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首页> 外文会议>Symposium on Advanced Biomaterials―Characterization, Tissue Engineering and Complexity, Nov 26-29, 2001, Boston, Massachusetts, U.S.A. >A Quartz Crystal Microbalance Cell Biosensor: Detecting Nocodazole Dependent Microtubule Disruption Dynamics In Living Cells
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A Quartz Crystal Microbalance Cell Biosensor: Detecting Nocodazole Dependent Microtubule Disruption Dynamics In Living Cells

机译：石英晶体微天平细胞生物传感器：检测活细胞中依赖诺考达唑的微管破坏动力学

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摘要

A Quartz Crystal Microbalance (QCM) was used to create a biosensor utilizing living adherent endothelial cells (ECs) as the biological sensing element. This EC QCM biosensor detected the effect of varying concentrations of nocodazole, a microtubule binding and disrupting drug, on the adherent cells as they altered the underlying QCM device state frequency, Δf, and motional Resistance, ΔR, shift values. Over the dose range 0.11-15 μM nocodazole, the Δf shift values decreased significantly in magnitude in a dose dependent fashion over a 5-6 hr incubation period following drug addition to a limiting value, with a 900 nM midpoint. This effect is consistent with nocodazole's known dose dependent effect on the disruption of microtubules. At all drug concentrations, the relative Δf decrease with time was found to be very similar and well fit by a single exponential decay equation. For all nocodazole doses, t_(0.5) was invariant, averaging t_(0.5) = 0.83 +- 0.14 hr. These data demonstrate that a single dynamic sensing system within the cell, the microtubules, responds to the addition of nocodazole and its response can be quantified by the biosensor. These results indicate that the EC QCM biosensor can be used to detect EC cytoskeletal alterations and dynamics and may be a valuable screening method for all classes of biologically active drugs or biological macromolecules that affect cytoskeleton perturbations or cellular attachment.
机译：石英晶体微量天平（QCM）用于创建生物传感器，利用活的粘附内皮细胞（EC）作为生物传感元件。这款EC QCM生物传感器检测到不同浓度的诺考达唑（一种微管结合和破坏药物）对贴壁细胞的影响，因为它们改变了潜在的QCM设备状态频率Δf和运动阻力ΔR位移值。在0.11-15μM诺考达唑的剂量范围内，药物添加后的5-6小时孵育期内，Δf位移值以剂量依赖性方式显着降低，幅度极限为900 nM。该作用与诺考达唑对微管破坏的已知剂量依赖性作用一致。在所有药物浓度下，相对Δf随时间的下降都非常相似，并且可以通过单个指数衰减方程很好地拟合。对于所有诺考达唑剂量，t_（0.5）不变，平均t_（0.5）= 0.83±0.14 hr。这些数据表明，细胞内的单个动态传感系统（微管）对诺考达唑的添加有反应，并且其反应可以通过生物传感器进行量化。这些结果表明，EC QCM生物传感器可用于检测EC细胞骨架的变化和动力学，并且可能是影响细胞骨架扰动或细胞附着的所有类型的生物活性药物或生物大分子的有价值的筛选方法。

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