探討表面修飾磁性奈米粒子於斑馬魚動物模型之毒性驗證

Toxicity effect of uncoated and coated magnetic nanoparticles (Fe304) in vivo study

馬 禾 川(Nemi Malhotra)

指導教授 ： 葛宗融

共同指導教授 ： 蕭崇德(Chung Der Hsiao)

中原大學/工學院/生物醫學工程研究所/博士(2021年)

https://doi.org/10.6840/cycu202100259

摘要

磁性奈米顆粒（MNP）作為新型多功能的新興星星材料，正在醫學、電子、廢水處理、化妝品和能源存儲設備的不同領域中得到廣泛研究。一些MNP配方已可用於商業用途並在市場上販售。但是，MNPs的潛在生物毒性仍未得到廣泛研究。不同濃度MNP製劑對模式生物和細胞的毒性研究仍有待更多研究。在本研究中，我利用低濃度(1 ppm）與高濃度（10 ppm）的未包覆奈米Fe3O4（MNPs）和碳包覆Fe3O4（C-MNPs）進行斑馬魚的神經行為變化評價。MNPs和C-MNPs的物理特徵利用穿透式電子顯微鏡（TEM）、超導量子干涉儀（SQUID）、磁力計和振動樣品磁力計（VSM）、熱重分析（TGA）、X射線衍射（XRD）、拉曼光譜和傅立葉變換紅外光譜（FTIR）進行鑑定。與控制組相較，實驗證明經由MNPs暴露後的斑馬魚在積極性和天敵躲避測試沒有改變。在高濃度的MNPs暴露測試中觀察到緊密的群游行為。我們還注意到在MNPs處理組中，新環境水槽試驗中魚的探索行為大幅度減少，同種社會互動試驗中的魚群互動顯著減少。通過使用抗原特異性ELISA試驗，我們發現經由MNPs暴露可以顯著提高皮質醇，乙酰膽鹼（ACh）和過氧化氫酶（CAT）的水平。並降低血清素，乙酰膽鹼酯酶（AChE）和多巴胺水平。相較之下，C-MNPs比MNPs相對毒性較小，因為在成年斑馬魚的行為和生化測試中，低濃度和高濃度的C-MNPs均不會引起明顯的行為與生化指標改變。我們的數據充分表明經由碳表面包覆改質的MNPs可以大幅增加其對斑馬魚的生物安全性。我們的研究成果可為醫療保健行業的奈米顆粒的設計和劑量規範提供參考

關鍵字

斑馬魚，磁性奈米顆粒，神經行為分析，神經傳遞物質

並列摘要

Magnetic nanoparticles (MNPs) are widely being investigated as new multifunctional promising agents in different fields of the theranostics in medical sciences, electronics, waste water treatment, cosmetics and energy storage devices. Some MNPs formulations are already available in the market for commercial use. However, MNPs neurobehavioral toxicity is still not being explored extensively. Research is still in progress to analyse the toxicity effect of different concentrations and formulations of MNPs on model organisms and cell lines. In this study we attempted to determine the neurobehavioral changes in zebrafishes at low (1 ppm) or high (10 ppm) concentration of uncoated nano-Fe3O4 (MNPs) and carbon coated Fe3O4 (C-MNPs). In house synthesized uncoated MNPs and C-MNPs were characterized by transmission electron microscopy (TEM), superconducting quantum interference device (SQUID) magnetometer and vibrating sample magnetometer (VSM), thermo-gravimetric analysis (TGA), X-ray diffraction (XRD), Raman spectroscopy and Fourier transform infrared (FTIR) spectroscopy respectively. We demonstrated the behavior of zebrafish in aggression and predator avoidance were unaltered after MNPs exposure. Tight shoal groups were observed during shoaling test at high concentration of MNPs exposure. We also noted a drastic reduction in fish exploratory behaviour in novel tank test and a significant reduction in conspecific social interaction test in the MNPs treated groups. By using antigen-specific ELISA test, we found MNPs exposure can significantly elevate cortisol, acetylcholine (ACh) and catalase (CAT) levels; and reduce serotonin, acetylcholine esterase (AChE) and dopamine levels. Whereas C-MNPs were observed to be less toxic in comparison to uncoated MNPs since neither low nor high concentration of C-MNPs elicit toxicity response in performed behavioral and biochemical tests in adult zebrafish. Our data demonstrates potential toxicity of uncoated MNPs and safety of C-MNPs on adult zebrafish that have not been reported earlier. Our study can be helpful in designing and dosage specification of nanoparticles for the health-care industry.

並列關鍵字

Zebrafish, Magnetic Nanoparticles, Neurobehavioral analysis, toxicity, Neurotransmitters

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