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）： 尽管该领域已经取得了重大进展，但机械强度和长期存储方法的关键问题限制了工程船只的进展。众所周知，传统的冷冻保存会导致在细胞和周围细胞外基质中破坏冰的形成。在第一阶段的研究中，我们证明了无冰的冷冻保存（称为玻璃化）可以对多乙醇酸的冷冻保存产生巨大的影响，这些酸性酸的工程血管含有平滑肌细胞，这些血管含有平滑肌细胞，否则会因冷冻而受到重大伤害。与冷冻血管相比，新鲜的，冷冻保存的（冷冻）和玻璃化工程血管显示，新鲜和玻璃体的凋亡或坏死细胞几乎没有凋亡或坏死细胞。代谢测定结果表明，玻璃组织的活力与新鲜对照相似。玻璃化样品的收缩性结果超过82.7％的新鲜对照，在明显的对比度中，冷冻容器样品的结果仅占新鲜对照的10.7％（p <0.001）。对冷冻和玻璃化工程动脉的冷冻物质研究显示，玻璃化的标本中的冰可忽略不计，并且在冷冻标本的细胞外基质中形成了广泛的冰。被动机械测试显示冷冻保存后的组织强度增强。玻璃化是组织工程血管结构的可行储存方法。在此II阶段提案中，将玻璃化作为组织工程血管的储存方法的测试被扩展为包括内皮化和大型动物植入物研究。此外，将在体外筛选几种新的具有更好冰控制的玻璃化方法，并且将在第二年的大型动物模型中测试两种最有效的方法。最后，我们将优化一种用于人类组织血管的无冰冷冻保存技术。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用于细胞和组织存储的方法，使组织工程设备在美国和全球范围内都可以使用，无论环境条件如何。