| BackgroundCongenital hypospadias and urethral injury, urethral stricture is common disease of uropoiesis surgical department, its effective solution is still dependent on the surgical treatment, and the treatment of urethra shaping and revascularization is the main treatment modality.Urethroplasty and revascularization in more categories,there are currently nearly 300 species, and many academics and medical workers have been on the improvement and innovation. However, for long-urethral defect of complex urethral stricture and modalities of patients with urethral injury, postoperative complications such as urinary fistula, urethral stricture also remains high, so urologists become quite a headache.Research shows that the blood supply to the urethra and the occurrence of postoperative complications described above are also present in some connection.With the development of tissue engineering and stem cell technology, the ideal scaffolds for urethral reconstruction and good seed cells in basic and clinical research has achieved satisfactory results.Adipose stem cells sources of more extensive and easily obtained, and has a strong ability to regenerate bone tissue, cartilage tissue, myocardial tissue and epithelial tissue differentiation potential, etc, so that in the field of tissue engineering seed cells Research has gradually become a research focus of many researchers.Meanwhile, the vascular endothelial cells is a multifunctional cell, which located in the blood vessel wall and participated the pathogenesis of many of the body.A large number of animal experiments and clinical data indicate that, VEC has a unique advantage in improving the ischemic area and the body's blood supply to the injured site. In recent years, The reported of ADSCs and VEC separately United scaffold used to repair the urethra have increased gradually, and achieved certain results. However, there has not yet been reported on urethral reconstruction with the application of adipose derived stem cells and vascular endothelial cells co-culture in vain to repair rabbit urethral defect. ObjectsThe proposed of the experiments by applying the adipose derived stem cells and vascular endothelial cells co-culture in vitro, and acellular matrix material to build artificial urinary urethral defect repair. For endothelial cells to promote adipose stem cells in the blood vessels in urethra reconstruction has carried on the preliminary exploration. Methods1.Isolation, culture, amplification of rabbit adipose stem cells, and were identified by flow cytometry;Vascular endothelial cells are extracted, the culture and identification;ADSCs were co-cultured with the VEC.2.Respectively ADSCs, VEC and the two co-cultured cells were seeded to acellular matrix, tissue engineering urethra.3.The production of 60 male adult New Zealand rabbit randomly divided into five groups which urethral defect length 2.0cm animal model, the application.A group(cell-free control group) that is: a simple application of acellular matrix repair group.Group B(sham group) namely: urethral defect model for creating successful, and the two ends of the urethra urethral defect after direct anastomosis; group C(ADSCs group) namely: ADSCs acellular matrix composite surgical repair group; group D(VEC group) namely: VEC acellular matrix surgical repair group; E group(ADSCs and VEC co-culture group) namely: ADSCs VEC co-cultured with acellular matrix composite surgical repair group.4.On the first postoperative 1W,2W, 4W, 12 W were taken A, urethral reconstruction organization B, C, D, E group of experimental animals, histological and immunohistochemical observation and analysis.5. Postoperative 12 W, respectively, to retrograde urethrectomy angiography revealed several groups of experimental animals. Results1.In vitro amplification of ADSCs fusiform. Flow cytometry phenotype CD44, CD105 positive and CD34 negative, in line with mesenchymal stem cell characteristics; VEC primary adherent cells were grown in cell morphology diversity, round, oval, spindle-shaped or polygonal, passaged endothelial cobblestone-like; cells co-cultured for 7 days and some cells still retain the original spindle cell morphology may be in another part of the polygon, nest-like distribution, between 14 days interconnected cells can be seen in many synapses, some cells may fused into a lump.2.E group detecting urothelial vascular endothelial growth factor(F?), smooth muscle actin(a-SMA) were positive; respectively acellular matrix composite are growing well.3.Postoperative 1W,2W, 4W, 12 W, respectively, for each rabbit urethra group histological observation, the number of capillaries visible E group was significantly more than the C, D group;4.Statistical results show that the C, D, E, three groups compared with group A reduced risk of postoperative complications;And C, D, E incidence of postoperative complications are compared between three groups have no obvious difference. Conclusions1. ADSCs and VEC in vitro co-culture as seed cells can be constructed urethra of long urethral defect in tissue engineering;2. VEC supports the direction of change urothelial ADSCs, and can also accelerate the occurrence of blood vessels;... |
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