以細胞株直接暴露系統應用於評估燃燒產物細胞毒性

Using Cell Direct Exposure System in Evaluating Toxicity of Combustion Product

本研究使用蕭等人改良的Air Liquid Interface, ALI（氣、液介面）暴露系統搭配燃燒系統進 行燃燒產物的細胞毒理測試，次評估ALI暴露系統應用在燃燒產物細胞毒理評估之可行性。 本研究將線香放置燃燒系統中使用高壓電源供應器對暴露系統施加電壓，讓微粒直接沉積 Transwell的細胞上，進一步使用掃瞄式電移動度粒徑分析儀監測微粒的濃度及系統的穩定度。 燃燒系統會產生三組濃度，讓細胞同時暴露混合污染物，再加裝濾網過濾微粒，讓細胞單純暴 露氣態污染物，進而比較其毒性差異。暴露完之後以trypan blue染色法及MTT assay來計算細胞 的存活率，並量測細胞釋放Reactive Oxygen Species, ROS的產量評估燃燒產物與細胞氧化性壓 力的關係。 研究結果顯示產生源的粒徑分布範圍為100~400nm，且以單峰呈現。數目中位數粒徑 介於163nm~195nm之間。本系統的沉積率在施加電壓後使微粒整體的沉積率提升5%~15%。 細胞微粒的暴露劑量與Trypan blue染色法所呈現細胞的存活率的減少率呈現顯著的線性關係 (R2=0.9501)與ROS釋放量的增加率亦呈現顯著的線性關係(R2=0.9071)。暴露氣態污染物的細胞 存活率皆高於暴露混合污染物細胞，反應出暴露混合物的毒性高過於暴露氣態污染物。 不同的暴露劑量對於Trypan blue染色法所呈現細胞的存活率的減少率及ROS釋放量的增加 率都有顯著的相關性，顯示本系統可有效使微粒暴露不同劑量燃燒產物，並可利用Trypan blue 及ROS產量評估出混合物及氣態污染物對細胞的毒性危害。但本研究在細胞上的整體沉積率後 續仍需以電子顯微鏡確認之。

One study use a modified ALI exposure system developed by Hsiao et al with combustion products from the combustion systems for cell toxicity testing. We evaluated the feasibility of the ALI exposure system using for air mixtures from combustion. The incense was placed in burning system and high voltage power supply was applied on the exposure system. To increase particles deposition on Transwell. Scanning Mobility Particle Sizer was used to measure the particles levels in the exposure system. There levels of combustion products were introduced to cell exposure chamber. Filtration system was then used to remove the particles from the system, to create a gas phase exposure scenario for ALI. Trypan blue staining and MTT assay were used to measure the cell viability, and ROS production was used to evaluate the oxidative stress from combustion products. One results shown the range of particle diameter were 100 to 400 nm with unimodal distribution. The number median diameter were between 163nm to 195nm three levels of generation source. The deposition rate of the exposure system shows the increased 5% to 15% with applying voltage. Cell viability reduction rate shown a significant decrease with increasing exposing doses in a linear relationship (R2 = 0.9501). The increase of ROS release also shown a significant linear relationship with exposing doses (R2 = 0.9071). Linear dose dependent effects also shown on gas phase exposure. However, the toxicity were lower than mixtures of combustion. One study use significantly, decrease linear dose-dependent effects deposition rate and exposure time to estimate the exposure dose for cells both could be observed on reduction rate of cell viability and the increase of ROS for mixture of combustion and only gas phase. One system can effectively use to different combustion products, showing the overall deposition rate on ALI cells should further examine by electron microscopy in the near future.