硫酸化軟骨素與幾丁聚醣錯合物對小紅莓制放及抗癌效能之探討

本研究中，我們利用天然多醣體硫酸化軟骨素與幾丁聚醣，由於帶有不同電荷，可以將抗癌藥物小紅莓 (DOX)，藉由靜電作用力的方式結合成錯合物；為了更穩定控制其釋放情形，我們將高水溶性的硫酸化軟骨素修飾上具有反應雙鍵的官能基 (Methacrylate , MA )，利用自由基反應方式，進行交聯反應，使奈米粒子結構更加緻密，以達到穩定控制釋放結果。錯合物分析上，我們觀察包覆DOX前後粒徑大小及介面電位變化，結果發現粒徑大小沒有明顯改變，而介面電位在包覆後明顯增加，說明此劑型可以成功的包覆DOX。藥物的包覆率 (E.E% )約為70-80 %之間，在藥物釋放系統上，比較有無交聯之釋放速率，結果顯示藉由交聯過程可以有效降低藥物釋放速率。最後探討包覆DOX的奈米粒子與DOX在A549、KB細胞之毒殺效能，結果顯示經由包覆處理的DOX，對毒殺癌細胞效果更好；我們更進一步觀察細胞週期變化 (cell cycle)，結果顯示在A549、KB細胞中，包覆處理的DOX可以增加細胞週期停滯在G2/M期；接著我們利用流式細胞儀觀察細胞胞噬作用 (cell uptake)，也得到相同的結果，包覆處理的DOX被細胞所吞噬的量也較多，進而呈現較強的螢光表現，其中又以加熱處理的交聯劑型的螢光位移最多。在共軛聚焦顯微鏡實際觀察細胞內噬情形，也得到一致的結論，最後我們利用毛細管電泳 (capillary electrokinetic)進行定量分析，實際計算細胞內DOX的含量。

關鍵字

幾丁聚醣；硫酸化軟骨素；小紅莓；藥物釋放

並列摘要

In this study, two natural polysaccharides, chondrotin sulfate and chitosan, bearing different charges were used to incorporate doxorubicin (DOX) though the electrolytic interactions. In order to stabilize the complexes and increase the sustain-released efficacy of DOX, methacrylated functional groups were grafted onto chondrotin sulfate which could further proceed to a crosslinking reaction via the free radical mechanism. The particle sizes did not change significantly after loading DOX into complexes but the zeta potential of the complexes increased profoundly. This situation implies that the DOX molecules have been successfully incorporated. The encapsulation efficiency of the drug was in the range of 70-80%. The release rate of DOX from the cross-linked complexes reduced significantly. The cells killed by DOX dramatically in the complex formulation compared to free DOX in both A549 and KB cancer cell lines. The consistent results were also evidenced in flow cytometric and confocal laser scanning microscopic studies, where the cellular uptake of DOX is better after formulation. The best formulation is the one with the cross-linking treatment. Finally, the quantitative DOX content inside the cells was measured by capillary electrokinetic analysis.

並列關鍵字

Chitosan ； Chondroitin sulfate ； Doxorubicin ； Drug delivery

參考文獻

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