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.

冷藏同种异体骨软骨表面置换术在临床上的应用 创伤和骨关节炎所致的关节软骨表面损伤。移植软骨细胞活性的研究 关节软骨被认为是同种异体骨软骨移植结果的决定因素之一。 移植。我们之前已经开发出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良好的软骨细胞活性。软骨细胞 在1-3毫升样品的冷却和升温过程中，由于没有结冰，玻璃化保存下来。 然而，由于复温过程中的冰核作用，无法对较大的样品进行复温 结果在我们的第一阶段研究之前，软骨细胞的活性丧失。这项提案的创新之处在于 新的复温方法没有边界对流增温的限制，应该是 适用于体积不超过50毫升的样品。这种复温方法利用射频感应加热 磁性铁纳米粒子。在第一阶段，我们演示了无冰玻璃化联合冷冻的有效性。 在80秒内对体积为50毫升的大片全层骨软骨组织进行纳米温控 维持软骨细胞的活力和细胞外基质的完整性。在第二阶段，我们建议 猪模型的建立、无冰玻璃化冷冻和纳米温控技术的进一步优化及体内应用 在猪模型中对三个特定目标进行评估。纳米级软骨细胞的活性、化学和 移植前后的生物材料特性将与未经处理的新鲜对照组织进行比较。 以最少的软骨提供最好的软骨细胞保存的纳米温控条件 将选择生物材料变化进行进一步研究，并将其移植到人类软骨中 后续阶段IIb SBIR申请。