Breakthrough Tissue and Organ Preservation and Transplantation Using Scaled-Up Nanowarming Technology

利用大规模纳米变暖技术实现突破性组织和器官保存和移植

• 批准号: 9757813
• 负责人: JOHN C BISCHOF
• 金额: $ 62.78万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2021-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9757813
• 关键词: Animal Model; Animals; Aorta; Architecture; Arteries; Biocompatible Materials; Biological; Blood Vessels; Cells; Cellular Structures; Convection; Coupling; Cryopreservation; Cryopreserved Tissue; Cryoprotective Agents; Crystal Formation; Crystallization; Data; Degenerative Disorder; Dehydration; Development; Devices; Electromagnetics; Engineering; Eye; Fracture; Freezing; Frequencies; Funding; Future; Glass; Health Care Costs; Heart; Heart Transplantation; Heating; Hour; Human; Ice; Kidney; Life; Link; Liquid substance; Lung; Magnetic nanoparticles; Magnetism; Measures; Methods; Modeling; Mus; National Heart, Lung, and Blood Institute; Neonatal; Nitrogen; Organ; Organ Donations; Organ Donor; Organ Preservation; Organ Transplantation; Oryctolagus cuniculus; Outcome; Patients; Performance; Polyethylene Glycols; Process; Production; Protocols documentation; Quality Control; Rattus; Recovery; Regenerative Medicine; Rewarming; Rodent; Sample Size; Sampling; Savings; Services; Silicon Dioxide; Speed; Stabilizing Agents; Structure; System; Techniques; Technology; Temperature; Testing; Time; Tissue Engineering; Tissue Preservation; Tissue Transplantation; Tissue Viability; Tissues; Toxic effect; Transplantation; Transplanted tissue; Transportation; attenuation; biomaterial compatibility; clinical translation; cold temperature; cryogenics; functional restoration; improved; in vivo; interest; iron oxide nanoparticle; magnetic field; nanomaterials; nanoparticle; nanoparticle delivery; nanowarming; new technology; novel; particle; prevent; radio frequency; response; scale up; success; thermal stress; tissue/organ preservation; transplant model; transplantation medicine; vitreous state

ABSTRACT: Rewarming biomaterials from the vitrified state is a critical step in obtaining successful cryopreservation. Successful techniques for rescuing cryopreserved bulk biomaterials and organs would not only provide critical improvements for donor-organ transport, supply, and matching, but is also a missing link in the potential supply chain for engineered tissues. Typical freezing processes cause significant damage to biomaterials through ice crystal formation and cellular dehydration. However, with the aid of cryoprotectant (CPA) solutions, biospecimens can be stabilized in the vitreous (i.e. “glass” or “amorphous”) state, allowing for long-term cryopreservation. A number of groups have employed successful techniques for cooling bulk systems to the vitreous state (including entire rabbit kidneys). Rewarming these vitrified biomaterials is a greater engineering challenge, due to the critical warming rates (hundreds of oC/min) necessary to avoid devitirification (i.e. crystallization) during thaw. In addition, non-uniformity in temperature field produces thermal stresses that can crack the brittle material, and so both speed and uniformity of thaw are of critical importance. Here we propose to investigate the ability of radiofrequency heated magnetic nanoparticles, or “nanowarming,” to overcome this major limitation hindering further development of bulk cryopreservation approaches. Although electromagnetic rewarming has been tried, the direct coupling of the waves to tissue inherently results in non-uniformity in heating, which leads to cracking and differential viability. At lower radiofrequencies (RF < 1 MHz) alternating magnetic fields (AMFs) can uniformly penetrate tissues without attenuation and negligible dielectric coupling. Although these lower frequency fields will be unable to rapidly heat the tissue on their own, they are ability to produce significant heating through coupling with magnetic (e.g. iron-oxide) nanoparticles. We have already demonstrated that this approach is able to generate heating rates rapid enough to avoid devitirification (greater than 200 oC/min) and should scale independent of sample size. The objective of this study is to refine this novel nanowarming technology for use in cryopreserving biologic tissues and intact organs for transplant. To this end, in Aim 1 we will scale up the nanoparticle production process and the size of the RF heating device. In Aim 2 we will optimize CPA and nanoparticle composition and loading/unloading conditions for vitrification and nanowarming of cells and tissues (arteries). In Aim 3 we will test these optimized conditions in heart transplant models of increasing size and complexity. In summary, the focus of this proposal will be to leverage our breakthrough nanowarming technology by optimizing CPA composition and nanoparticle delivery in a scaled up system capable of vitrifying and recovering cells, arteries, and intact organs with an eye on future application for cryopreserving tissues and organs for use in human transplantation.

抽象的： 将生物材料从玻璃化状态复温是获得成功冷冻保存的关键步骤。 拯救冷冻保存的散装生物材料和器官的成功技术不仅可以提供关键的 供体器官运输、供应和匹配方面的改进，但也是潜在供应中缺失的一环 工程组织链典型的冷冻过程会通过冰对生物材料造成严重损害。 晶体形成和细胞脱水然而，在冷冻保护剂（CPA）溶液的帮助下， 生物样本可以稳定在玻璃体（即“玻璃”或“无定形”）状态，从而可以长期保存 许多团体已经采用了成功的技术来冷却散装系统。 玻璃体状态（包括整个兔子肾脏）的复温是一项更大的工程。 挑战，由于避免失透所需的临界升温速率（数百摄氏度/分钟）（即。 融化过程中的结晶）此外，温度场的不均匀性会产生热应力。 使脆性材料破裂，因此解冻的速度和均匀性至关重要。 在这里，我们建议研究射频加热磁性纳米颗粒的能力，或者 “纳米变暖”，以克服阻碍批量冷冻进一步发展的主要限制 尽管已经尝试过电磁复温，但波与组织的直接耦合。 本质上会导致加热不均匀，从而导致开裂和活力较低。 射频 (RF < 1 MHz) 交变磁场 (AMF) 可以均匀地穿透组织，而无需 尽管这些较低频率的场将无法快速衰减和可忽略不计的介电耦合。 自行加热组织，它们能够通过与磁性（例如磁力）耦合产生显着的热量。 我们已经证明这种方法能够产生加热速率。 足够快以避免失透（大于 200 oC/min），并且应该独立于样品大小。 本研究的目的是完善这种新颖的纳米加热技术，用于冷冻保存生物制品 为此，在目标 1 中，我们将扩大纳米颗粒的生产规模。 在目标 2 中，我们将优化 CPA 和纳米颗粒的成分。 以及细胞和组织（动脉）玻璃化和纳米变暖的加载/卸载条件。 将在尺寸和复杂性不断增加的心脏移植模型中测试这些优化条件。 总之，该提案的重点是通过以下方式利用我们突破性的纳米变暖技术： 在能够玻璃化和固化的放大系统中优化 CPA 成分和纳米颗粒输送 恢复细胞、动脉和完整器官，着眼于冷冻组织的未来应用 用于人体移植的器官。