Intramatrix events during cryopreservation of porcine articular cartilage using rapid cooling
Intramatrix events during cryopreservation of porcine articular cartilage using rapid cooling
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DOI：
10.1016/s0736-0266(03)00158-x
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发表时间：
2004-01-01
影响因子：
2.8
通讯作者：
McGann, LE
中科院分区：
文献类型：
--
作者：
Jomha, NM;Anoop, PC;McGann, LE
如果能够保持细胞和基质的完整性，关节软骨的冷冻保存可能会改善长期的移植效果。本研究检测了在不同浓度的二甲基亚砜(DMSO)(1、3、5、6和7M)快速冷却技术过程中，完整猪关节软骨中发生的基质内事件。将热电偶插入溶液和软骨基质中，以记录快速冷却期间的温度。此外，还对冷冻替代样品进行了扫描电子显微镜检查，并对基质中代表冰的区域进行了定量评估。研究结果表明，低浓度的DMSO导致了基质与周围溶液之间的最大温度梯度，该温度梯度发生在低温保护剂溶液的冰点附近。当DMSO浓度较高时，峰值温度梯度出现在玻璃化转变温度附近。温度测量表明，在较低的DMSO浓度下，基质中形成了大量的冰。在导致外部溶液玻璃化的较高DMSO浓度下，有证据表明基质中有一些冰。扫描电子显微镜显示，在较低的DMSO浓度(1和5M)下(可能是由于结冰)(P<；0.02)，较低的DMSO浓度显示出更多的基质破坏，而6M的DMSO浓度显示最小的基质破坏。用快速降温技术和高浓度的二甲基亚砜冷冻保存关节软骨，可导致基质部分玻璃化，并显著减少基质破坏。看来，成功地冷冻保存关节软骨的活性和功能将需要高浓度的冷冻保护剂和快速冷却。(C)2003年骨科研究会。爱思唯尔有限公司出版。保留所有权利。
Cryopreservation of articular cartilage may improve long-term transplantation results if cell and matrix integrity can be maintained. This study examined intramatrix events in intact porcine articular cartilage that occurred during a rapid-cooling technique with various concentrations of dimethyl sulfoxide (DMSO) (1, 3, 5, 6 and 7 M). Thermocouples were inserted into the solution and in the cartilage matrix to record the temperature during rapid cooling. In addition, scanning electron microscopy of freeze-substituted samples was performed and quantitatively evaluated for the areas representing ice in the matrix. The results of this study showed that low concentrations of DMSO resulted in the largest temperature gradient between the matrix and the surrounding solution, which occurred near the freezing point of the cryoprotectant solution. At higher concentrations of DMSO, the peak temperature gradient occurred near the glass transition temperature. The temperature measurements suggested that a significant amount of ice formed within the matrix at lower DMSO concentrations. At higher DMSO concentrations that resulted in vitrification of the external solution, there was evidence of some ice in the matrix. The scanning electron micrographs demonstrated significantly more matrix disruption (likely due to ice formation) (P < 0.02) in the lower DMSO concentrations (1 and 5 M) while the 6 M DMSO concentration demonstrated minimal matrix disruption. Cryopreservation of articular cartilage with a rapid-cooling technique and high concentrations of DMSO resulted in partial vitrification of the matrix and significantly less matrix disruption. It appears that successful cryopreservation of viability and function in articular cartilage will require high concentrations of cryoprotectants and rapid cooling. (C) 2003 Orthopaedic Research Society. Published by Elsevier Ltd. All rights reserved.