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Automated 3-D Printed Arrays to Evaluate Genotoxic Chemistry: E-Cigarettes and Water Samples

机译:自动化的3D打印阵列以评估遗传毒性化学物质:电子烟和水样

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摘要

A novel, automated, low cost, three-dimensional (3-D) printed microfluidic array was developed to detect DNA damage from metabolites of chemicals in environmental samples. The electrochemiluminescent (ECL) detection platform incorporates layer-by-layer (LbL) assembled films of microsomal enzymes, DNA and an ECL-emitting ruthenium metallopolymer in ∼10 nm deep microwells. Liquid samples are introduced into the array, metabolized by the human enzymes, products react with DNA if possible, and DNA damage is detected by ECL with a camera. Measurements of relative DNA damage by the array assess the genotoxic potential of the samples. The array analyzes three samples simultaneously in 5 min. Measurement of cigarette and e-cigarette smoke extracts and polluted water samples was used to establish proof of concept. Potentially genotoxic reactions from e- cigarette vapor similar to smoke from conventional cigarettes were demonstrated. Untreated wastewater showed a high genotoxic potential compared to negligible values for treated wastewater from a pollution control treatment plant. Reactivity of chemicals known to produce high rates of metabolite-related DNA damage were measured, and array results for environmental samples were expressed in terms of equivalent responses from these standards to assess severity of possible DNA damage. Genotoxic assessment of wastewater samples during processing also highlighted future on-site monitoring applications.
机译:开发了一种新颖的,自动化的,低成本的三维(3-D)打印微流体阵列,以检测环境样品中化学物质代谢产物对DNA的损害。电化学发光(ECL)检测平台在约10 nm深的微孔中结合了微粒体酶,DNA和发射ECL的钌金属聚合物的逐层(LbL)组装膜。液体样品被引入到阵列中,并被人的酶代谢,如果可能的话,产品会与DNA反应,并通过ECL用相机检测DNA损伤。阵列对相对DNA损伤的测量可评估样品的遗传毒性潜力。阵列在5分钟内同时分析三个样品。香烟和电子烟烟雾提取物和污水样品的测量用于建立概念证明。证明了与常规香烟烟雾相似的电子香烟蒸气潜在的遗传毒性反应。与来自污染控制处理厂的经处理废水的微不足道的值相比,未经处理的废水具有高的遗传毒性潜力。测量了已知会产生高代谢物相关DNA破坏率的化学物质的反应性,并根据这些标准的等效反应来表示环境样品的阵列结果,以评估可能的DNA破坏的严重性。加工过程中废水样品的遗传毒性评估也突出了未来的现场监测应用。

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