環保專案成果報告查詢系統

專案計畫編號	EPA-98-U1U1-02-104
經費年度	098
計畫經費	1,850 千元
專案開始日期	2009/04/07
專案結束日期	2009/12/31
專案主持人	張章堂
主辦單位	永續發展室(停用)
承辦人	吳婉怡
執行單位	國立宜蘭大學
專案分類	廢棄物管理
中文關鍵字	奈米微粒、細胞存活能力、細胞凋亡試驗
英文關鍵字	nano-particle,TiO2, SiO2
協同主持人	鄧燕妮、賴裕順、莊一全
共同主持人
計畫聯絡人	張章堂
計畫聯絡電話	03-9357400#
計畫聯絡信箱	ctchang@niu.edu.tw

中文摘要	機車引擎排放廢氣中含大量有機奈米微粒，而工業易排放無機奈米微粒，此些奈米微粒易透呼吸系統與皮膚進入身體內部。本研究乃藉由細胞模式，暴露於奈米微粒(含有機奈米微粒與無機奈米微粒)下，評估奈米微粒對於人類呼吸道細胞的生長影響。　　本研究是利用溶膠凝膠法合成或水熱法合成，在不同燒結時間與溫度作用下，得到不同粒徑TiO2或SiO2奈米微粒。有機奈米微粒則由機車引擎廢氣所排放，並予以分徑求得。另外，本研究亦以定量輸出霧化器及電噴霧氣膠產生器產生奈米粒徑範圍多粒徑分布( polydisperse ) 及單粒徑分布的氣懸微粒。在細胞毒性檢測部份，研究結果發現不論是有機或無機奈米顆粒，對細胞生長的影響均有抑制與促進增生現象發生，其中以TiO2 300nm(500ug/ml)對NIH/3T3細胞抑制生長最明顯可高達90%的抑制效果，進一步分析這些奈米顆粒抑制細胞生長的機制，利用annexin V來分析細胞是否有進行細胞凋亡，結果顯示所有的奈米顆粒並不會誘導細胞凋亡，而是可能直接造成細胞壞死來抑制細胞生長。　　奈米顆粒促進細胞增生方面，有機奈米微粒Oganic-10nm對A549以及NIH/3T3有明顯細胞增生的現象，TiO2_10nm與300nm粒徑則分別對HacaT與BEAS-2B細胞株具顯著的細胞增生，對細胞促進增生的情況都可高達50%，進一步利用細胞癒合測試來分析奈米顆粒是否會促使細胞進行轉型癌化，結果顯示所有奈米顆粒並不具促使細胞轉型癌化的能力。　　綜合以上結果我們發現不論是有機或無機奈米微粒都會造成細胞毒性，特別是在某些粒徑大小的奈米分子會造成特殊細胞生長特性的改變，而細胞生長改變的方式分別有抑制和促進增生的現象，但尚未發現有轉型癌化的情況。此外我們此次研究發現奈米顆粒如TiO2與SiO2有凝集的現象，此現象是否會對細胞攝入奈米微粒產生影響仍待確定。我們已設計一套良好的奈米顆粒暴露模式，希望未來能應用確保細胞株皆能正確地反映出遭受奈米微粒暴露後的機制。
中文關鍵字	奈米微粒、細胞存活能力、細胞凋亡試驗

中文摘要

機車引擎排放廢氣中含大量有機奈米微粒，而工業易排放無機奈米微粒，此些奈米微粒易透呼吸系統與皮膚進入身體內部。本研究乃藉由細胞模式，暴露於奈米微粒(含有機奈米微粒與無機奈米微粒)下，評估奈米微粒對於人類呼吸道細胞的生長影響。　　本研究是利用溶膠凝膠法合成或水熱法合成，在不同燒結時間與溫度作用下，得到不同粒徑TiO2或SiO2奈米微粒。有機奈米微粒則由機車引擎廢氣所排放，並予以分徑求得。另外，本研究亦以定量輸出霧化器及電噴霧氣膠產生器產生奈米粒徑範圍多粒徑分布( polydisperse ) 及單粒徑分布的氣懸微粒。在細胞毒性檢測部份，研究結果發現不論是有機或無機奈米顆粒，對細胞生長的影響均有抑制與促進增生現象發生，其中以TiO2 300nm(500ug/ml)對NIH/3T3細胞抑制生長最明顯可高達90%的抑制效果，進一步分析這些奈米顆粒抑制細胞生長的機制，利用annexin V來分析細胞是否有進行細胞凋亡，結果顯示所有的奈米顆粒並不會誘導細胞凋亡，而是可能直接造成細胞壞死來抑制細胞生長。　　奈米顆粒促進細胞增生方面，有機奈米微粒Oganic-10nm對A549以及NIH/3T3有明顯細胞增生的現象，TiO2_10nm與300nm粒徑則分別對HacaT與BEAS-2B細胞株具顯著的細胞增生，對細胞促進增生的情況都可高達50%，進一步利用細胞癒合測試來分析奈米顆粒是否會促使細胞進行轉型癌化，結果顯示所有奈米顆粒並不具促使細胞轉型癌化的能力。　　綜合以上結果我們發現不論是有機或無機奈米微粒都會造成細胞毒性，特別是在某些粒徑大小的奈米分子會造成特殊細胞生長特性的改變，而細胞生長改變的方式分別有抑制和促進增生的現象，但尚未發現有轉型癌化的情況。此外我們此次研究發現奈米顆粒如TiO2與SiO2有凝集的現象，此現象是否會對細胞攝入奈米微粒產生影響仍待確定。我們已設計一套良好的奈米顆粒暴露模式，希望未來能應用確保細胞株皆能正確地反映出遭受奈米微粒暴露後的機制。

中文關鍵字

奈米微粒、細胞存活能力、細胞凋亡試驗

類型	檔名	檔案大小	說明	最新版公開日期
期末報告	期末報告最終版.pdf	4MB		2010/3/1 上午 12:00:00

摘要	Motorcycle engine exhaust emissions containing large amounts of organic nanoparticles, while industrial emissions of inorganic nanoparticles easily, these nanoparticles easily enter into body by the respiratory system and skin. In this study, the human respiratory cell lines were exposed to the organic and inorganic nanoparticles to investigate the effects of cell growth. 　　The sol-gel and hydrothermal synthesis method were use to produceTiO2 or SiO2 nanoparticles by different sintering time and temperature. Organic nanoparticles from motorcycle engine exhaust emissions were subpath obtained. In addition, the study has given quantitative output of plastic atomizer and EFI fog generator produces nano-size range of multi-particle size distribution (polydisperse) and single-particle size distribution of aerosol particles. Four respiratory-related cell lines, A549, HEp2, HacaT, BEAS-2B and a non-transformation of NIH/3T3 were used for cell toxicity study. The inorganic nanoparticleTiO2, SiO2 have not serious toxicity for these cel lines. However, the results of MTT assay find that some nanoparticles have a significant effect on cell growth, such asTiO2 and SiO2 on NIH/3T3 cell lines significantly inhibited the growth, 10nm and 50nm ofTiO2 particle have a significant growth inhibition of the BEAS-2B cell lines, and while the diesel exhaust extract filtrate 13 markedly inhibited HEp2 cell growth. The nanoparticles induced cell death by necrosis pathway, not apoptosis, in the annexin V assay. In addition, we found that some nanoparticles would significantly promote cell growth, such as 10nm Organic nanoparticles for A549 and NIH/3T3 and 10nm and 300 nmTiO2 for HacaT and BEAS-2B cells. Most of nanoparticles do not have transformation activity for human respiratory cell lines. 　　Based on the above results, we found that most of the nanoparticles do not cause cell toxicity and only some specific nanoparticles can cause characteristic changes for specific cell lines, while changes including cells inhibition and proliferation phenomenon. In addition, we have design a cell exposure model to prevent aggregation for nanoparticle and make ensure that the cell lines to correctly reflect the effect of nanoparticles.
英文關鍵字	nano-particle,TiO2, SiO2

摘要

　Motorcycle engine exhaust emissions containing large amounts of organic nanoparticles, while industrial emissions of inorganic nanoparticles easily, these nanoparticles easily enter into body by the respiratory system and skin. In this study, the human respiratory cell lines were exposed to the organic and inorganic nanoparticles to investigate the effects of cell growth. 　　The sol-gel and hydrothermal synthesis method were use to produceTiO2 or SiO2 nanoparticles by different sintering time and temperature. Organic nanoparticles from motorcycle engine exhaust emissions were subpath obtained. In addition, the study has given quantitative output of plastic atomizer and EFI fog generator produces nano-size range of multi-particle size distribution (polydisperse) and single-particle size distribution of aerosol particles. Four respiratory-related cell lines, A549, HEp2, HacaT, BEAS-2B and a non-transformation of NIH/3T3 were used for cell toxicity study. The inorganic nanoparticleTiO2, SiO2 have not serious toxicity for these cel lines. However, the results of MTT assay find that some nanoparticles have a significant effect on cell growth, such asTiO2 and SiO2 on NIH/3T3 cell lines significantly inhibited the growth, 10nm and 50nm ofTiO2 particle have a significant growth inhibition of the BEAS-2B cell lines, and while the diesel exhaust extract filtrate 13 markedly inhibited HEp2 cell growth. The nanoparticles induced cell death by necrosis pathway, not apoptosis, in the annexin V assay. In addition, we found that some nanoparticles would significantly promote cell growth, such as 10nm Organic nanoparticles for A549 and NIH/3T3 and 10nm and 300 nmTiO2 for HacaT and BEAS-2B cells. Most of nanoparticles do not have transformation activity for human respiratory cell lines. 　　Based on the above results, we found that most of the nanoparticles do not cause cell toxicity and only some specific nanoparticles can cause characteristic changes for specific cell lines, while changes including cells inhibition and proliferation phenomenon. In addition, we have design a cell exposure model to prevent aggregation for nanoparticle and make ensure that the cell lines to correctly reflect the effect of nanoparticles.

英文關鍵字

nano-particle,TiO2, SiO2