Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/53556
標題: 配合即時監測環境樣品奈米微粒細胞毒性篩選技術
作者: 闕斌如
關鍵字: 生物技術, 生物科學類, 環保工程
nano particle
環境樣品
cytotoxicity
奈米微粒
細胞毒性
應用研究
摘要: Nanomaterials and nanoparticles have received considerable attention for the past decade due to their unique properties and diverse applications in technology and life sciences. Despite the widespread use of nanomaterial products, relatively few studies have been undertaken to determine the biological effects of nanoparticle exposure. Accumulating evidences have suggested that nanoparticles exhibit cytotoxicity and genotoxicity in both in vivo and in vitro studies. Given the increasing use of nanoparticles in a range of products, it has become crucial to develop a fundamental understanding of the biological effect of nanoparticle exposure. Currently, nanoparticles toxicity testing is based on in vitro studies using cell culture system; however, many problems are found to be associated with the testing system. The purpose of this project is to establish a fast screening platform to evaluate the cytotoxicity of nanoparticles that may expose human by inhalation, dermal absorption, and ingestion. This innovative screening platform is expected to replace traditional in vitro tests with more accuracy, higher sensitivity, and faster screening. Furthermore, we propose to assess the cytotoxicity of nanoparticles in the environmental air samples which are collected by an efficient single-stager wet electrostatic precipitator that has shown promising results to reduce the typical problems associated with dry nanoparticles powder commonly used for in vitro cytotoxicity analyses. In our preliminary results, we show that there is an association between concentrations of nanosilver and inhibition of cell growth analyzing by the xCELLigence system. Since we have not obtained enough cytotoxicity data evaluating by traditional MTT assays, therefore, it is difficult to make a comparison between these testing methods at this point. On the other hand, we are working on the characterization of nanoparticles collected by wet nanoparticle sampler system (WPS). Furthermore, the cytotoxic effect of nanoparticles collected from WPS will be examined by xCELLigence system.
全球奈米科技正在如火如荼的發展,此種新興科技雖然在各個領域上,開拓新的希望、應用價值與新興產品,然而這些具有奈米結構的物質,其本身的化學性質與物理性質,會隨著粒徑大小的不同而有所變化,對人體的毒性危害或影響也可能有所改變,因此對於安全衛生環保亦將可能產生重大的衝擊。雖然目前已有一些關於奈米微粒毒性研究的報導,但因為各種不同的材質,製備法及毒性測試條件的不同,造成許多不一致或互相矛盾的結論出現。而目前普遍被認為的因素可影響奈米微粒的細胞毒性有1) 奈米微粒濃度, 2) 奈米微粒粒徑大小, 3) 奈米微粒結構, 4)奈米微粒表面電荷, 5) 奈米微粒表面積以及 6) 暴露於奈米微粒環境所使用的培養液組成;再加上7) 所使用的不同的體外細胞實驗系統,及不同的生物活性分析方法或檢測方式,以及所使用的檢測方法與奈米微粒可能互有干擾等等的因素,更大幅提高實驗的複雜度及其所衍生出來的結果無法歸納成可信的定論。利用體外細胞株的實驗模式已成為最重要的檢測化學物質毒性的方法,與動物實驗模式比較的話,體外細胞株實驗提供簡單、快速、及花費經濟的優點;然而,體外細胞株實驗畢竟缺乏動物實驗系統或人體的複雜性及完整性,再加上許多細胞株的代謝能力的特性並沒有完整的被解釋清楚,造成體外細胞株的實驗模式仍有它受限之處及缺點。在探討奈米微粒的毒性及健康風險時,由於奈米微粒的物理化學性質常限制了體外細胞株毒性的檢測方法,因此發展出一套新型的具有高再現性及準確性的毒性檢測方法,就成為目前的研究重點。傳統的檢測方法大多仰賴染劑或指示劑的運用,但這些染劑或指示劑經常會與受到奈米微粒的干擾,因此新的檢測方法建立,首要為去除染劑或指示劑的使用。本研究團隊經過文獻回顧後發現,國際上目前一般仍使用傳統體外細胞毒性檢測方法,測試奈米微粒的毒性,但此種方法有其缺失及不足之處。本計劃旨在尋找其他的替代方法,以期得到更準確的實驗結果。在針對今年度的計劃建議之工作內容,我們認為可以利用細胞連續及即時生長分析儀,建立更穩定、快速及可測試大量奈米微粒樣品的篩選平台。細胞連續及即時生長分析儀的原理為,以電子感測器偵測細胞因貼附在培養皿所產生的微量電阻,細胞電阻則換算成並以Cell Index (CI) 表示,而細胞電阻或Cell Index則與細胞數目、生長狀況、細胞功能及貼附性成正比,與電腦連線的分析儀可連續性並長時間的即時觀察細胞生長與貼附性的微小變化。在目前初步的執行成果中,我們1. 建立細胞連續及即時生長分析儀的篩選平台2. 進行以傳統細胞毒性測試法分析奈米銀及奈米氧化鋅3. 進行以細胞連續及即時生長分析儀分析奈米銀與奈米氧化鋅4. 進行以流式細胞儀檢測細胞內吞噬入的奈米銀及奈米氧化鋅濃度同時,在初步的測試結果中,我們發現以細胞連續及即時生長分析儀分析法,奈米銀配置的溶液對細胞的生長抑制效果,隨著濃度增加而增加。但在奈米氧化鋅的系統下,生長抑制效果的濃度曲線則較不理想。我們將持續進行傳統毒性測試MTT方法,累積足夠的數據後,與細胞連續及即時生長分析儀分析法之結果做比較。
URI: http://hdl.handle.net/11455/53556
其他識別: EPA-99-U1U1-02-104
文章連結: http://grbsearch.stpi.narl.org.tw/GRB/result.jsp?id=2167801&plan_no=EPA-99-U1U1-02-104&plan_year=99&projkey=PG9910-0513&target=plan&highStr=*&check=0&pnchDesc=%E9%85%8D%E5%90%88%E5%8D%B3%E6%99%82%E7%9B%A3%E6%B8%AC%E7%92%B0%E5%A2%83%E6%A8%A3%E5%93%81%E5%A5%88%E7%B1%B3%E5%BE%AE%E7%B2%92%E7%B4%B0%E8%83%9E%E6%AF%92%E6%80%A7%E7%AF%A9%E9%81%B8%E6%8A%80%E8%A1%93
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