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用于细胞构建体的振荡压缩装置的设计与应用

Design and application of an oscillatory compression device for cell constructs.

作者信息

Cassino Theresa R, Anderson Roger, Love Brian J, Huckle William R, Seamans Diane K, Forsten-Williams Kimberly

机构信息

Department of Chemical Engineering, Virginia Polytechnic Institute and State University, Virginia, USA.

出版信息

Biotechnol Bioeng. 2007 Sep 1;98(1):211-20. doi: 10.1002/bit.21422.

Abstract

Mechanical compression has been shown to impact cell activity; however a need for a single device to perform a broader range of parametric studies exists. We have developed an oscillatory displacement controlled device to uniaxially strain cell constructs under both static and dynamic compression and used this device to investigate gene expression in cell constructs. The device has a wide stroke (0.25-4 mm) and frequency range (0.1-3 Hz) and several loading waveforms are possible. Alginate cellular constructs with embedded equine chondrocytes were tested and viability was maintained for the 24 h test period. Off-line mechanical testing is described and a modulus value of 18.2 +/- 1.3 kPa found for alginate disks which indicates the level of stress achieved with this deformation profile. Static (15% strain) and dynamic (15% strain, 1 Hz, triangle waveform) testing of chondrocyte constructs was performed and static compression showed significantly higher collagen II expression than dynamic using quantitative RT-PCR. In contrast, differences in matrix metalloproteinase-3 (MMP-3) expression were statistically insignificant. These studies indicate the utility of our device for studying cell activity in response to compression and suggest further studies regarding how the load and strain spectrum impact chondrocyte activity.

摘要

机械压缩已被证明会影响细胞活性;然而,需要一种能够进行更广泛参数研究的单一装置。我们开发了一种振荡位移控制装置,用于在静态和动态压缩下对细胞构建体进行单轴拉伸,并使用该装置研究细胞构建体中的基因表达。该装置具有宽行程(0.25 - 4毫米)和频率范围(0.1 - 3赫兹),并且有几种加载波形可供选择。对嵌入马软骨细胞的藻酸盐细胞构建体进行了测试,在24小时的测试期内保持了活力。描述了离线机械测试,并发现藻酸盐圆盘的模量值为18.2±1.3千帕,这表明了这种变形曲线所达到的应力水平。对软骨细胞构建体进行了静态(15%应变)和动态(15%应变,1赫兹,三角波形)测试,使用定量逆转录聚合酶链反应(RT-PCR)发现,静态压缩显示出比动态压缩显著更高的胶原蛋白II表达。相比之下,基质金属蛋白酶-3(MMP-3)表达的差异在统计学上不显著。这些研究表明我们的装置在研究细胞对压缩的反应活性方面的实用性,并建议进一步研究载荷和应变谱如何影响软骨细胞活性。

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