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Research of muscle tissue engineering for urethral sphincter

尿道括约肌肌肉组织工程研究

基本信息

批准号：
16591595
负责人：
YAMADA Yuji
金额：
$ 1.9万
依托单位：
Kobe University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16591595/
关键词：
Tissue Engineering angiogenesis urology 再生成長因子

项目摘要

To achieve the goals of engineering large complex tissues, and possibly internal organs, vascularization of the regenerating tissue is essential. To keep an initial volume after implantation of regenerated tissue, improved vascularization is considered to be important. Recent advances in understanding the process of blood vessel growth has offered significant tools for the neovascularization of bioengineered tissues and therapeutic angiogenesis. Several angiogenic growth factors including vascular endothelial cell growth factor (VEGF), basic fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF), platelet derived growth factor (PDGF) were used for vascularization of ischemic tissues. Other approaches such as prevascularization of the matrix prior to cell seeding and incorporation of endothelial cells in the bioengineered tissues showed encouraging results. In this study, the engineered tissues were shown to form muscle, as evidenced by H&E staining and immunohistochemical probing. Neovascularization was detected in the engineered muscle tissues of VEGF-infected cells. EC were shown to participate in blood vessel formation by X-gal staining. In contrast, engineered muscle of non-infected cells had a significantly smaller mass of cells, less neovascularization and only a few HMEC-1 cells could be detected. The VEGF-expressing tissues maintained their initial volume for 8 weeks, whereas the non-infected tissues lost their volume significantly. Sustained expression of muscle specific genes, myosin and MyoD, was evident only in muscle tissues containing VEGF-infected myoblasts. These results show, for the first time, that a combination of VEGF and endothelial cells are useful for inducing muscle formation, neovascularization and volume preservation in engineered tissues, and this technique may be useful for urinary sphincter reconstruction.

为了实现大型复杂组织和可能的内部器官的工程目标，再生组织的血管化是必不可少的。为了保持再生组织植入后的初始体积，改善血管化被认为是重要的。血管生长过程的最新进展为生物工程组织的新生血管和治疗性血管生成提供了重要的工具。血管生成生长因子包括血管内皮细胞生长因子（VEGF）、碱性成纤维细胞生长因子（bFGF）和肝细胞生长因子（HGF）、血小板衍生生长因子（PDGF）用于缺血组织的血管生成。其他方法，如细胞播种前基质预血管化和生物工程组织中内皮细胞的掺入，显示出令人鼓舞的结果。在本研究中，H&E染色和免疫组织化学探针证实，工程组织形成肌肉。在vegf感染细胞的工程肌肉组织中检测到新生血管。X-gal染色显示EC参与血管形成。相比之下，未感染细胞的工程肌肉细胞质量明显较小，新生血管较少，仅能检测到少量HMEC-1细胞。vegf表达组织的初始体积维持了8周，而未感染组织的体积明显下降。肌肉特异性基因肌球蛋白和MyoD的持续表达仅在含有vegf感染的成肌细胞的肌肉组织中明显。这些结果首次表明，VEGF和内皮细胞的结合在诱导肌肉形成、新生血管形成和工程组织的体积保存方面是有用的，并且这项技术可能对尿道括约肌重建有用。