Ice Free Vitrification and nanowarming of large cartilage samples for transplantation

用于移植的大型软骨样本的无冰玻璃化和纳米加温

基本信息

批准号：
9473828
负责人：
Kelvin G.M. Brockbank
金额：
$ 15万
依托单位：
TISSUE TESTING TECHNOLOGIES, LLC
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-07 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9473828
关键词：
Allogenic Allografting Animal Model Animals Beavers Biocompatible Materials Biological Assay Biological Preservation Caliber Cardiovascular system Cartilage Cell Culture Techniques Cell Survival Cells Chemistry Chondrocytes Clinical Clinical Research Complex Convection Coupling Cryopreservation Data Defect Degenerative polyarthritis Effectiveness Ensure Evaluation Extracellular Matrix Family suidae Food Processing Formulation Freedom Freezing Frequencies Future Harvest Heating Human IACUC Ice In Vitro Investigation Iron Life Magnetism Maintenance Metabolic Methods Outcome Patients Permeability Pharmacopoeias Phase Plants Plug-in Property Research Review Literature Rewarming Sample Size Sampling Small Business Innovation Research Grant Surface Surveys Technology Testing Thick Time Tissue Engineering Tissue Sample Tissue Viability Tissues Transplantation Trauma Trypan Blue United States articular cartilage attenuation biomaterial compatibility bone cartilage allograft cartilage repair commercialization implantation in vivo innovation nanoparticle osteochondral tissue particle radio frequency reconstruction repaired response translation to humans

项目摘要

Resurfacing of articular cartilage with cold stored osteochondral allografts is employed clinically for repair of trauma and osteoarthritis-induced articular cartilage surface damage. Chondrocyte viability of transplanted articular cartilage is accepted as one of the determinants of outcome following osteochondral allograft transplantation. We have previously developed an ice-free vitrification method of cryopreservation that maintains excellent chondrocyte viability in animal model and human articular cartilage. The chondrocytes survive vitrification due to the absence of ice formation during cooling and warming of 1-3mL samples. However, it has not been possible to rewarm larger samples due to ice nucleation during rewarming that results in loss of chondrocyte 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 osteocartilage storage for maintenance of both chondrocyte viability and extracellular matrix integrity. This objective will be developed in two specific aims to optimize nanowarming variables and scaleup from 5mL to 30-50mL volumes. Nanowarmed chondrocyte viability, chemistry, and biomaterial properties will be compared with untreated fresh control tissue. The nanowarming conditions that provides the best preservation of chondrocytes with minimal if any cartilage biomaterial changes will be selected for further investigation in vivo and translation to human cartilage in a subsequent Phase II SBIR application.

临床上采用冷保存骨软骨同种异体移植物进行关节软骨表面修复创伤和骨关节炎引起的关节软骨表面损伤。移植软骨细胞的活力关节软骨被认为是骨软骨同种异体移植后结果的决定因素之一移植。我们之前开发了一种无冰玻璃化冷冻保存方法在动物模型和人类关节软骨中保持优异的软骨细胞活力。软骨细胞由于在 1-3mL 样品冷却和升温过程中不形成冰，因此能够在玻璃化过程中幸存。然而，由于复温过程中冰核的存在，无法对较大的样品进行复温导致软骨细胞活力丧失。该提案的创新涉及一种新的复温方法，该方法没有边界对流升温的限制，该限制对 50mL 以下的样品有效在体积上。这种复温方法利用磁性铁纳米粒子的射频感应加热。我们将优化全层骨软骨储存的纳米变暖，以维持软骨细胞的活力和细胞外基质的完整性。该目标将分为两个具体目标来优化纳米变暖变量和从 5mL 到 30-50mL 体积的放大。纳米加热软骨细胞活力、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。纳米变暖条件提供软骨细胞的最佳保存，并且软骨生物材料的变化最小（如果有的话）选择在随后的 II 期 SBIR 中进行进一步的体内研究并转化为人类软骨应用。