Ice-free vitrification and nanowarming of meniscal grafts for transplantation

用于移植的半月板移植物的无冰玻璃化和纳米加温

• 批准号: 10819333
• 负责人: Kelvin G.M. Brockbank
• 金额: $ 30.2万
• 依托单位: TISSUE TESTING TECHNOLOGIES, LLC
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-20 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10819333
• 关键词: AlamarBlue; Allogenic; Allografting; Animal Model; Animals; Biocompatible Materials; Biological Assay; Biomechanics; Cardiovascular system; Cartilage; Cell Culture Techniques; Cell Maintenance; Cell Survival; Cells; Chemistry; Chondrocytes; Convection; Coupling; Cryopreservation; Crystallization; Data; Diameter; Dimethyl Sulfoxide; Effectiveness; Evaluation; Extracellular Matrix; Family suidae; Fibrocartilages; Food Processing; Formulation; Freezing; Frequencies; Future; Gene Expression; Gene Expression Profiling; Harvest; Heart Valves; Heating; Human; IACUC; Ice; In Vitro; Investigation; Iron; Ischemia; Knee; Lateral meniscus structure; Life; Magnetic nanoparticles; Magnetism; Medial meniscus structure; Meniscus structure of joint; Metabolic; Methods; Operative Surgical Procedures; Outcome; Patient-Focused Outcomes; Patients; Penetration; Permeability; Pharmacopoeias; Phase; Plants; Property; Publications; Research; Review Literature; Rewarming; Sample Size; Sampling; Side; Small Business Innovation Research Grant; Stains; Surveys; Technology; Testing; Thick; Thinness; Tissue Engineering; Tissue Sample; Tissue Viability; Tissues; Transplantation; Trypan Blue; United States; articular cartilage; attenuation; biomaterial compatibility; clinical application; commercialization; experimental group; in vivo; innovation; nanoparticle; nanowarming; particle; preservation; radio frequency; reconstruction; repaired; translation to humans

ABSTRACT: Limited availability of fresh viable donor meniscal grafts reduces size-matching efficiency for meniscal allograft transplantation, restricting surgical accessibility and patient outcomes. Current preservation methods with retention of meniscal viability are inadequate. Fibrochondrocyte viability of transplanted menisci is accepted as one of the determinants of outcome following allograft transplantation. We have previously developed an ice-free vitrification method of cryopreservation that maintains excellent chondrocyte viability in animal models and human articular cartilage. We present preliminary data here demonstrating that fibrochondrocytes in menisci can also survive vitrification due to the absence of ice formation during cooling and warming of small 1-3mL samples. However, it has not been possible to rewarm larger samples due to ice nucleation during rewarming that results in loss of cell viability. The innovation in this proposal relates to a new rewarming method that does not have the limitations of boundary convection warming that should be effective for samples up to 50mL in volume. This rewarming method utilizes radio frequency induced heating of magnetic iron nanoparticles. We will optimize nanowarming of full thickness meniscal fibrocartilage plugs for maintenance of cell viability, including gene expression, and extracellular matrix integrity. This objective will be developed in two specific aims to optimize nanowarming variables and scaleup from 5mL employing tissue plugs to 30mL with intact menisci. Menisci will be vitrified and stored for 1-4 weeks and then nanowarmed meniscal cell viability, chemistry, and biomaterial properties will be compared with untreated fresh control tissue. The nanowarming conditions that provides the best preservation of cells with minimal if any biomaterial changes will be selected for further investigation in vivo and translation to human menisci in a subsequent, future, Phase II SBIR application.

摘要：新鲜活体半月板移植物的可获得性有限降低了手术的大小匹配效率 同种异体半月板移植，限制了手术的可及性和患者的预后。当前保存 保留半月板存活的方法是不够的。半月板移植后纤维软骨细胞活性的研究 被认为是同种异体移植后结果的决定因素之一。我们之前已经 开发了一种无冰玻璃化冷冻保存方法，该方法可保持良好的软骨细胞活力 动物模型和人类关节软骨。我们在这里提供的初步数据表明 半月板中的纤维软骨细胞也可以在玻璃化冷冻中存活下来，因为在冷却和冷冻过程中没有结冰 将1-3毫升的小样品加热。然而，由于结冰，无法对更大的样品进行重新加热 在复温过程中导致细胞活力丧失的核化。这项提案的创新之处在于一项新的 不存在边界对流增温应有效的局限性的复温方法 适用于体积不超过50毫升的样品。这种复温方法利用射频感应加热磁场 铁纳米粒子。我们将优化全厚度半月板纤维软骨塞的纳米温控以进行维护 细胞活力，包括基因表达，以及细胞外基质的完整性。这一目标将在#年制定 两个具体的目标是优化纳米温控变量，并使用组织塞从5ml扩大到30ml 半月板完好无损。半月板将被玻璃化保存1-4周，然后纳米臂半月板细胞存活， 化学和生物材料特性将与未经处理的新鲜对照组织进行比较。纳米武器 提供最好的细胞保存的条件，如果有任何生物材料的变化，最小的变化将被选择 用于进一步的活体研究和在未来的第二阶段SBIR中移植到人半月板上 申请。