Transplantation of Schwann cells (SCs) is a promising treatment modality to improve neuronal regeneration. Identification of the transplanted cells is an important step when studying the development of this method. Genetic labeling is the most stable and reliable method of cell identification, but it is still unclear whether it has deleterious effect on SC characteristics. Our aim was to achieve a stable population of SCs transduced with the lacZ gene at a high frequency using a retroviral vector in vitro, and to follow the labeled SC in vitro to assess their viability and phenotypic marker expression. Furthermore, we transplanted lacZ-labeled SCs in a conduit to repair peripheral nerve to investigate their effect on nerve regeneration in vivo. Rat and human SCs were cultured and transduced with an MFG lacZ nls marker gene, achieving a transduction rate of 80% and 70%, respectively. Rat SCs were kept in culture for 27 weeks and examined every 4 weeks for expression of lacZ, viability, and phenotypic marker expression of GFAP, p75, MHC I and II. Throughout this period, transduced rat SCs remained viable and continued to proliferate. The proportion of cells expressing lacZ dropped only by 10% and the expression of phenotypic markers remained stable. Transduced human SCs were followed up for 4 weeks in culture. They proliferated and continued to express the lacZ gene and phenotypic marker expression of GFAP and p75 was preserved. Primary culture of transduced rat SCs were transplanted, syngeneically, in a conduit to bridge a 10 mm gap in sciatic nerve and the grafts were examined after 3 weeks for the presence and participation of labeled SCs and for axonal regeneration distance. Transplanted transduced rat SCs were clearly identified, taking part in the regeneration process and enhancing the axonal regeneration rate by 100% (at the optimal concentration) compared to conduits without SCs. Thus, retroviral introduction of lacZ gene has no deleterious effect on SCs in vitro and these SCs take part and enhance nerve regeneration in vivo. GLIA 34:8-17, 2001.
The authors report a case of septic pericardial effusion resulting in cardiac tamponade associated with intrathoracic botryomycosis in a dog. Septic pericarditis and a pulmonary mass were diagnosed, and subtotal pericardiectomy and lobectomy of the affected pulmonary areas were carried out. Histopathology of the excised tissue showed changes supportive of botryomycosis--namely a pyogranulomatous inflammation with neutrophils centred around amorphous homogeneous eosinophilic material and club-like bodies containing Gram-positive bacterial cocci present in the centre. The patient recovered well following surgery and antibiotic therapy. To the authors' knowledge, this is the first report of pulmonary botryomycosis in the dog and the first report of this condition presented with pericardial involvement and cardiac tamponade in any species.
Cultured epithelial autografts are often applied to wounds with a capacity for regeneration from dermal appendages. It is unclear in these circumstances whether the cultured autografts act merely as a biologic dressing or whether they become incorporated into the new epithelium. We have used retroviral gene transfer techniques to identify autologous keratinocytes in an established porcine model of cultured epidermal (CE) grafting. Porcine keratinocytes were transduced with an MFG-lacZ nls vector produced by the amphotropic packaging line GP+EnvAm12. Transduction rates of 15.1%, in the absence of selection, were achieved by a single passage on gamma-irradiated retroviral producers as a feeder layer. Full-thickness wounds were created on Large White pigs and isolated from the surrounding skin by a polytetrafluoroethylene chamber. Wounds were grafted initially with autologous de-epidermized dermis (DED), followed 7 d later by sheets of retrovirally marked or unmarked CE autografts. Two weeks after grafting, the mean area of epithelium was 48.4% in wounds that received CE grafts and 32.3% in wounds that were left as DED alone. The epithelium on DED represents regeneration from dermal appendages. The contribution made by the autograft cells to the new epidermis was demonstrated unequivocally, however, by lacZ-positive areas visible macroscopically on the surface of the excised wound. In cryostat sections through the lacZ-positive areas, retrovirally marked cells were present at both superficial and basal positions in the new epithelium.
A new technique for the treatment of severe epistaxis associated with hereditary haemorrhagic telangiectasia is described. The nasal septum and inferior turbinates, surgically denuded of respiratory epithelium, were grafted using autografts of cultured epithelial sheets derived from buccal epithelium. All patients upon whom this technique has been used have shown considerable lessening in the frequency and severity of their epistaxes although two patients received grafts on two occasions, in each case approximately three months apart. It is postulated that a nasal lining of stratified squamous epithelium is likely to be more resistant to trauma than the normal respiratory type, and this is supported by the observation that bleeds very seldom occur from the oral cavity in this syndrome.
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