Ice-free vitrification and nanowarming of large osteochondral grafts for transplantation

用于移植的大型骨软骨移植物的无冰玻璃化和纳米加温

• 批准号: 9918800
• 负责人: Kelvin G.M. Brockbank
• 金额: $ 83.84万
• 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 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9918800
• 关键词: Allogenic; Allografting; Animal Model; Beavers; Biocompatible Materials; Cardiovascular system; Cartilage; Cell Survival; Cells; Chemistry; Chondrocytes; Clinical; Clinical Research; Contralateral; Convection; Coupling; Cryopreservation; Custom; Data; Defect; Degenerative polyarthritis; Development; Effectiveness; Ensure; Evaluation; Extracellular Matrix; Family suidae; Farming environment; Food Processing; Formulation; Freedom; Freezing; Frequencies; Future; Heating; Histology; Human; IACUC; Ice; Implant; In Vitro; Investigation; Iron; Lead; Magnetism; Maintenance; Metabolic; Methods; Miniature Swine; Modeling; Outcome; Patients; Permeability; Phase; Plants; Property; Recovery; Research; Review Literature; Rewarming; Sample Size; Sampling; Small Business Innovation Research Grant; Surface; Surveys; Technology; Testing; Thick; Time; Tissue Engineering; Tissue Viability; Tissues; Transplantation; Trauma; articular cartilage; attenuation; biomaterial compatibility; cartilage cell; commercialization; implantation; improved; in vivo; in vivo evaluation; innovation; nanoparticle; nanowarming; osteochondral tissue; particle; phase 1 study; preclinical study; preservation; radio frequency; reconstruction; repaired; response; success; translation to humans; transplant model

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 had not been possible to rewarm larger samples due to ice nucleation during rewarming that results in loss of chondrocyte viability prior to our Phase I studies. 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. In Phase I we demonstrated the effectiveness of ice-free vitrification combined with nanowarming for large full thickness osteochondral tissues in 50 mL volumes in 80 seconds with maintenance of both chondrocyte viability and extracellular matrix integrity. In Phase II we propose development of a porcine model, further optimization of ice-free vitrification and nanowarming and in vivo evaluation in a porcine model in three specific aims. Nanowarmed chondrocyte viability, chemistry, and biomaterial properties will be compared with untreated fresh control tissues both before and after transplant. The nanowarming conditions that provides the best preservation of chondrocytes with minimal if any cartilage biomaterial changes will be selected for further investigation and translation to human cartilage in a subsequent Phase IIb 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，不可能重新加热较大的样品。 导致在我们的I期研究之前，软骨细胞生存能力丧失。该提案中的创新与 没有边界对流变暖的局限性的新的重新加热方法 对于最大50毫升的样品有效。这种重新加华的方法利用了射频引起的加热 磁铁纳米颗粒。在第一阶段，我们证明了无冰玻璃化的有效性 使用纳米武器，可在80秒内以50 mL的体积为50 ml的大厚度骨软骨组织 维持软骨细胞生存能力和细胞外基质完整性。在第二阶段，我们提出了 开发猪模型，进一步优化无冰的玻璃化和纳米臂和体内 在猪模型中评估三个特定目标。纳米臂的软骨细胞生存能力，化学和 在移植前后，将将生物材料与未处理的新鲜对照组织进行比较。 纳米武器条件可提供最佳的软骨细胞，如果有任何软骨 将选择生物材料变化，以进一步研究并转化为人类软骨 随后的IIB SBIR应用程序。