Comparative study of the in vitro and in vivo characteristics of cationic and neutral liposomes

Abstract

Wei Zhao, Song Zhuang, Xian-Rong QiDepartment of Pharmaceutics, School of Pharmaceutical Sciences, Peking University, Beijing, People's Republic of ChinaAbstract: Neutral liposomes (NLP) exhibit preferential localization in solid tumors based on the enhanced permeation and retention (EPR) effect. Cationic liposomes (CLP) have a propensity for localizing in newly formed tumor vessels and they have a potentially enhanced antitumor effect. However, an increased amount of cationic lipids in liposomes also induces aggregation through electrostatic interactions between the liposomes and the anionic species in the circulation, which results in a reduced EPR effect. Consequently, it is important to investigate the characteristics of liposomes with different surface potentials in vitro to achieve an optimal intratumoral distribution and antitumor effect in vivo. In this study, the authors evaluated the characteristics of doxorubicin (DOX)-loaded NLPs, CLPs, polyethylene glycol (PEG)-modified NLPs (NLP-PEG), and PEGylated CLPs (CLP-PEG) (ie, encapsulation efficacy, zeta potential, size, membrane fluidity, aggregation in serum, and uptake of liposomes into rat aortic endothelial cells (RAECs)) to further understand their influences on the pharmacokinetics, biodistribution, and antitumor therapy in vivo. The results showed that increased amounts of cationic lipids resulted in severe liposome aggregation in the presence of serum, yet it did not alter the membrane fluidity to a large extent. The uptake of liposomes into RAECs, visualized by confocal fluorescence microscopy, confirmed the rapid uptake of CLP by the endothelial cells compared with NLP. However, the pharmacokinetics, biodistribution and anticancer efficacies of these liposomes in vivo revealed that the CLP with highly positive surface potentials exhibited reduced circulation times and poor distribution in tumors. The NLP-PEG exhibited the highest anticancer efficacy; CLP-PEG, the second highest; and CLP with the most positive surface potential, the lowest. These phenomena were mostly due to the rapid aggregation in serum and subsequent accumulation in the lungs upon the intravenous injection of the CLP. Caution should be exercised when chemotherapeutic drugs are loaded into CLP for tumor therapy.Keywords: cationic liposomes, PEGylation, electrostatic interaction, aggregation, anticancer efficacy, biodistribution