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

• 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
• 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

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.

用冷储存的骨软骨同种异体移植关节软骨在临床上用于修复 创伤和骨关节炎引起的关节软骨表面损伤。移植的软骨细胞生存力 关节软骨被接受为骨软骨后的结果的决定因素之一 移植。我们以前已经开发了一种无冰的玻璃化方法的冷冻保存方法 在动物模型和人类关节软骨中保持出色的软骨细胞活力。软骨细胞 由于在1-3ml样品的冷却和变暖过程中，由于没有冰的形成而生存的玻璃化。 但是，由于冰核在重新加热过程中，不可能重新加热大型样品。 导致软骨细胞生存能力丧失。该提案中的创新与一种新的重新加热方法有关 没有边界对流变暖的局限 数量。这种重新传输方法利用了射频诱导的磁铁纳米颗粒的加热。我们 将优化全厚度骨软骨存储的纳米武器，以维持两种软骨细胞的生存能力 和细胞外基质完整性。该目标将以两个特定的目的开发，以优化纳米武器 变量和比例尺从5ml到30-50ml卷。纳米臂的软骨细胞生存能力，化学和 将将生物材料与未处理的新鲜对照组织进行比较。纳米武器条件 如果有任何软骨生物材料变化，则提供最佳的软骨细胞保存 在随后的II期SBIR中被选为体内进一步研究，并将其转换为人软骨 应用。