奈米金團簇對小鼠胚胎發育的影響

Effects of Gold-Nanoclusters on Mouse Embryo Development

洪瑋勵(Wei-Li Houng)

指導教授 ： 詹文雄

中原大學/工學院/生物科技研究所/碩士(2010年)

https://doi.org/10.6840/cycu201000943

摘要

金奈米粒子的製備是利用前置液體蝕刻成金奈米團簇。在本實驗中，我們使用金奈米粒子接枝硫酸鋅(DHLA)或聚乙烯亚胺(PEI)。因為金奈米團簇可以發出紅光，所以可採用金奈米團簇接上抗體成為免疫螢光染劑。但金奈米團簇的安全性和生物相容性還沒有完全被了解。在本實驗中，我們使用老鼠胚胎當作實驗測試平台，分析螢光金奈米團簇的安全性和細胞毒性。實驗是以囊胚期的胚胎與62.5、 125、 以及250 nM的金奈米團簇共培養24小時，以原位末端標記法、細胞增生分析和體外發育實驗等方法來探討胚胎細胞死亡、增生和發育的情形。結果，發現62.5和125 nM的金奈米團簇與胚胎共培養24小時後細胞沒有發生凋亡；進一步分析發現細胞增生與未處理的對照組並無差異，也發現胚胎的體外發育亦不受到影響。這些結果意味著金奈米團簇可成開發為胚胎發育分析的免疫螢光染劑，也有潛力開發成活體動物的螢光生物追蹤劑。

關鍵字

金奈米團簇 ； 胚胎

並列摘要

Gold nanoclusters were prepared from a one-phase reaction and etched into small nanoclusters by a gold precursor solution. In this study, gold nanoclusters were bioconjugated with DHLA or PEI. Gold nanoclusters are considered good candidates for development as luminescent probes because they have the advantages of emission of high intensity light, resistance to quenching, and good photochemical stability. These properties suggest that gold nanoclusters could be useful in various biochemical assays, especial immunofluorescence staining. However, cytotoxicity of gold nanoclusters is unclear and need further investigation. In this thesis, we examine the cytotoxic effects of gold nanoclusters, a novel luminescent material, on early postimplantation embryonic development in mouse blastocysts. Mouse blastocysts were incubated in medium with or without gold nanoclusters (62.5、125、or 250 nM) for 24 h. Cell apoptosis was analyzed by TUNEL assay and proliferation was investigated by hochest staining. Preimplantation and postimplantation development was assessed by in vitro analyses. In this thesis, we show that gold nanoclusters (250–500 nM) induces apoptotic processes, inhibits cell proliferation and impairs embryonic development in mouse blastocysts, but exerts no injury effects at treatment dosages below 250 nM. These results provide important information in terms of dosage safety and biocompatibility of gold nanoclusters to facilitate its further use as a fluorescence bio-tracer preparation.

並列關鍵字

Gold-Nanoclusters ； embryo

參考文獻

1. Aubin-Tam, M.E. and K. Hamad-Schifferli, Gold nanoparticle cytochrome c complexes: The effect of nanoparticle ligand charge on protein structure. Langmuir, 2005. 21(26): p. 12080-12084.

2. Bhattacharya, J., et al., Gold nanoparticle-based tool to study protein conformational variants: implications in hemoglobinopathy. Nanomedicine-Nanotechnology Biology and Medicine, 2007. 3(1): p. 14-19.

3. Boyer, C., et al., Anti-fouling magnetic nanoparticles for siRNA delivery. Journal of Materials Chemistry, 2010. 20(2): p. 255-265.

4. Chah, S., M.R. Hammond, and R.N. Zare, Gold nanoparticles as a colorimetric sensor for protein conformational changes. Chemistry & Biology, 2005. 12(3): p. 323-328.

5. Chan, W.H. and Shiao, N.H. Cytotoxic effect of CdSe quantum dots on mouse embryonic development. Acta Pharmacologica Sinica, 2008. 29(2): p. 259-266.