VITRIFICATION FOR TISSUE ENGINEERED BLOOD VESSELS

组织工程血管的玻璃化

• 批准号: 6735495
• 负责人: Ying Song
• 金额: $ 39.05万
• 依托单位: ORGAN RECOVERY SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6735495
• 关键词: biomechanics; blood vessel transplantation; blood vessels; carotid artery; cryopreservation; extracellular matrix; human tissue; swine; technology /technique development; tissue /organ preservation; tissue engineering

DESCRIPTION (provided by applicant): Although significant advances have been made in the field, the critical issues of mechanical strength and long-term storage methods limit progress in engineered vessels. It is well established that traditional cryopreservation results in damaging ice formation, both in the cells and in the surrounding extracellular matrix. In Phase I studies, we demonstrated that ice-free cryopreservation, known as vitrification, can have a salutary effects on the cryopreservation of polyglycolic acid - derived engineered vessels containing smooth muscle cells which otherwise sustain significant injury from freezing. Fresh, traditionally cryopreserved (frozen), and vitrified engineered vessels evaluated for apoptosis revealed few apoptotic or necrotic cells in fresh and vitrified vessels compared with frozen vessels. The metabolic assay results indicated that vitrified tissue had similar viability to fresh controls. The contractility results for vitrified samples were over 82.7% of fresh controls and in marked contrast, the results for frozen vessel samples were only 10.7% of fresh controls (p<0.001). Cryosubstitution studies of frozen and vitrified engineered arteries revealed negligible ice in the vitrified specimens, and extensive ice formation in the extracellular matrix of frozen specimens. Passive mechanical testing revealed enhanced tissue strength after cryopreservation. Vitrification is a feasible storage method for tissue engineered blood vessel constructs. In this Phase II proposal, testing of vitrification as a storage method for tissue engineered blood vessels is extended to include endothelialization and large animal implant studies. In addition, several new methods of vitrification with better ice control will be screened in vitro and the two most effective methods will be tested in a large animal model in year 2. Finally, we will optimize a technique for ice-free cryopreservation of human tissue engineered blood vessels. In Phase III, these studies will be extended to include clinical trials. The long-term goal of this proposed project is to develop methods for cell and tissue storage that will make it possible for tissue engineered devices to be available in the United States and worldwide, regardless of environmental conditions.

描述（由申请人提供）： 尽管该领域已取得重大进展，但机械强度和长期储存方法的关键问题限制了工程容器的进展。众所周知，传统的冷冻保存会导致细胞和周围细胞外基质中的冰形成受到破坏。在第一阶段的研究中，我们证明了无冰冷冻保存（称为玻璃化）可以对含有平滑肌细胞的聚乙醇酸衍生工程血管的冷冻保存产生有益的影响，否则这些血管会因冷冻而遭受严重损伤。对新鲜、传统低温保存（冷冻）和玻璃化工程血管进行细胞凋亡评估后发现，与冷冻血管相比，新鲜和玻璃化血管中几乎没有凋亡或坏死细胞。代谢测定结果表明玻璃化组织与新鲜对照具有相似的活力。玻璃化样品的收缩性结果超过新鲜对照的 82.7%，形成鲜明对比的是，冷冻容器样品的结果仅为新鲜对照的 10.7% (p<0.001)。对冷冻和玻璃化工程动脉的冷冻替代研究表明，玻璃化标本中的冰可以忽略不计，而冷冻标本的细胞外基质中则有大量的冰形成。被动机械测试显示冷冻保存后组织强度增强。玻璃化冷冻是组织工程血管构建体的一种可行的储存方法。在这个二期提案中，玻璃化作为组织工程血管储存方法的测试扩展到包括内皮化和大型动物植入研究。此外，还将在体外筛选几种冰控效果更好的玻璃化冷冻新方法，并在第二年在大型动物模型中测试两种最有效的方法。最后，我们将优化人体组织无冰冻存技术工程血管。在第三阶段，这些研究将扩展到包括临床试验。该拟议项目的长期目标是开发细胞和组织储存方法，使组织工程设备能够在美国和全世界范围内使用，无论环境条件如何。