FRET-based analysis on the fate of liposome and cyclodextrin@ liposome nanocomposites from ocular surface to the posterior segment of the eye
Abstract
Investigating the structural integrity of nanocarriers in vivo is vital for exploring the fate of nanocarriers from ocular surface to the posterior segment of the eye. Most of the published studies adopted the structural integrity ratio of nanocarriers to determine the fate of them, which lacked scientific support. In this study, two methods were used to explore the factors which affected the structural integrity of liposomes. A new method with the standard curve of FRET fluorescence intensity and carbocyanine 7 (Cy7) content was drawn for the first time. Secondly, we used the traditional method of drawing the standard curve of FRET fluorescence efficiency and structural integrity ratios. The results showed that liposomes with particle size about 120?nm, positively charged, polyethyleneglycol5000 (PEG5000) and glycine sarcosine (GS) modified had the highest Cy7 content in rabbit tissues. When the dosage of Cy7 was 25?μg, at 60?min, the content of Cy7 in intact liposomes and the structural integrity ratio of liposomes in sclera was 210.5?±?14.9?ng and 19.8?±?5.3?%, respectively. Compared with the structural integrity ratio, the Cy7 content in the intact carrier could better estimate the fate of nanocarriers in vivo scientifically. On this basis, the fate of dual-carrier nanocomposites and the inner cyclodextrin complexes in vivo was investigated. The intact cyclodextrin complexes could reach choroid-retina with the protection of outer liposomes. The structural integrity ratios of liposomes were also studied after crossing human conjunctival epithelial cells (HConEpiC) monolayer, but in vitro cellular experiments could not simulate the real situation in vivo. Finally, the tissue distribution of nanocomposites was studied in rabbit eyes. The concentration of dexamethasone (Dex) in choroid-retina was 158?±?23?ng/g after 3?h, which exhibited better drug delivery ability compared with our previous study. Overall, the present study provides a new scientific method to estimate the structural integrity in vivo, which is beneficial for the rational design of drug delivery systems with more structural integrity in vivo and higher drug delivery efficiency.