Open access to novel dual flow chamber technology for in vitro cell mechanotransduction, toxicity and pharamacokinetic studies.
Open access to novel dual flow chamber technology for in vitro cell mechanotransduction, toxicity and pharamacokinetic studies.
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DOI：
10.1186/1475-925x-6-46
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发表时间：
2007-12-04
影响因子：
3.9
通讯作者：
Knothe Tate ML
中科院分区：
文献类型：
--
作者：
Anderson EJ;Knothe Tate ML
研究人员开发机械转导实验模型的主要障碍是实验变量的控制，特别是在细胞水平上的机械力的传递。先前对现有技术商业灌注室的评估表明，用于向细胞施加限定的机械刺激的流动状态控制不佳，并且即使目标应力状态相当，也不能比较使用不同室的研究的数据。本研究提供了一种新颖的腔室设计，以提供基于生理学的流态、实验变量控制的改进以及与商业腔室相比的易用性。这种新颖的设计通过五种垫圈设计以及单流和双流状态实现了受控应力。垫片几何形状内施加的剪应力得到很好的控制。10 × 21 mm通用垫圈（垫圈I，设计用于在进行结果测量的灭菌室中心施加恒定大小的剪切应力）整个面积的50%暴露于目标应力。在腔室中心的8 mm直径圆形区域（进行结局测量）中，超过92%的区域暴露于目标应力（± 2.5%）。此外，其它垫圈几何形状提供随离腔室入口的距离而变化的特定应力梯度。对新型腔室原型的台式测试表明，与其他商业腔室相比，在易用性和性能方面都有改进。腔室的设计消除了由于泄漏和气泡而导致的流量偏差，并允许实际流量剖面更好地符合计算模型中的预测。新型流动室设计在细胞单层表面提供了可预测且明确定义的机械力，显示出优于先前测试的商业室的改进。施加应力的可预测性提高了实验的可重复性以及研究间比较的准确性。仔细控制细胞上的应力对于有效模拟体内情况至关重要。总的来说，改进的灌注流动室提供了所需的分辨率、标准化和类似于体内条件的体外模型，从而朝着在研究中更大的用途迈出了一步，并有机会进入诊断和治疗市场。
A major stumbling block for researchers developing experimental models of mechanotransduction is the control of experimental variables, in particular the transmission of the mechanical forces at the cellular level. A previous evaluation of state of the art commercial perfusion chambers showed that flow regimes, applied to impart a defined mechanical stimulus to cells, are poorly controlled and that data from studies in which different chambers are utilized can not be compared, even if the target stress regimes are comparable. This study provides a novel chamber design to provide both physiologically-based flow regimes, improvements in control of experimental variables, as well as ease of use compared to commercial chambers. This novel design achieves controlled stresses through five gasket designs and both single- and dual-flow regimes. The imparted shear stress within the gasket geometry is well controlled. Fifty percent of the entire area of the 10 × 21 mm universal gasket (Gasket I, designed to impart constant magnitude shear stresses in the center of the chamber where outcome measures are taken), is exposed to target stresses. In the 8 mm diameter circular area at the center of the chamber (where outcome measures are made), over 92% of the area is exposed to the target stress (± 2.5%). In addition, other gasket geometries provide specific gradients of stress that vary with distance from the chamber inlet. Bench-top testing of the novel chamber prototype shows improvements, in the ease of use as well as in performance, compared to the other commercial chambers. The design of the chamber eliminates flow deviations due to leakage and bubbles and allows actual flow profiles to better conform with those predicted in computational models. The novel flow chamber design provides predictable and well defined mechanical forces at the surface of a cell monolayer, showing improvement over previously tested commercial chambers. The predictability of the imparted stress improves both experiment repeatability as well as the accuracy of inter-study comparisons. Carefully controlling the stresses on cells is critical in effectively mimicking in vivo situations. Overall, the improved perfusion flow chamber provides the needed resolution, standardization and in vitro model analogous to in vivo conditions to make the step towards greater use in research and the opportunity to enter the diagnostic and therapeutic market.