Hahn Joshua, Laouar Leila, Elliott Janet A W, Korbutt Gregory S, Jomha Nadr M
Department of Surgery, University of Alberta Hospital, 2D2.28 WMC, 8440-112St, Edmonton, Alberta T6G 2B7, Canada.
Department of Chemical and Materials Engineering, University of Alberta, Edmonton, Alberta T6G 1H9, Canada; Department of Laboratory Medicine and Pathology, University of Alberta, Edmonton, Alberta T6G 2B7, Canada.
Cryobiology. 2017 Apr;75:68-74. doi: 10.1016/j.cryobiol.2017.02.002. Epub 2017 Feb 10.
High concentrations of cryoprotective agents are required for cryopreservation techniques such as vitrification. Glycerol is a common cryoprotective agent used in cryopreservation protocols but this agent is toxic at high concentrations. This work is an attempt to mitigate the toxic effects of high concentrations of glycerol on intact chondrocytes in human knee articular cartilage from total knee arthroplasty patients by simultaneous exposure to glycerol and a variety of additive compounds. The resulting cell viability in the cartilage samples as measured by membrane integrity staining showed that, in at least one concentration or in combination, all of the tested additive compounds (tetramethylpyrazine, ascorbic acid, chondroitin sulphate, glucosamine sulphate) were able to reduce the deleterious effects of glycerol exposure when examination of membrane integrity took place on a delayed time frame. The use of additive compounds to reduce cryoprotectant toxicity in articular cartilage may help improve cell recovery after cryopreservation.
诸如玻璃化冷冻等冷冻保存技术需要高浓度的冷冻保护剂。甘油是冷冻保存方案中常用的冷冻保护剂,但该试剂在高浓度时具有毒性。这项研究旨在通过同时将人全膝关节置换术患者的膝关节软骨中的完整软骨细胞暴露于甘油和多种添加剂化合物,来减轻高浓度甘油对其的毒性作用。通过膜完整性染色测量软骨样本中的细胞活力,结果表明,在至少一种浓度或组合中,当在延迟时间框架内检查膜完整性时,所有测试的添加剂化合物(川芎嗪、抗坏血酸、硫酸软骨素、硫酸氨基葡萄糖)都能够降低甘油暴露的有害影响。使用添加剂化合物来降低关节软骨中冷冻保护剂的毒性可能有助于改善冷冻保存后的细胞恢复。