Michelle D. Kofron, Natalie C. Opsitnick, Mohamed A. Attawia, Cato T. Laurencin
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引用次数: 33
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Abstract
The large-scale clinical use of tissue engineered constructs will require provisions for its mass availability and accessibility. Therefore, it is imperative to understand the effects of low temperature (–196°C) on the tissue engineered biological system. Initial studies used samples of the osteoblast-like cell line (SaOS-2) adhered to a two-dimensional poly(lactide-co -glycolide) thin film (2D-PLAGA) or a three-dimensional poly(lactide-co -glycolide) sintered microsphere matrix (3D-PLAGA) designed for bone tissue engineering. Experimental samples were tested for their ability to maintain cell viability, following low temperature banking for one week, in solutions of the penetrating cryoprotective agents, dimethylsulfoxide (DMSO), ethylene glycol, and glycerol. Results indicated the DMSO solution yielded the greatest percent cell survival for SaOS-2 cells adhered to both the 2D- and 3D-PLAGA scaffolds; therefore, DMSO was used to cryopreserve mineralizing primary rabbit osteoblasts cells adhered to 2D-PLAGA matrices for 35 days. Results indicated retention of the extracellular matrix architecture as no statistically significant difference in the pre- and post-thaw mineralized structures was measured. Percent cell viability of the mineralized constructs following low temperature storage was approximately 50%. These are the first studies to address the issue of preservation techniques for tissue engineered constructs. The ability to successfully cryopreserve mineralized tissue engineered matrices for bone may offer an unlimited and readily available source of bone-like materials for orthopaedic applications. © 2003 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved.
骨组织工程构建物的低温保存
组织工程结构的大规模临床应用将需要对其大量可用性和可及性作出规定。因此,了解低温(-196°C)对组织工程生物系统的影响势在必行。最初的研究将成骨细胞样细胞系(SaOS-2)的样品粘附在用于骨组织工程的二维聚(乳酸-羟基乙酸酯)薄膜(2D-PLAGA)或三维聚(乳酸-羟基乙酸酯)烧结微球基质(3D-PLAGA)上。实验样品在低温保存一周后,在穿透性冷冻保护剂二甲基亚砜(DMSO)、乙二醇和甘油的溶液中测试其维持细胞活力的能力。结果表明,DMSO溶液对粘附在2D和3D-PLAGA支架上的SaOS-2细胞的细胞存活率最高;因此,使用DMSO将矿化兔原代成骨细胞粘附在2D-PLAGA基质上冷冻保存35天。结果表明,细胞外基质结构的保留在解冻前和解冻后的矿化结构中没有统计学上的显著差异。低温储存后,矿化构建体的细胞存活率约为50%。这些是解决组织工程构建的保存技术问题的第一个研究。成功低温保存矿化组织工程骨基质的能力可能为骨科应用提供无限且容易获得的骨样材料来源。©2003骨科研究学会。Elsevier Science Ltd.出版。版权所有。
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