第一代與第二代樹狀高分子對小鼠囊胚之細胞毒性分析

樹枝狀高分子（Dendrimers）在生物醫學應用上越來越多,因此對它的生物特性之瞭解是相當重要的。陽離子的樹狀高分子如PAMAM會造成細胞溶血及具有細胞毒性，而且它的毒性隨著代數越大而表面基團數目越多而增加。因此低代數的樹枝狀高分子應用於當藥物傳輸的載體上來說是具有很高的發展性與價值性。研究也證實將樹枝狀高分子表面接枝上poly (etylene glycol) (PEG)可有效降低樹枝狀高分子的毒性而提高其生物相容性。為了確定樹枝狀高分子在生物醫學上使用的安全性，本研究探討與評估了第二代的PAMAM (G2)與PAMAM-PEG (G2) 樹枝狀高分子以及第一代的PAMAM (G1) 和Phosphorus (G1) 樹枝狀高分子對於老鼠囊胚期胚胎的細胞毒性。收集囊胚期的ICR老鼠胚胎並與各種濃度的PAMAM樹狀高分子(50 、 100、 200 、300 、或 400 µg/ml)共培養，透過一連串的偵測，包括TUNEL細胞凋亡分析、Hoechst染色以及囊胚期胚胎體外發育分析來探討樹枝狀高分子的細胞毒性。實驗的結果發現表面修飾PEG的第二代PAMAM樹枝狀高分子和表面是磷酸基（Phosphorus）的第一代PAMAM樹枝狀高分子比未修飾的PAMAM樹枝狀高分子或表面是氨基的PAMAM樹枝狀高分子對造成囊胚細胞凋亡與影響胚胎細胞的增生之傷害皆較低。在胚胎體外分析方面，也發現囊胚與表面修飾PEG的第二代PAMAM樹枝狀高分子共培養組也比與未修飾的PAMAM樹枝狀高分子共培養組有較好的發育情形。

關鍵字

phosphorus ； PAMAM ； PEG ；囊胚； dendrimer

並列摘要

Biological properties of dendrimers are highly important because the increased interest in using them for biomedical applications. Cationic dendrimers, like PAMAM, are generally haemolytic and cytotoxic. Their toxicity is generation-dependent and increases with the number of surface groups, suggesting that low generation dendrimers may be promising agents for targeted drug delivery. Conjugation with poly (ethylene glycol) (PEG) chains is also considered as a method of reducing the toxicity and increasing the biocompatibility of dendrimers. In this study to ensure its safety in biomedical usage, the low generation of PAMAM (G2) and PAMAM-PEG (G2) dendrimer, also PAMAM (G1) and Phosphorus (G1) dendrimer are evaluated for their cytotoxicity on mouse blastocysts. Embryos of ICR mouse in blastocyst stage are collected and exposed to various concentrations of all kinds of dendrimer (50 µg/ml, 100 µg/ml, 200 µg/ml, 300 µg/ml, and 400 µg/ml) and evaluated via TUNEL assay, Hoechst staining, and blastocyst outgrowth experiment. Blastocysts that were treated with PAMAM-PEG (G2) dendrimer and phosphorus (G1) dendrimer showing less apoptotic lesions and more cell proliferation number compared with the PAMAM (G2) and PAMAM (G1) respectively. The PEGylated dendrimer also showed better outgrowth (≥80% grade III outgrowth) compared with the unmodified one

並列關鍵字

blastocyst ； dendrimer ； phosphorus ； PEG ； PAMAM

參考文獻

Albert, B., Johnson, A., Lewis, J., Raff, M., Roberts, K., & Walter, P. (2002). Molecular biology of the cell, 4th edition. New York: Garland Science.

Anggelia, M. R. (2012). Gisenoside-Rg1 exposure induces developmental injury and intrinsic apoptotic pathway in mouse embryos (master’s theses). Chungli: Chung Yuan Christian University.

Armant, D. R., Kaplan, H. A., & Lennarz, W. J. (2004). Fibronectin and laminin promote in vitro attachment and outgrowth of mouse blastocysts. Developmental Biology, 116(2), 519-523.

Caminade, A. M., & Majoral, J. P. (2013). Positively charged phosphorus dendrimers. An overview of their properties. New Journal of Chemistry, 37(11), 3358-3373.

Carayannopoulos, M. O., Chi, M. M. Y., Cui, Y., Pingsterhaus, J. M., McKnight, R. A., Mueckler, M., . . . Moley, K. H. (2000). GLUT8 is a glucose transporter responsible for insulin-stimulated glucose uptake in the blastocyst. Proceedings of the National Academy of Sciences, 97(13), 7313-7318.

國際替代計量

第一代與第二代樹狀高分子對小鼠囊胚之細胞毒性分析

全文下載

主題瀏覽