- 學位論文
以PLGA/Liposomes微粒包覆去氧核醣核酸水解酶之研究
Research of DNase encapsulated by PLGA/Liposomes microspheres
摘要
傳統的藥物傳遞系統大多以藥錠、膠囊、針劑等方式呈現，而近年來因為生物可分解性材料聚乳酸-聚甘醇酸(poly(DL-lactide-co-glycolide)，PLGA)被廣泛研究，發展出PLGA微粒包覆藥物之新型藥物傳遞系統。本研究先利用微脂粒包覆去氧核醣核酸水解酶(Deoxyribonuclease，DNase)，並探討出包覆率較佳之配方，再利用PLGA包覆製備完成之微脂粒，之後將活性最佳之微粒與巨噬細胞共培養，觀察微粒被巨噬細胞吞噬的比例。 結果顯示各種微脂粒配方中，以添加氯化鈣的微脂粒配方有最佳的包覆率45.13% (以蛋白量為基準)及43.79% (以活性為基準)，而在微粒釋放研究中發現，以PLGA包覆添加10 mM氯化鈣、蝦紅素及海藻糖之微脂粒配方有最佳的累積活性，經過五天的釋放後，每毫克的微粒活性可釋放出有32.59 U，該配方之微粒與巨噬細胞共培養的結果顯示，在投入微粒的初期，巨噬細胞的吞噬現象並不明顯，投入微粒共培養12小時後，約有22.84%的微粒被吞噬，而當巨噬細胞吞噬一定量的微粒後，即會停止吞噬現象，吞噬微粒後的巨噬細胞，其存活率約為90.41%，該結果顯示，微粒被吞噬後對於巨噬細胞的存活影響並不大，不會造成細胞大量死亡。
並列摘要
The traditional drug delivery systems include pills, capsules, or injection. In this study, A novel drug delivery system utilizing poly (DL-lactide-co- glycolide)(PLGA) encapsulation technology was developed. Deoxyribonuclease (DNase) was encapsulated in egg yolk phosphatidylcholine (EYPC) liposomes with stabilizers, such as charged lipids, astaxanthin and trehalose. The result showed that DNase stabilized in CaCl2 solution and then encapsulated with EYPC liposomes showed the highest encapsulation efficiency(45.13%) and enzyme activity(43.79%). DNase-liposomes was then encapsulated in PLGA microspheres. PLGA microspheres containing DNase, which stabilized with CaCl2, and encapsulated with astaxanthin, and trehalose stabilized liposomes, showed the highest DNase activity after release. The accumulated DNase activity can reach 32.56 unit/mg microspheres. The microspheres were then cultured with macrophages. Macrophages phagocytosis was not significant in the primary stage. After 12 hr culture, about 22.84 % of microspheres were phagocyted by macrophages. We found that macrophages phagocytosis stopped when 43% microspheres were engulfed. The viability of macrophages was 90.41%, after 48 hr culture, suggesting the microspheres will not lead to death of macrophages.