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.

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，不可能对较大的样品进行复温。 在我们的第一阶段研究之前导致软骨细胞活力丧失。该提案的创新点涉及 新的复温方法不存在边界对流增温的局限性，而边界对流增温应该是 对于体积高达 50mL 的样品有效。这种复温方法利用射频感应加热 磁性铁纳米颗粒。在第一阶段，我们展示了无冰玻璃化结合的有效性 80 秒内对 50 mL 体积的大型全层骨软骨组织进行纳米加热 维持软骨细胞活力和细胞外基质完整性。在第二阶段我们建议 猪模型的开发，进一步优化无冰玻璃化和纳米变暖以及体内 在猪模型中针对三个具体目标进行评估。纳米加热软骨细胞活力、化学和 将在移植前后将生物材料特性与未经处理的新鲜对照组织进行比较。 纳米变暖条件可提供软骨细胞的最佳保存，并且软骨最少（如果有的话） 将选择生物材料的变化进行进一步研究并转化为人类软骨 随后的 IIb 期 SBIR 申请。