## 4 Discussion
The presence of GSH in liposomes loaded with EFA did not influence the physicochemical properties when compared with liposomes loaded with EFA alone (Table 2). Earlier, encapsulation of cytochrome C, a hydrophilic protein was, along with lipophilic vitamin E, developed to obtain more efficient drug encapsulations and examine their release using the same lipidmixture [47], whereas using a single lipid such as phospholipid 90H, lipophilic resveratrol was encapsulated along with a hydrophilic vitamin C to investigate the ameliorative antioxidant effect [48]. HIV-1 infection harbors the virus in latent reservoirs such as macrophages, one of the key targets of HIV-1 infected patients. Macrophages, commonly referred to as virus reservoirs sites, are not killed by the HIV infection, thus aiding the virus to reproduce and supply the body with new virus [49]. It is well known that macrophages play a crucial role in the host defense against HIV-1 infection, as they produce multiple intracellular HIV restriction factors [50]. Studies have proved that the intracellular concentrations of antiretovirals were significantly lower in macrophages than in T lymphocytes [51]. In this purview, THP-1 Mo/Mac cells were used to examine the uptake, cytotoxicity and ROS-induced fluorescence of eduirenz when delivered both as pure form and via liposomes with and without GSH. Firstly, intracellular lysate concentrations revealed a significantly higher amount of liposomal EFA taken up by the cells in comparison to pure form. The choice of co-delivering GSH was made on the basis of the evidence that supplementing GSH will result in a restoration of intracellular rGSH and a corresponding restoration of macrophage functions [2, 6, 52]. Earlier studies also suggest that GSH-liposomes were the most protective since they increased cell viability from 28% (blank liposomes) to 67% when encapsulated within liposomes [53]. Thus, as a supplement to ARV therapy, GSH-replenishing agents may attenuate the toxic effects of HAART. Previously, it has been demonstrated that the administration of high doses of GSH was able to reduce the progression of the disease, providing additional effects to azidothymidine (AZT) therapy using a murine model [54, 55]. As depicted in Figures 1 and 2, the presence of GSH (depicted as EFA + GSH) enhanced the uptake of EFA when the cells were treated either in pure form (2.46 fold) or in liposomal form (1.68 fold). Liposome delivery to such pathogen reservoirs has received successful attention in the past few years, which is also thoroughly reviewed and discussed [19, 26]. In our study, EFA liposomes were negatively charged with particle size less than 150 nm, which could have led to a higher uptake than the pure form. While greater uptake of small liposomes (<100 nm) by mononuclear phagocytic system MPS cells has been reported in the literature [56], various other studies have shown liposome uptake by MPS cells to be improved with increased size [19]. Second, MTS assay data further revealed that encapsulation of EFA into liposomes could be an alternative approach to conventional EFA forms to reduce cytotoxicity; results demonstrated 15.2 fold increase in the viability, thereby improving the proliferation of non-infected THP-1 cells. This increase in the cell viability appears to be due to encapsulation into lipid vesicles based on the assumption that liposome membranes either fuse with cell membranes or are endocytosed by the cells and release their content intracellularly [57]. This mechanism could also depend on the particular liposome formulation, thereby suggesting that negative charge improved endocytosis. Additionally, the antioxidant defense system, due to the presence of GSH within the liposomal vesicle, could have led to a reduction of oxidative stress (Figure 3).