Ice Free Vitrification and nanowarming of large cartilage samples for transplantation

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 导致软骨细胞活力丧失。这项提案的创新涉及一种新的复温方法， 没有边界对流变暖的限制，这种限制对50毫升以下的样品有效 在音量上。这种复温方法利用射频感应加热磁性铁纳米颗粒。我们 将优化全层骨吸收存储的纳米温控，以维持软骨细胞的活力 和细胞外基质的完整性。这一目标将在两个具体目标中发展，以优化纳米武器 变量和规模从5毫升到30-50毫升。纳米级软骨细胞的活性、化学和 生物材料的性能将与未经处理的新鲜对照组织进行比较。纳米武器的条件 提供最好的软骨细胞保存，即使有软骨生物材料的变化也是最小的 选择在体内进行进一步研究并在随后的第二阶段SBIR中移植到人软骨中 申请。

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.