Mannosylation of pH-sensitive liposomes promoted cytoplasmic delivery of protein to macrophages: green fluorescent protein (GFP) performed as an endosomal escape tracer
| Autor: | Manju Kanamala, Mingtan Tang, Hao-Han Chang, Alan J. Davidson, Kaiyun Yang, Zimei Wu |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Cytoplasm
Liposome Luminescent Agents Microscopy Confocal Cell Survival Chemistry Ph sensitive liposomes Endosome Macrophages Macrophage targeting Green Fluorescent Proteins Pharmaceutical Science Endosomes General Medicine Hydrogen-Ion Concentration Green fluorescent protein carbohydrates (lipids) Mice RAW 264.7 Cells Mannosylation Liposomes Biophysics Animals Mannose |
| Zdroj: | Pharmaceutical Development and Technology. 26:1000-1009 |
| ISSN: | 1097-9867 1083-7450 |
| DOI: | 10.1080/10837450.2021.1969406 |
| Popis: | Conventional non-pH-sensitive liposomes for cytoplasmic delivery of protein suffer from poor efficiency. Here we investigated mannosylated pH-sensitive liposomes (MAN-PSL) for cytoplasmic delivery of protein to macrophages RAW 264.7 using PSL and non-pH-sensitive liposomes for comparison. We characterised the pH-dependent fluorescence of green fluorescent protein (GFP) and encapsulated it in liposomes as an intracellular trafficking tracer. GFP showed a reversed 'S'-shaped pH-fluorescence curve with a dramatic signal loss at acidic pH. GFP stored at 4 °C with light protection showed a half-life of 10 days (pH 5-8). The entrapment efficiency of GFP was dominated by the volume ratio of intraliposomal core to external medium for thin-film hydration. Mannosylation did not affect the pH-responsiveness of PSL. Confocal microscopy elucidated that mannosylation promoted the cellular uptake of PSL. For both these liposomes, the strongest, homogeneously distributed GFP fluorescence in the cytoplasm was found at 3 h, confirming efficient endosomal escape of GFP. Conversely, internalisation of non-pH-sensitive liposomes was slow (peaked at 12 h) and both Nile Red and GFP signals remained weak and punctuated in the cytosol. In conclusion, GFP performed as a probe for endosome escape of liposomal cargo. Mannosylation facilitated the internalisation of PSL without compromising their endosomal escape ability. |
| Databáze: | OpenAIRE |
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