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The new attempt of reconstructed artificial urethra using VAMG

VAMG重建人工尿道的新尝试

基本信息

批准号：
15591695
负责人：
TOKUDA Yuji
金额：
$ 2.3万
依托单位：
Saga University (2004-2005)佐賀医科大学 (2003)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591695/
关键词：
orthotropic artificial urethra omentum urotheilum VAMG 尿略上皮中皮塩酸処理

项目摘要

Although the inflammatory reaction plays an important role in the tissue healing process, it causes adverse impact such as post injury scar. In this study, we examined the preventive effects of the reconstructed urothelium on the scaffold vascular acellular matrix graft (VAMG) against scar formation in rat. The bladder urothelium, Aorta and inferior vena cava were obtained from 10 weeks old male Wister rat. Chemically removed and mechanically dissociated urothelial cells were cultured for two weeks in culture dish. The VAMGs as the scaffold were prepared from Aorta and inferior vena cava wall by denucleation using 2000k units DNA ase and 4%sodium desoxycholeate. Cultured epithelial cells were implanted on the scaffold in culture system in vitro. After 7 days, these culture assemblies were put into the rat omentum pouch, where mesothelium had been chemically denuded, as in vivo environment with rich blood supply. As a control study, cultured epithelial cells were also implanted into the omentum pouch directly without VAMG. The process of epithelization on the VAMG was histologically examined 7 and 14 days after implantation. On day 7, reconstructed urothelia on the scaffold were partly alive with the reactive lymphocyte infiltration surrounding the VAMG. However, epithelial cells were not observed around the VAMG and the inside of VAMG was filled with granulation on day14. On the other hand, urothelial cell was not found but severe inflammatory reaction was observed in the omentum pouch on day 7 in control rats.Our present preliminary experiment demonstrated that the reconstructed urothelium on the VAMG might be able to prevent inflammatory reaction resulting scarred tissue in the injured urinary tract until the inherent urothelium is recovered.

虽然炎症反应在组织愈合过程中起着重要作用，但也会产生损伤后瘢痕等不良影响。在本研究中，我们研究了重建尿路上皮对支架血管脱细胞基质移植（VAMG）对大鼠瘢痕形成的预防作用。取10周龄雄性Wister大鼠膀胱尿路上皮、主动脉和下腔静脉。化学去除和机械分离的尿路上皮细胞在培养皿中培养两周。以主动脉和下腔静脉壁为材料，用200k单位DNA酶和4%去氧胆酸钠去核制备vamg作为支架。体外培养系统将培养的上皮细胞移植到支架上。7 d后，将这些培养体放入化学剥去间皮的大鼠网膜袋中，如体内血供丰富的环境。作为对照研究，培养的上皮细胞也不经VAMG直接植入大网膜袋。在移植后第7天和第14天分别对VAMG的上皮形成过程进行组织学检查。第7天，支架上重建的尿路上皮部分存活，反应性淋巴细胞浸润在VAMG周围。然而，在第14天，VAMG周围未见上皮细胞，VAMG内部充满肉芽。另一方面，对照组大鼠在第7天未发现尿路上皮细胞，但在大网膜袋内观察到严重的炎症反应。我们目前的初步实验表明，在原有的尿路上皮恢复之前，在VAMG上重建的尿路上皮可能能够防止炎症反应导致损伤的尿路组织瘢痕。