Department of Pharmaceutical Sciences, Center for Pharmaceutical Biotechnology & Nanomedicine, Northeastern University, 140 Fenway Street, Boston, MA 02115, USA.
Nanomedicine (Lond). 2013 Jul;8(7):1055-65. doi: 10.2217/nnm.12.138. Epub 2012 Dec 2.
We hypothesized that liposomes modified with lysosomotropic octadecyl-rhodamine B (Rh) and loaded with therapeutic glucocerebroside velaglucerase alfa (VPRIV™) will improve lysosomal delivery of the enzyme into Gaucher's cells.
MATERIALS & METHODS: Confocal microscopy and flow cytometry were used to evaluate the ability of Rh-modified liposomes loaded with VPRIV to improve the lysosomal targeting in monocyte-derived macrophages and Gaucher's fibroblasts.
Confocal microscopy demonstrated that Rh-modified liposomes localized primarily in the lysosomes. As confirmed by flow cytometry using specific substrate 5-(pentafluorobenzoylamino)fluorescein diglucoside, intralysosomal accumulation of VPRIV in the cells treated with Rh-modified liposomes was significantly increased (up to 68%) relative to the cells treated with plain liposomes or free VPRIV.
Rh-modified lysosomotropic liposomes can improve lysosomal accumulation of liposomal enzymes both in nonphagocytic Gaucher's fibroblasts and phagocytic monocyte-derived macrophages.
我们假设用溶酶体靶向性十八烷基罗丹明 B(Rh)修饰并负载治疗性葡糖脑苷脂酶维拉苷酶阿尔法(VPRIV™)的脂质体将改善酶进入戈谢细胞的溶酶体递送。
共聚焦显微镜和流式细胞术用于评估负载 VPRIV 的 Rh 修饰脂质体改善单核细胞衍生巨噬细胞和戈谢氏纤维母细胞中溶酶体靶向的能力。
共聚焦显微镜显示 Rh 修饰的脂质体主要定位于溶酶体中。如使用特异性底物 5-(五氟苯甲酰氨基)荧光素二葡萄糖苷通过流式细胞术所证实的,与用普通脂质体或游离 VPRIV 处理的细胞相比,用 Rh 修饰的脂质体处理的细胞中 VPRIV 的溶酶体内蓄积显著增加(高达 68%)。
Rh 修饰的溶酶体靶向脂质体可以改善非吞噬性戈谢氏纤维母细胞和吞噬性单核细胞衍生巨噬细胞中脂质体酶的溶酶体蓄积。