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在生物反应器中通过脉动刺激构建弹性大肌性血管壁。

Engineering of an elastic large muscular vessel wall with pulsatile stimulation in bioreactor.

作者信息

Xu Zhi C, Zhang Wen J, Li Hong, Cui Lei, Cen Lian, Zhou Guang D, Liu Wei, Cao Yilin

机构信息

Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Tissue Engineering, Shanghai, China.

出版信息

Biomaterials. 2008 Apr;29(10):1464-72. doi: 10.1016/j.biomaterials.2007.11.037. Epub 2007 Dec 21.

Abstract

Tissue engineering offers a new approach for the construction of vascular substitutes in vitro with proper mechanical properties. Although success has been made in the engineering of small blood vessels (<6mm in diameter), it remains a challenge to engineer large vessels (>6mm in diameter) due to their insufficient biomechanical property. In the current study, an elastic large vessel wall (6mm in diameter) was engineered by loading a polyglycolic acid (PGA) unwoven fiber scaffold seeded with smooth muscle cells (SMCs) on a vessel reactor designed with dynamic culture conditions. SMCs were isolated from canine carotid artery and expanded before seeding on a PGA fiber mesh. The cell-seeded PGA mesh was then loaded on a vessel reactor and subjected to pulsatile stimuli. Grossly, an elastic vessel wall was formed after 8 weeks of dynamic engineering. Histological examination showed well-orientated smooth muscle cells and collagenous fibers in the group with dynamic culture. In addition, the phenotype of SMCs was confirmed by positive staining of smooth muscle alpha-actin and calponin. On the contrary, disorganized smooth muscle cells and collagenous fibers were observed in the group under static culture without stimuli. Furthermore, the engineered vessels under dynamic culture exhibited significant improvements on biomechanical property over the one from static culture. Our results indicate that the approach developed in the current work is efficient for large vessel engineering. This approach may also be suitable for the engineering of other tissues with muscular tubular structure.

摘要

组织工程学为体外构建具有适当机械性能的血管替代物提供了一种新方法。尽管在小血管(直径<6mm)工程方面已取得成功,但由于大血管(直径>6mm)生物力学性能不足,对其进行工程构建仍然是一项挑战。在当前研究中,通过将接种有平滑肌细胞(SMC)的聚乙醇酸(PGA)无纺纤维支架加载到设计有动态培养条件的血管反应器上,构建了一个弹性大血管壁(直径6mm)。从犬颈动脉分离出SMC并进行扩增,然后接种到PGA纤维网上。然后将接种细胞的PGA网加载到血管反应器上并施加脉动刺激。大体观察,动态工程8周后形成了弹性血管壁。组织学检查显示,动态培养组中平滑肌细胞和胶原纤维排列良好。此外,通过平滑肌α-肌动蛋白和钙调蛋白的阳性染色证实了SMC的表型。相反,在无刺激的静态培养组中观察到平滑肌细胞和胶原纤维排列紊乱。此外,动态培养的工程血管在生物力学性能方面比静态培养的血管有显著改善。我们的结果表明,当前工作中开发的方法对大血管工程是有效的。这种方法也可能适用于其他具有肌肉管状结构的组织工程。

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