Dermal substitutes can be used to improve the wound healing of deep burns when placed underneath expanded, thin autologous skin grafts. Such dermal matrix material can be derived from xenogeneic or human tissue. Antigenic structures, such as cells and hairs must be removed to avoid adverse inflammatory response after implantation. In this study, a cost-effective method using low concentrations of NaOH for the de-cellularization of human donor skin preserved in 85% glycerol is described. The donor skin was incubated into NaOH for different time periods; 2, 4, 6 or 8 weeks. These dermal matrix prototypes were analyzed using standard histology techniques. Functional tests were performed in a rat subcutaneous implant model and in a porcine transplantation model; the prototypes were placed in full thickness excision wounds covered with autologous skin grafts.An incubation period of 6 weeks was most optimal, longer periods caused damage to the collagen fibers. Elastin fibers were well preserved. All prototypes showed intact biocompatibility in the rat model by the presence of ingrowing blood vessels and fibroblasts at 4 weeks after implantation. An inflammatory response was observed in the prototypes that were treated for only 2 or 4 weeks with NaOH. The prototypes treated with 6 or 8 weeks NaOH were capable to reduce wound contraction in the porcine model. In neo-dermis of these wounds, elastin fibers derived from the prototype could be observed at 8 weeks after operation, surrounded by more random orientated collagen fibers. Thus, using this effective low cost method, a dermal matrix can be obtained from human donor skin. Further clinical studies will be performed to test this material for dermal substitution in deep (burn) wounds.
Background Milky spots have been described as reactive structures, their classification varying from inflamed or haematopoietic tissue to lymphoid organs. In this study we investigated the reactivity of the milky spots in the omentum of rats upon induction of a chronic immune response in the peritoneal cavity. Methods At different time points after intraperitoneal administration of Bacillus Calmette‐Guérin (BCG), a peritoneal lavage was made, and the omentum and the draining parathymic lymph nodes were taken out. The cellular composition of these tissues was examined on the light microscopic level, using a panel of monoclonal antibodies, and also by electron microscopy. Results During the first 4 months after administering BCG, the number and size of the milky spots increased enormously. Separate macrophage, T, and B cell areas were formed, but interdigitating cells and follicular dendritic cells were not observed. The number of cells in the peritoneal cavity also increased, and the cellular composition showed a strong similarity with that of the milky spots. Especially during the onset of the experiment, most bacteria were observed in the macrophages in the milky spots rather than in the draining lymph nodes. A cellular immune response was observed in the parathymic lymph nodes but not in the milky spots. Conclusions Milky spots, either unstimulated or stimulated, should be classified as perivascular infiltrates. They play a role in the initial clearance of bacteria from the peritoneal cavity. Although the large increase in cell number is predominantly caused by immigration of cells, the results do support the role of milky spots as a site for local proliferation and maturation of especially macrophages and also B cells. The obtained data, however, do not support the earlier made assumption that milky spots function as a secondary lymphoid organ in the peritoneal cavity. © 1996 Wiley‐Liss, Inc.
The morphology and kinetics of macrophages and reticulum cells of rat lymph nodes have been studied in relation to the immune response to a second exposure to antigen. During the first 24 h after stimulation monocyte-like exudate macrophages, including some scattered interdigitating cells (IDC), contain granules similar to those present in epidermal Langerhans cells and lymph-borne veiled cells. In this induction phase these macrophages migrate from the marginal sinus into the paracortex and during the migration they gradually transform into IDC. In the proliferation phase the paracortex is mainly populated by transitional macrophages and there are almost no typical IDC present between the lymphoblasts. In the memory phase the relative number of IDC again rapidly increases. During this period in the paracortex there are often typical IDC which contain partially digested necrotic lymphocytes, thus resembling tingible body macrophages (TBM) of the germinal centre in this respect. It is suggested that the newly arrived macrophages induce the lymphoblast reaction, while mature IDC may have an inhibitory function in the memory phase of the immune response. In this phase the phagocytic potential of IDC is clearly shown.
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