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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK; Malden, USA : Blackwell Science Ltd/Inc.
    Wound repair and regeneration 12 (2004), S. 0 
    ISSN: 1524-475X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Human mesenchymal stem cells (hMSCs) obtained from a single donor from an iliac crest, were investigated with cutaneous wound healing models using nude rat, eliminating T cell-mediated immune reaction to the grafted stem cell of human origin. Co-culture of the human keratinocytes and the hMSCs revealed tight basal membranous interaction by electron microscopy.1.5 × 1.5 cm2 dorsal full-thickness defects including panniculus carnosus of F344/NJCl-rnu nude rats were covered by bi-layered porcine-derived collagen sponge artificial skin substitutes, Pelnac®, Johnson & Johnson, Tokyo, Japan, impregnated with 5 × 106 hMSCs grated to along with 0, 1, 10, or 100 μg of recombinant human basic fibroblast growth factor (bFGF). The tissues were harvested at 3, 7 42 days after grafting. The defects were remarkably epithelialized by 7 days after coverage with artificial skin substitutes and 10 μg of bFGF, while artificial substitutes with no hMSCs or artificial skin substitutes with hMSCs alone did not demonstrate such healing. The architectures of the artificial skin substitutes were integrated by day 42 in hMSCs-treated groups. The artificial skin substitute at least plays a role in as a template or scaffold of the grafted hMSCs. Up to day 3, the mesenchymal stem cell surface markers such as CD 29 and CD 44 were remained immunopositive in the groups with hMSCs-treated groups over the reconstructed dermis-like tissue. By 42 days after grafting, artificial skin substitutes with hMSCs and 10 μg of bFGF demonstrated the total epithelization and the keratinocytes by this treatment exhibited the pan-cytokeratin of human origin by immunohistochemical expressions. Thus, the hMSCs with bFGF treatment using an artificial skin substitute as a successfully heal. These results suggest that the human mesenchymal stem cells may be utilized for wound coverage together with bFGF and artificial skin substitutes.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford, UK; Malden, USA : Blackwell Science Inc
    Wound repair and regeneration 13 (2005), S. 0 
    ISSN: 1524-475X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Cell-to–cell interactions between human mesenchymal stem cells and potential adjacent cells such as endothelial cells, dermal fibroblasts, and epidermal keratinocytes was investigated. A modified dual Boyden chamber assay using 8-µm pores revealed a more powerful chemotactic cell migration of human mesenchymal stem cells toward human epidermal keratinocytes than other cells, such as umbilical artery endothelial cells and dermal fibroblasts, during 16 hours of incubation (336.2 ± 52.33, 36.0 ± 11.20, and 62.7 ± 18.16, cells/field, respectively, p 〈 0.01; comparison between endothelial cells and keratinocytes, and fibroblasts and keratinocytes). Scanning electron microscopy showed human mesenchymal stem cell migration through the pores, with endothelial cells, fibroblasts, or keratinocytes in the lower chambers. Mesenchymal stem cell ultrastructural changes occurred, including a larger euchromatin nucleus, when the cells were placed in medium containing 10 percent fetal bovine serum, whereas basic fibroblast growth factor maintained the immature cell morphology for 4 days. Monolayer coculture also showed human mesenchymal stem cell changes in ultrastructural morphology in the vicinity of the epidermal keratinocytes. These data suggest that human mesenchymal stem cells may interact with human epidermal keratinocytes to accelerate wound healing and coverage.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK; Malden, USA : Blackwell Science Inc
    Wound repair and regeneration 13 (2005), S. 0 
    ISSN: 1524-475X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: In vivo studies using bone marrow-derived mesenchymal stem cells are still uncommon. Applications for bone defect replacement in undesirable clinical circumstances such as large defects, bacterial or other pathogen-contaminated fields, and irradiated surgical wound bed necessitate vascularized bone regeneration. Use of a fascial flap including regenerated bone would be a very powerful tool for treatment. It would be especially beneficial in cases where normal bone regeneration is not expected due to a lack of sufficient blood supply, extensive surgical scaring, or bacterial contamination. In this study, we used nude rats in which the superficial epigastric flap of the experimental group was used to wrap around a mixture of human mesenchymal stem cells, bone morphogenetic protein-2, and basic fibroblast growth factor cytokines in a gelatin carrier. These rats showed significantly higher bone mineral density at 4 weeks compared to the other experimental groups containing phosphate buffered saline, human mesenchymal stem cells alone, or the two cytokines alone (p 〈 0.01). There were no remarkable histologic differences up to 7 days. At 2 weeks, more progressive vascularity and perivascular tissue deposits were seen in the experimental group. Basophilic mineral structure surrounded the fibroblast-like mesenchymal stem cells at 4 weeks, presumably osteoblastic or osteoclastic cell lining. Bone marker immunohistochemistry against alkaline phosphatase and osteocalcin revealed diffuse and distinct immunoreactivity in osteoblastic cells in the experimental group at 4 weeks. Further transcriptional expression of polyomavirus enhancer binding protein 2αA suggested that the human transplanted cells proceeded to osteogenic lineage in 4 weeks. These results may be useful as a new approach for bone regeneration.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Oxford, UK; Malden, USA : Blackwell Publishing Ltd/Inc.
    Wound repair and regeneration 13 (2005), S. 0 
    ISSN: 1524-475X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Acute wounds caused by the severe and extensive infection such as alpha-hemolytic streptococci or MRSA may be life-threatening in immunocompromised hosts such as steroid users or those with venostasis in the lower legs. The principle of the treatment is the removal of the primary pathogens, however, difficulties of the lower leg reconstruction remain due to the tissue loss and the deficiency of the circulation system after extensive soft tissue debridement. For 10 immediately-developing extensive lower extremity cases, artificial skin substitutes composed of dried bilayer membranes of outer silicone membrane and inner porcine-tendon derived collagen layer (Pelnac®, Gunze Co., Ltd., Kyoto, Japan) were immediately after extensive and meticulous debridement. While the skin substitute integrated into the wound bed for maximally 3 weeks, total 20 micrograms of the basic fibroblast growth factors (bFGF)(Trafermin®, Kaken Pharmaceutical Co. Ltd, Tokyo, Japan) were applied by the 27-Gage syringe daily. The outer membranes were removed and thin partial thickness skin grafting (9–10/1,000 inches) was performed and wound healed uneventfully.The bFGF-treated skin texture was significantly softer compared to control by a durometer (Teclock®, GS-701N, Tokyo, Japan)(p 〈 0.01), which was a measuring device elastic hard products such as rubbers and polymers.Reconstruction using the porcine-derived artificial skin substitute in the lower extremities after extensive debridement was safe, easy, and reproducible. The quality of the wounds with bFGF significantly improves the healed skin texture.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Malden, USA : Blackwell Science Inc
    Wound repair and regeneration 11 (2003), S. 0 
    ISSN: 1524-475X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Human mesenchymal stem cells obtained from the iliac crest of a single donor were investigated for cell proliferation, cell cycle profile, gene expression, and ultrastructural changes using electron microscopy. The human mesenchymal stem cells significantly increased their cell number by day 2 after treatment with bone morphogenetic protein-2 alone, or basic fibroblast growth factor alone or combinations of both proteins under serum-free conditions (p 〈 0.01). The human mesenchymal stem cells showed marked expression of cell nuclear antigen, notably at day 1, and pituitary tumor transforming gene throughout the experiment, suggesting cell cycle progression by bone morphogenetic protein-2 treatment. In addition, strong cellular nuclear bromodeoxyuridine incorporation was seen by immunocytochemistry. Fluorescence-activated cell sorting also showed a similar pattern of cell cycle progression with bone morphogenetic protein-2 treatment in serum-free medium and 10% fetal bovine serum treatment. The bone morphogenetic protein-2–treated human mesenchymal stem cells showed heterochromatin in the nucleus, suggesting cell differentiation, and well-developed granular endoplasmic reticulum, indicative of protein production. Overall, the human mesenchymal stem cells successfully proliferated with appropriate cell cycle progression and the cell ultrastructural morphology suggested marked nuclear and granular endoplasmic reticulum induction by bone morphogenetic protein-2 treatment in serum-free medium. (WOUND REP REG 2003;11:354–360)
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1524-475X
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: To facilitate bone healing in difficult circumstances, and to replace conventional therapeutic modalities, highly purified bone marrow-derived human mesenchymal stem cells (hMSCs) were investigated for induction of their osteogenic lineage upon provision of cytokine cues in vitro and in the cranial defect model in vivo. Alkaline phosphatase-expressing cells were most frequently observed when the hMSCs were treated with 2.5 ng/ml of basic fibroblast growth factor (bFGF) and 50 ng/ml of bone morphogenetic protein (BMP)-2 for 4 days in culture after a 6-day incubation in osteogenic medium containing dexamethasone, ascorbic acid-2-phosphate, and β-glycerophosphate. Four-millimeter full-thickness cranial defect wounds were made in male nude rats (F344/NJCl-rnu), whose deficit in the T cell compartment prevented T-cell–mediated cellular rejection. The animals were treated for 4 weeks with hMSCs and application of 10 µg each of bFGF and BMP-2 that had been soaked into a gelatin sponge carrier. Significant bone mineral density was observed by dual X-ray absorptiometry and this treatment also produced histologically mature osteocytes surrounded by both osteoblasts and osteoclasts expressing alkaline phosphatase and osteocalcin. The bone mineral densities and histological structures were matched at 8 weeks post-transplantation. Therefore, human bone marrow-derived mesenchymal stem cells are able to differentiate into an osteogenic lineage upon cytokine stimulation and accelerate healing in a nude rat cranial bone healing model.
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-6830
    Keywords: saccus vasculosus ; nucleus of the saccus vasculosus ; choroid plexus ; parathyroid hormone-related peptide (PTHrP) ; PTHrP receptor ; red stingray ; immunohistochemistry ; immunoradiometric assay
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 1. The exact role of the parathyroid hormone-related peptide (PTHrP) is not fully understood. We used immunohistochemistry to localize the PTHrP and its receptor in the brain of the red stingray, particularly in the saccus vasculosus (SV) and choroid plexus. 2. Immunoreactive PTHrP and its receptor were detected in the epithelial cells of the SV and the choroid plexus. In addition, the neuronal perikarya in the nucleus of the SV located in the hypothalamus is positive for the PTHrP. 3. No PTHrP-containing neurons were detected in the choroid plexus. Extracts of SV and choroid plexus showed positive reactions against the PTHrP and its receptor antibody in Western blot analysis. 4. High levels of immunoreactive PTHrP were detected in the plasma equivalent to those present in human humoral malignant hypercalcemia. In contrast, the immunoreactive PTHrP concentration in the cerebrospinal fluid was below detectable levels. 5. Our results suggest that the regulation of the PTHrP in the SV differs from that in the choroid plexus in the red stingray.
    Type of Medium: Electronic Resource
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