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  • Immunfluorescenz  (3)
  • IN-VIVO  (2)
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  • 1
    Keywords: CANCER ; CELLS ; EXPRESSION ; IN-VITRO ; tumor ; CLINICAL-TRIAL ; human ; IN-VIVO ; MODEL ; liver ; PROTEIN ; PROTEINS ; DRUG ; TISSUE ; TUMORS ; MICE ; PATIENT ; SERA ; fibroblasts ; treatment ; MOUSE ; EFFICACY ; drug delivery ; CONJUGATE ; PHARMACOKINETICS ; BEARING RATS ; COLLAGEN-INDUCED ARTHRITIS ; FIBROBLAST-LIKE SYNOVIOCYTES ; LOW- DOSE METHOTREXATE ; SYNOVIAL FIBROBLASTS
    Abstract: We reported recently that albumin is a suitable drug carrier for targeted delivery of methotrexate (MTX) to tumors. Due to pathophysiological conditions in neoplastic tissue, high amounts of albumin accumulate in tumors and are metabolized by malignant cells. MTX, covalently coupled to human serum albumin (MTX-HSA) for cancer treatment, is currently being evaluated in phase II clinical trials. Because synovium of patients with rheumatoid arthritis (RA) shares various features observed also in tumors, albumin-based drug targeting of inflamed joints might be an attractive therapeutic approach. Therefore, the pharmacokinetics of albumin and MTX in a mouse model of arthritis was examined. Additionally, uptake of albumin by synovial fibroblasts of RA patients and the efficacy of MTX and MTX-HSA in arthritic mice were studied. The results show that when compared with MTX, significantly higher amounts of albumin accumulate in inflamed paws, and significantly lower amounts of albumin are found in the liver and the kidneys. The protein is metabolized by human synovial fibroblasts in vitro and in vivo. MTX-HSA was significantly more effective in suppression of the onset of arthritis in mice than was MTX. In conclusion, albumin appears to be a suitable drug carrier in RA, most likely due to effects on synovial fibroblasts, which might increase therapeutic efficacy and reduce side effects of MTX
    Type of Publication: Journal article published
    PubMed ID: 12707361
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  • 2
    Keywords: INVASION ; Germany ; human ; IN-VIVO ; MODEL ; VIVO ; MICE ; GENE-TRANSFER ; REDUCTION ; INDUCTION ; CONTRAST ; fibroblasts ; treatment ; MOUSE ; score ; IMMUNODEFICIENT MICE ; DEGRADATION ; MOUSE MODEL ; METHOTREXATE ; BEARING RATS ; albumin ; fibroblast ; rheumatoid arthritis ; RHEUMATOID-ARTHRITIS ; CYTOKINE INHIBITORS ; DESTRUCTION ; INTERLEUKIN-1
    Abstract: Objective: To investigate the effect of methotrexate (MTX) and albumin coupled with methotrexate (MTX-HSA) on cartilage invasion and induction of perichondrocytic degradation by rheumatoid arthritis synovial fibroblasts ( RA SF) in vivo. Methods: Human cartilage and RA SF were co-transplanted in three groups of severe combined immunodeficient mice ( SCID), which received 1 mg/kg free MTX (n = 9), 1 mg/kg MTX-HSA ( n = 6), or 0.9% NaCl ( n = 5), respectively, intraperitoneally twice a week. After 4 weeks' treatment, the mice were killed and the implants analysed histologically. Results: The control group had a mean (SEM) score for cartilage invasion of RA SF of 2.0 (0.26) and for perichondrocytic cartilage degradation of 1.5 (0.34). In contrast, mice which received MTX showed a significantly reduced invasion (0.78 (0.14), p〈 0.01) and a reduction in perichondrocytic cartilage degradation scores (0.69 (0.2), p〈 0.05) in comparison with the control group. Mice treated with MTX-HSA also had significantly reduced scores for RA SF invasion into the cartilage (0.92 (0.41), p〈 0.05) and for cartilage degradation (0.83 (0.44), p〈 0.05) compared with controls. The effects of MTX and MTX-HSA were not significantly different between these two groups. Conclusion: Treatment with MTX or MTX-HSA significantly ameliorates cartilage destruction in the SCID mouse model for human RA
    Type of Publication: Journal article published
    PubMed ID: 15194591
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  • 3
    ISSN: 1432-1440
    Keywords: Kollagen ; Immunfluorescenz ; Thrombus ; Blutplättchen ; Collagen ; immunofluorescence ; thrombus ; platelets
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Sections of arterial walls and of thrombi and smears of leukocytes previously incubated in vitro with collagen type III were examined by immunohistochemical technique for the presence of collagen types I, II and III. In arterial walls collagen type III was detected immediately underlaying the endothelial cell layer and in the tissue between tunica elastica interna and adventitia. Collagen type I was not shown in the subendothelial layer. Fresh thrombi contained occasionally collagen, but only of type III. This was associated with leukocytes. Leukocytes were capable in vitro to associate and/or phagocytose collagen type III and this could be visualized immunohistochemically. The data show that collagen type III in vivo may play a crucial role in the initiation of thrombus formation.
    Notes: Zusammenfassung Schnitte von arteriellen Gefäßwänden und von Thromben sowie Ausstriche von Leukocyten, die in vitro mit Kollagen Typ III inkubiert worden waren, wurden mit Hilfe der indirekten Immunfluorescenz auf die Verteilung und Anwesenheit von Kollagen Typ I, II und III untersucht. In Arterienwänden wurde Kollagen Typ III direkt subendothelial gefunden, sowie zwischen Tunica elastica interna und Adventitia. Kollagen Typ I war in der subendothelialen, der Tunica elastica interna direkt aufliegenden Schicht nicht nachweisbar. Frische Thromben enthielten gelegentlich Kollagen, jedoch nur Typ III. Dieses war mit Leukocyten assoziiert. Auch in vitro waren Leukocyten in der Lage, Kollagen Typ III zu assoziieren und/oder phagocytieren. Solches Kollagen war immunohistochemisch nachweisbar. Die Ergebnisse zeigen, daß Kollagen Typ III vermutlich eine entscheidende Rolle bei der arteriellen Thrombusbildung spielt.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-1440
    Keywords: Lebercirrhose ; Kollagen ; Immunfluorescenz ; Liver cirrhosis ; Collagen ; Immunofluorescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Pepsin solubilization of small and large noduled liver cirrhosis yielded two types of collagen (precipitated at 1.7 and 2.5 M NaCl concentrations) as demonstrated by electronmicroscopy. The 1.7 M NaCl precipitate was identified as type III collagen using an immunofluorescence technique. The 2.5 M NaCl precipitate appeared to be type I in the electronmicroscope. However, immunofluorescent and biochemical studies indicated that it was not type I but a type of collagen not yet described.
    Notes: Zusammenfassung Aus klein- und grobknotiger Lebercirrhose durch Pepsinbehandlung gelöstes Kollagen enthält zwei Kollagentypen (1,7 M und 2,5 M NaCl Präcipitat). Immunfluorescenzmikroskopisch konnte Kollagen Typ III, das dem 1,7 M NaCl Präcipitat entspricht, nachgewiesen werden. Das 2,5 M NaCl Präcipitat erscheint elektronenmikroskopisch als Typ I. Dagegen zeigen immunfluorescenzmikroskopische und biochemische Befunde, daß es sich hierbei nicht um Kollagen Typ I handelt, sondern um einen bisher noch nicht beschriebenen Kollagentyp.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-1440
    Keywords: Knorpel-Knochenumwandlung ; Osteoarthrose ; Chondrocyten ; Kollagensynthese ; Immunfluorescenz ; Cartilage-bone metamorphosis ; Osteoarthrosis ; Chondrocytes ; Collagen-synthesis ; Immunofluorescence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Synthesis of collagen by chondrocytes was studied by immunofluorescence using antibodies specific for type I, II and III collagen. The following tissues and culture conditions were chosen for this immunohistological study: normal articular cartilage, epiphyseal growth cartilage, cartilage undergoing osteoarthrotic degeneration, suspension culture and monolayer culture. While type II collagen is the unique collagen all over hyaline cartilage, type I collagen is produced by hypertrophic chondrocytes in the growth plate. In addition, chondrocytes in osteoarthrotic areas of articular cartilage synthesize type I collagen. Under in vitro culture conditions, chondrocytes initially produce type II collagen and synthesize later on type I collagen. The change of synthesis from type II to type I collagen is more rapid in monolayer than in suspension culture. It is concluded that the presence of matrix compounds and the cellmatrix interaction as well are necessary to maintain synthesis of type II collagen in chondrocytes. Alterations in the cell-matrix interactions are shown to occur in the hypertrophic zone of the epiphyseal growth plate, in cartilage undergoing osteoarthrotic degeneration as well as in chondrocytes grown in culture. Thus, change in the control of gene activity may subsequently lead to change in collagen synthesis. It is possible that the synthesis of type I collagen, which cannot fulfil the physiological function of a structural element in cartilageneous tissue, is a crucial factor in the process of osteoarthrosis.
    Notes: Zusammenfassung Unter Verwendung von spezifischen Antikörpern ist es möglich, mit Hilfe immunhistologischer Methoden die Verteilung der verschiedenen Kollagentypen und auch die Kollagensynthese einzelner Zellen zu verfolgen. Diese Methode wurde angewendet, um die Kollagensynthese von Chondrocyten im normalen Gelenkknorpel, Epiphysenknorpel der Wachstumsplatte, osteoarthrotischen Gelenkknorpel, sowie unter in vitro Kulturbedingungen zu verfolgen. Während im normalen Knorpel nur Typ II Kollagen synthetisiert wird, wurde festgestellt, daß die hypertrophierenden großen Chondrocyten an der Basis des Säulenknorpels auf die Synthese des Typ I Kollagens umschalten. Ähnliches gilt auch für die arthrotischen Zellen des Gelenkknorpels. Hier führt die Degeneration zu einer Umschaltung der Kollagensynthese von Typ II auf Typ I Kollagen. Dieses Phänomen kann auch in vitro nachvollzogen werden. In Suspensionskulturen, in denen Chondrocyten in Aggregaten eine knorpelähnliche Matrix aufzubauen vermögen, vollzieht sich der Umschaltungsprozeß langsamer als in Monolayerkulturen. Diese Beobachtungen zeigen, daß Chondrocyten zur Aufrechterhaltung ihrer Typ II Kollagensynthese eine spezielle Zell-Matrix-Wechselwirkung benötigen. Wird durch eine Veränderung der Knorpelmatrix (z.B. bei dem Degenerationsprozeß der Osteoarthrose) diese Wechselbeziehung gestört, kommt es zu einer Neuorientierung der Kollagensynthese und — in deren Folge — zur Umschaltung auf Typ I Kollagen.
    Type of Medium: Electronic Resource
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