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

基本信息

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
项目类别：
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

项目摘要

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：新鲜可行的供体半月板移植物的供应有限，降低了尺寸匹配效率半月板同种异体移植，限制了手术的可及性和患者的预后。当前保存保留半月板活力的方法是不够的。移植半月板的纤维软骨细胞活力被认为是同种异体移植后结果的决定因素之一。我们之前有过开发了一种冷冻保存的无冰玻璃化方法，可在动物模型和人类关节软骨。我们在此提供初步数据表明由于在冷却和冷冻过程中没有冰形成，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。加热 1-3mL 小样品。然而，由于冰的存在，无法对较大的样品进行重新加热复温期间成核，导致细胞活力丧失。该提案的创新点涉及一种新的没有边界对流升温限制的复温方法应该是有效的适用于体积高达 50mL 的样品。这种复温方法利用磁力的射频感应加热铁纳米颗粒。我们将优化全厚度半月板纤维软骨塞的纳米加热以进行维护细胞活力，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纳米变暖变量，并从使用组织塞的 5mL 扩大到使用组织塞的 30mL 完整的半月板。半月板将被玻璃化并储存 1-4 周，然后纳米加热半月板细胞活力，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。纳米变暖将选择提供最佳细胞保存且生物材料变化最小的条件在随后的未来 II 期 SBIR 中进一步进行体内研究并转化为人类半月板应用。