UNASSIGNED: CPAs were tested on bovine nucleus pulposus cells to determine the least cytotoxic solution. Capitalizing on our CPAs Computed Tomography (CT) contrast enhancement, we imaged and quantified saturation time in intact bovine IVDs under different conditions in a bioreactor. Finally, the entire protocol was tested, including 1 week of frozen storage, to confirm tissue viability in multiple IVD regions after thawing.
UNASSIGNED: Results showed cryopreserving medium containing dimethyl sulfoxide and ethylene glycol gave over 7.5 h before cytotoxicity. While non-loaded IVDs required over 3 days to fully saturate, a dynamic loading protocol followed by CPA addition and free-swelling decreased saturation time to <5 h. After cryopreserving IVDs for 1 week with the optimized CPA and permeation method, all IVD regions had 85% cell viability, not significantly different from fresh unfrozen controls.
UNASSIGNED: This study created a novel solution to a roadblock in IVD research and development. Using post-compression swelling CPA can be delivered to an intact IVD over 20× more quickly than previous methods, enabling cryopreservation of the IVD with no detectable loss in cell viability.
■在牛髓核细胞上测试CPAs以确定细胞毒性最小的溶液。利用我们的注册会计师计算机断层扫描(CT)对比增强,我们在生物反应器中的不同条件下对完整的牛IVD的饱和时间进行了成像和定量。最后,整个协议都经过了测试,包括一周的冷冻储存,以确认解冻后多个IVD区域的组织活力。
■结果显示含有二甲基亚砜和乙二醇的冷冻保存培养基在细胞毒性之前超过7.5小时。未加载的IVD需要超过3天才能完全饱和,添加CPA和自由溶胀后的动态加载方案将饱和时间降低至<5小时。使用优化的CPA和渗透方法冷冻保存IVD1周后,所有IVD区域都有85%的细胞活力,与新鲜的未冷冻对照没有显着差异。
■这项研究为IVD研究和开发中的障碍创造了一种新颖的解决方案。使用压缩后肿胀CPA可以比以前的方法更快地递送到完整的IVD超过20倍。能够冷冻保存IVD,而细胞活力没有可检测到的损失。