Cryoprotective agent toxicity interactions in human articular chondrocytes
Cryoprotective agent toxicity interactions in human articular chondrocytes
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DOI：
10.1016/j.cryobiol.2012.01.006
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发表时间：
2012-06-01
期刊：
影响因子：
2.7
通讯作者：
Jomha, N. M.
中科院分区：
文献类型：
--
作者：
Almansoori, K. A.;Prasad, V.;Jomha, N. M.
工作背景：玻璃化是一种冷冻保存方法，通过该方法，可以在低温下使用高浓度（通常>= 6.0 M）的冷冻保护剂（CPA）来保存细胞和组织，以限制在冷却过程中可能形成的冰晶的有害影响。然而，在这些浓度下，CPA具有显著的细胞毒性，了解其毒性特征和相互作用是重要的。因此，评价了单CPA和多CPA溶液对软骨细胞的直接和间接毒性。从人关节软骨样品中分离软骨细胞，并暴露于五种常见CPA的各种单CPA和多CPA溶液中（二甲基亚砜（DMSO），乙二醇（EG），丙二醇（PG），甘油（戈伊）和甲酰胺（Fm））在6.0和8.1M浓度下在0 ℃下处理30分钟。结果：多组CPA溶液的毒性显著低于单组CPA溶液（P <0.01），而单组CPA溶液的毒性显著低于单组CPA溶液（P < 0.01）。CPA之间的间接毒性效应可以用回归分析来量化。用四-CPA组合溶液DMSO-EG-Gy-Fm获得约40%的细胞存活率。在多个CPA组合，PG表现出最大程度的毒性和它的存在内的组合解决方案否定了使用多个较低浓度CPAs.Conclusions的任何好处：多个CPA解决方案的细胞毒性比单CPA解决方案相同的总浓度。PG是毒性最大的CPA组合使用时。用6.0 M DMSO-EG-Gy-Fm组合溶液获得最高的软骨细胞存活率。(C)2012 Elsevier Inc. All rights reserved.
Background: Vitrification is a method of cryopreservation by which cells and tissues can be preserved at low temperatures using cryoprotective agents (CPAs) at high concentrations (typically >= 6.0 M) to limit the harmful effects of ice crystals that can form during cooling processes. However, at these concentrations CPAs are significantly cytotoxic and an understanding of their toxicity characteristics and interactions is important. Therefore, single-CPA and multiple-CPA solutions were evaluated for their direct and indirect toxicities on chondrocytes.Methods: Chondrocytes were isolated from human articular cartilage samples and exposed to various single-CPA and multiple-CPA solutions of five common CPAs (dimethyl sulfoxide (DMSO), ethylene glycol (EG), propylene glycol (PG), glycerol (Gy) and formamide (Fm)) at both 6.0 and 8.1 M concentrations at 0 degrees C for 30 min. Chondrocyte survival was determined using a fluorescent cell membrane integrity assay. The data obtained was statistically analyzed and regression coefficients were used to represent the indirect toxicity effect which a specific combination of CPAs exerted on the final solution's toxicity.Results: Multiple-CPA solutions were significantly less toxic than single-CPA solutions (P < 0.01). The indirect toxicity effects between CPAs were quantifiable using regression analysis. Cell survival rates of approximately 40% were obtained with the four-CPA combination solution DMSO-EG-Gy-Fm. In the multiple-CPA combinations, PG demonstrated the greatest degree of toxicity and its presence within a combination solution negated any benefits of using multiple lower concentration CPAs.Conclusions: Multiple-CPA solutions are less cytotoxic than single-CPA solutions of the same total concentration. PG was the most toxic CPA when used in combinations. The highest chondrocyte survival rates were obtained with the 6.0 M DMSO-EG-Gy-Fm combination solution. (C) 2012 Elsevier Inc. All rights reserved.