ; Ultrastructure
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Physics
    Notes: Summary The purposes of this study were to determine whether periodontal ligament (PDL) cells are capable of producing mineralized nodules in vitro and to analyze ultrastructural features of the nodules. Rat PDL cells were obtained from coagulum in the socket at 2 days after tooth extraction and cultured at confluence in standard medium containing Dulbecco's Modified Eagle's Medium supplemented with 10% FBS and antibiotics. To test mineralized nodule formation, cells were further cultured for an additional 3 weeks in the standard medium containing (1) ascorbic acid (50 μg/ml) and sodium β-glycerophosphate (10 mM), (2) ascorbic acid, sodium β-glycerophosphate, and dexamethasone (5 μM), or (3) ascorbic acid alone. Cells were then fixed in 2.5% glutaraldehyde, postfixed in 1% OsO4, and prepared for light and electron microscopy. Threedimensional nodules containing mineralized matrices were formed only when the cells were cultured in the presence of ascorbic acid and dexamethasone. They were composed of multilayered fibroblasts (up to 13 layers), and highly organized collagen fibrils with 64 nm cross-banding patterns between the cell layers. The fibroblasts in the nodules exhibited an elongated shape with a high degree of cytoplasmic polarity throughout the nodule, and have the morphological features of PDL fibroblasts as seen in vivo. Mineral deposition with needle-like crystals was initiated on collagen fibrils located in intercellular spaces of the upper cell layers and became increasingly heavier towards the bottom half of the nodules. X-ray microanalysis and electron diffraction analysis confirmed that mineral deposition contained calcium and phosphate in the form of immature hydroxyapatite. These nodules contained neither osteoblasts nor osteocytes, and have their own morphological organization and characteristics which differ from those formed by bone cells in culture. Therefore, these data suggest that PDL cells are capable of forming mineralized tissue in vitro with the morphological characteristics different from bone mineralized nodules.
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Journal of periodontal research 34 (1999), S. 0 
    ISSN: 1600-0765
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Transforming growth factor-betas (TGF-βs) and bone morphogenetic proteins (BMPs), members of a TGF-β superfamily, are known to play an important role in osteogenic cell differentiation and consequently bone formation. We have reported previously that periodontal ligament (PDL) cells differentiate and form mineralized nodules when cultured in the presence of dexamethasone (Dex), β-glycerophosphate (GP) and ascorbic acid (AA). To understand the roles of TGF-β isoforms (TGF-β1, 2 and 3) and TGF-β type 1 receptors (activin receptor-like kinase (ALK)-2, -3, -5 and -6) in PDL cell differentiation, their expression was investigated using Northern blot analysis. Rat PDL cells, derived from coagulum in the tooth socket, were cultured in the presence of Dex (5 μM), GP (10 mM) and AA (50 μg/ml) for up to 21 d. Total RNA was isolated from PDL cells after 0, 7, 14 and 21 d and used for Northern blot analysis of mRNAs for matrix proteins, TGF-β isoforms and their receptors using 32P-labeled cDNAs as probes. Four stages showing distinct morphological characteristics and matrix expression during development of mineralized nodules were identified. Type I collagen (Col I) and SPARC (secreted protein. acidic and rich in cysleine) mRNAs were expressed at the confluent stage, but decreased during the mineralization stage. Osteopontin (OPN) and alkaline phosphatase (ALP) transcripts were initially observed at multilayer stage, while bone sialoprotein (BSP) and osteocalcin (OC) at the nodule stage and all 4 were expressed thereafter. TGF-β1 mRNA expression increased with the progression of PDL cell differentiation, while a relatively high level of TGF-β3 transcript decreased slightly during their differentiation. TGF-β2 mRNA was not expressed. The expression of TGFβ-RI mRNA decreased, whereas that of TGFβ-RIII increased dramatically with PDL cell differentiation. TGFβ-RII gene activities remained high throughout all stages. ALK-2, ALK-3 and ALK-6 mRNA expression increased with the progression of PDL cell differentiation, suggesting that these receptors may play important roles in Dex-induced PDL cell differentiation and mineralized nodule formation.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1546-1718
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Medicine
    Notes: [Auszug] Neurofibrillary tangles (NFT) composed of the microtubule-associated protein tau are prominent in Alzheimer disease (AD), Pick disease, progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD). Mutations in the gene (Mtapt) encoding tau protein cause frontotemporal dementia ...
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Graefe's archive for clinical and experimental ophthalmology 226 (1988), S. 559-566 
    ISSN: 1435-702X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The ultrastructure and permeability of retinal arterioles, venules, and capillaries located near the optic disc of stroke-prone spontaneously hypertensive rats (SHRSP) and normotensive Wistar-Kyoto rats were studied using electron microscopy and tracer cytochemistry. A variety of structural changes were observed in arterioles of SHRSP. They include (1) narrowing of the lumen due to smooth muscle hyperplasia and/or fragmentation and thickening of the basal lamina, (2) fusiform aneurysms containing degenerated smooth muscle cells and reduplicated basal lamina, and (3) the presence of microfilament bundles under the luminal surface of the endothelium. In addition, the wall of venules was thickened due to accumulation of basal lamina material. Many capillary pericytes were also degenerated. Retinal vessels of age-matched normotensive rats did not show such changes. In SHRSP, after injection of peroxidase, extravasation of tracer was seen occasionally in retinal capillaries and in the central retinal vein at the optic nerve head. No changes in vascular permeability were observed in the normotensive rats.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Graefe's archive for clinical and experimental ophthalmology 229 (1991), S. 457-463 
    ISSN: 1435-702X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Experimental autoimmune uveoretinitis was induced in Lewis rats by footpad injection of purified, bovine retinal soluble antigen (S-antigen) with complete Freund's adjuvant. Control animals received adjuvant only. At the peak of ocular inflammation the animals received an intravenous injection of the tracer horseradish peroxidase, and the eyes were removed and processed for peroxidase cytochemistry. A breakdown of the blood-retinal barrier occurred at the level of the retinal venules. Peroxidase reaction product was demonstrable in the interendothelial tight junctions and in the perivascular spaces. Heavy infiltration of inflammatory cells was also found in and around the venular wall. The endothelial cells of venules and capillaries were hypertrophied and showed swollen endoplasmic reticulum and Golgi apparatus as well as increased numbers of ribosomes and cytoplasmic fibrils; pericytes and smooth-muscle cells showed a similar synthetic phenotype and the basal lamina of these cells was thickened.
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  • 6
    ISSN: 0003-276X
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Occurrence of epidermal growth factor (EGF)-binding sites during differentiation of cementoblasts and periodontal ligament (PDL) fibroblasts was investigated using radioautography after I. V. injection of 125I-EGF to 14-day-old rats. During differentiation of cementoblasts, a very low level of EGF-binding sites was present on the mesenchymal cells in dental follicle proper, precementoblasts, and cementoblasts. On the other hand, during differentiation of PDL fibroblasts, numerous EGF-binding sites were observed on the undifferentiated paravascular cells and on the perifollicular mesenchymes representing the major source of PDL fibroblast precursor cells. Also heavy labeling was observed throughout their differentiation to PDL fibroblasts, as well as during full synthetic activity as mature cells. Quantitative analysis of the light microscopic radioautographs revealed that these cells demonstrated approximately 4 grains per 100μm2 of cell area. These results suggest that EGF plays an important role in differentiation of PDL fibroblasts, but not in that of cementoblasts. Furthermore, the well-known in vivo effect of EGF in producing precocious eruption of teeth may be a consequence of a more extensive effect of EGF throughout differentiation of PDL fibroblasts as well as during full synthetic activity as mature cells.
    Additional Material: 10 Ill.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    The @Anatomical Record 240 (1994), S. 492-506 
    ISSN: 0003-276X
    Keywords: PDL fibroblast ; Cell differentiation ; Osteoblast ; Socket healing ; Radioautography ; Bromodeoxyuridine ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Background: The entire socket after tooth extraction is filled with new bone formed by osteoblasts (Obs), but the origin of these Obs remains unknown. Thus, the proliferation and migration of paravascular and endosteal fibroblastic cells and periodontal ligament (PDL) fibroblasts (Fbs) and their differentiation into Obs during socket healing after extraction of the first maxillary molars of the rat were investigated.Methods: The proliferative activity and migration of these cells in the sockets after tooth extraction were studied using radioautography and immunohistochemistry after injection of 3H-thymidine and 5-bromo-2′-deoxy-uridine (BrdU), respectively. Their morphological changes during differentiation was investigated by transmission electron microscopy.Results: One day after tooth extraction, PDL Fbs were the major cell type in the PDL remnant of the socket. Proliferation was low (labeling index (LI) = approximately 2%) until 16 h after tooth extraction but dramatically increased to a maximum level 1 day postextraction (LI = 23%). Between 1 and 2 days, numerous PDL Fbs in the PDL remnant actively migrated into the coagulum and continued to proliferate. On the basis of the high proliferative activity and small number of cellular organelles responsible for procollagen synthesis, these cells appear immature. At 3 days, Fbs contained more cellular organelles and deposited more collagen fibers as they replaced the coagulum with dense connective tissue and the LI declined. At 4 and 5 days, some of the Fbs began to differentiate into Obs, and the proliferation of Fbs dramatically decreased to baseline values. The migration of PDL Fbs and their differentiation into Obs were investigated by labeling with 3H-thymidine or BrdU 1 day after tooth extraction. Heavily labeled Fbs were observed in the PDL remnant at 1 day, in the coagulum at 2 days, and in the dense connective tissue at 3 days. Labeled Obs associated with new bone were seen 4 days after injection. Endosteal and paravascular Fbs also proliferated, but at a lower level and at later time periods than the PDL Fbs. Surprisingly, endosteal and paravascular Fbs contributed only a small population of Fbs to socket healing.Conclusions: These results indicate that PDL Fbs after tooth extraction actively proliferate, migrate into the coagulum, form dense connective tissue, and differentiate into Obs which form new bone during socket healing. © 1994 Wiley-Liss, Inc.
    Additional Material: 10 Ill.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 198 (1979), S. 119-127 
    ISSN: 1432-0878
    Keywords: Male hamster ; Harderian gland ; Castration ; Sexual dimorphism ; Porphyrin ; Testosterone ; Tubular clusters ; Membranous structures ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary A sexual dimorphism of the hamster Harderian gland at the ultrastructural level has been reported. The effect of testosterone on the fine structure of the gland from castrated male golden hamsters is reported here. Harderian glands from the following three groups of animals were examined at regular intervals up to 60 days after castration: (1) castrated; (2) castratedsham-injected, receiving 0.1 ml sesame oil per day; (3) castrated-testosterone injected, receiving 2mg testosterone propionate in 0.1 ml sesame oil per day. In groups 1 and 2, clusters of cylindrical tubules, typical of the male gland, decreased in number and disappeared almost completely 2 weeks after castration. Membranous structures, typical of the female gland, prevailed in these two groups throughout the remaining period of experiment. On the other hand, these changes were prevented in the group of castrated animals maintained on testosterone propionate. It is concluded that castration modified the ultrastructure of the male hamster Harderian gland toward the female type and that daily administration of testosterone propionate prevented this change.
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 245 (1986), S. 431-437 
    ISSN: 1432-0878
    Keywords: Retina ; Arteriole ; Venule ; Tannic acid ; Peroxidase ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The surface-associated vesicles in retinal arterioles and venules were studied after fixation in glutaraldehyde-tannic acid or after intravitreal injection of peroxidase or lactoperoxidase. The vesicles were concentrated along the abluminal (basal) surface of the endothelial cells and along the plasma membranes of smooth muscle cells in arterioles and of pericytes in post-capillary venules. They were rarely encountered in the deeper regions of these cells. In perpendicular sections through the cell surface the majority of vesicles were in continuity with the plasma membrane whereas in tangential sections, they appeared to lie “free” in the cytoplasm. All such vesicles were labeled after exposure to tannic acid or to the heme-proteins. Peroxidase-reaction product was never seen in the lumen of the vessels. These observations suggest that the surface vesicles in endothelial cells, smooth muscle cells and pericytes are invaginations of the plasma membrane and are thus not involved in the transcytosis or endocytosis of proteins. The vesicles in the latter two cell types may be involved in some aspect of contractility rather than pinocytosis.
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