Although it is relatively straightforward to cryopreserve living isolated chondrocytes, at the present time there is no satisfactory method to preserve surgical grafts between the time of procurement or manufacture and actual use. In earlier papers we have established that the cryoprotectants dimethyl sulphoxide or propylene glycol do penetrate into this tissue very rapidly. Chondrocytes are not unusually susceptible to osmotic stress; in fact they appear to be particularly resistant. It appears that damage is associated with the formation of ice per se, even at cooling rates that are optimal for the cryopreservation of isolated chondrocytes. We then showed that current methods of cartilage cryopreservation involve the nucleation and growth of ice crystals within the chondrons rather than ice being restricted to the surrounding acellular matrix. This finding established the need to avoid the crystallization of ice-in other words, vitrification. Song and his colleagues have published a vitrification method that is based on the use of one of Fahy's vitrification formulations. We confirmed the effectiveness of this method but found it to be very dependent on ultra rapid warming. However, we were able to develop a 'liquidus-tracking' method that completely avoids the crystallization of ice and does not require rapid warming. The ability of cartilage preserved in this way to incorporate sulphate into newly synthesized glycosaminoglycans (GAGs) approached 70% of that of fresh control cartilage. In this method the rates of cooling and warming can be very low, which is essential for any method that is to be used in Tissue Banks to process the bulky grafts that are required by orthopaedic surgeons. Work is continuing to refine this method for Tissue Bank use.

中文翻译：

---

关节软骨的冷冻保存。第 3 部分：液相线跟踪方法

虽然冷冻保存活的分离的软骨细胞相对简单，但目前还没有令人满意的方法来保存手术移植物在采购或制造与实际使用之间的时间。在早期的论文中，我们已经确定冷冻保护剂二甲基亚砜或丙二醇确实非常迅速地渗透到该组织中。软骨细胞对渗透压并不异常敏感。事实上，他们似乎特别有抵抗力。损伤似乎与冰的形成本身有关，即使在对分离的软骨细胞进行冷冻保存的最佳冷却速率下也是如此。然后我们表明，目前的软骨冷冻保存方法涉及软骨内冰晶的成核和生长，而不是将冰限制在周围的无细胞基质中。这一发现确立了避免冰结晶的必要性——换句话说，玻璃化。Song 和他的同事发表了一种基于使用 Fahy 的一种玻璃化配方的玻璃化方法。我们证实了这种方法的有效性，但发现它非常依赖于超快速变暖。然而，我们能够开发出一种“液相线跟踪”方法，它完全避免了冰的结晶并且不需要快速升温。以这种方式保存的软骨将硫酸盐结合到新合成的糖胺聚糖 (GAG) 中的能力接近新鲜对照软骨的 70%。在这种方法中，降温和升温的速度可能非常低，这对于在组织库中使用的任何方法来处理整形外科医生所需的庞大移植物都是必不可少的。正在继续改进此方法以供组织库使用。