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：有限的新鲜活体供体椎间盘移植物可获得性降低了尺寸匹配效率， 同种异体骨移植，限制手术可及性和患者结局。当前保存 保留细胞活力的方法是不够的。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。我们先前已经 开发了一种冷冻保存的无冰玻璃化方法，该方法在体外培养中保持了良好的软骨细胞活力， 动物模型和人关节软骨。我们在此提供的初步数据表明， 由于在冷却过程中没有冰形成， 加热1- 3 mL小样品。然而，由于冰的存在， 在复温过程中的成核，导致细胞活力的丧失。本提案的创新之处在于， 一种没有边界对流加热限制的复温方法，应该是有效的 对于体积高达50 mL的样品。这种复温方法利用射频感应加热的磁性材料， 铁纳米颗粒我们将优化全厚度椎间盘纤维软骨塞的修复，以进行维护 包括基因表达和细胞外基质完整性。这一目标将在 两个具体目标是优化包装变量，并从使用组织塞的5 mL扩大到30 mL， 完整的包皮半月板将被玻璃化并储存1-4周，然后检测半月板细胞活力， 化学性质和生物材料性质将与未处理的新鲜对照组织进行比较。第1001章武装 将选择提供最佳细胞保存且生物材料变化最小（如果有）的条件 为了在体内进行进一步研究，并在随后的未来II期SBIR中转化为人源性大肠杆菌， 应用程序.