Bacterial count (CFU/ml) on different days of incubation of a 7 day biofilm of <i>K</i>. <i>pnuemoniae</i> B5055 in a microtiter plate following treatment with plain liposomes (empty lipoosmes as control), antibiotic (amikacin at 40 μg/ml), non-entrapped phages (MOI = 1) and liposome entrapped phages (MOI = 1).

<p>All values represent the mean ± SEM, calculated from two independent experiments, each performed in duplicate on different occasions.</p

Encapsulation of Bacteriophage in Liposome Accentuates Its Entry in to Macrophage and Shields It from Neutralizing Antibodies

<div><p>Phage therapy has been a centre of attraction for biomedical scientists to treat infections caused by drug resistant strains. However, ability of phage to act only on extracellular bacteria and probability of interference by anti-phage antibodies <i>in vivo</i> is considered as a important limitation of bacteriophage therapy. To overcome these hurdles, liposome were used as delivery vehicle for phage in this study. Anti-phage antibodies were raised in mice and pooled serum was evaluated for its ability to neutralize free and liposome entrapped phage. Further, ability of phage and liposome-entrapped phage to enter mouse peritoneal macrophages and kill intracellular <i>Klebsiella pneumoniae</i> was compared. Also, an attempt to compare the efficacy of free phage and liposome entrapped phage, alone or in conjunction with amikacin in eradicating mature biofilm was made. The entrapment of phage in liposome provided 100% protection to phage from neutralizing antibody. On the contrary un-entrapped phage got neutralized within 3 h of its interaction with antibody. Compared to the inability of free phage to enter macrophages, the liposome were able to deliver entrapped phage inside macrophages and cause 94.6% killing of intracellular <i>K</i>. <i>pneumoniae</i>. Liposome entrapped phage showed synergistic activity along with amikacin to eradicate mature biofilm of <i>K</i>. <i>pneumoniae</i>. Our study reinforces the growing interest in using phage therapy as a means of targeting multidrug resistant bacterial infections as liposome entrapment of phage makes them highly effective <i>in vitro</i> as well as <i>in vivo</i> by overcoming the majority of the hurdles related to clinical use of phage.</p></div

Comparison of effect of neutralizing antibodies on phage and liposome entrapped phage.

<p>Comparison of effect of neutralizing antibodies on phage and liposome entrapped phage.</p

Particle size distribution and PDI of liposomal formulation of phage.

<p>Particle size distribution and PDI of liposomal formulation of phage.</p

Comparison of percent killing of intracellular bacteria when treated with phage and liposome entrapped phage.

<p>Comparison of percent killing of intracellular bacteria when treated with phage and liposome entrapped phage.</p

Bacterial count (CFU/ml) on different days of incubation of a 7 day biofilm of <i>K</i>. <i>pnuemoniae</i> B5055 in a microtiter plate following treatment with plain liposomes (empty lipoosmes as control), antibiotic (amikacin at 40 μg/ml), non-entrapped phages (MOI = 1) and liposome entrapped phages (MOI = 1).

<p>All values represent the mean ± SEM, calculated from two independent experiments, each performed in duplicate on different occasions.</p

Percent bacterial uptake and killing by mouse peritoneal macrophages at different time intervals.

<p>All values represent the mean ± SEM, calculated from two independent experiments, each performed in duplicate on different occasions. SEM represent the 95% confidence interval. Confidence interval is calculated as SEM * 3.18. Raw data is available in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0153777#pone.0153777.t002" target="_blank">Table 2</a>.</p

Biodistribution of liposome entrapped phage and unentrapped phage in spleen of BALB/c mice (n = 3).

<p>All values represent the mean ± SEM, calculated from two independent experiments, each performed in duplicate on different occasions</p

Bacterial count (CFU/ml) on different days of incubation of a 7 day biofilm of <i>K</i>.<i>pneumoniae</i> following treatment with phage + antibiotic (P+A) and liposome entrapped bacteriophage + antibiotic (LP+A) in a microtiter plate.

<p>All values represent the mean ± SEM, calculated from two independent experiments, each performed in duplicate on different occasions. A- Amikacin (40 μg/ml), P- Phage, LP- Liposome entrapped phage.</p

Microemulsions mediated effective delivery of methotrexate hydrogel: <i>more than a tour de force</i> in psoriasis therapeutics

<p>Methotrexate (MTX), a well known drug for the treatment of cancer and rheumatoid arthritis, has gained prominence in the treatment of psoriasis over the period of years. However, the present mode of systemic administration through oral or parenteral route has always proposition, full of compromises. The toxicity of drug to the vital organs and physiological environment is the major concern. Also, its poor skin penetration is one major problem. Hence novel system based on lipid carriers has been considered here to overcome the barriers. Microemulsions (MEs) were prepared using pseudo-ternary phase diagram (PTPD) and they were characterized for various parameters such as size, shape (cryo-SEM), PDI, zeta potential, etc. The chosen MEs system (optimized) was then incorporated into secondary vehicles and characterized for rheological behavior, texture profile analysis, <i>in vitro</i> release, <i>ex vivo</i> permeation and drug distribution into different layers of skin. The developed formulations were further evaluated in <i>ex vivo</i> and <i>in vivo</i> such as cell line study, imiquimod-induced psoriatic model, allergic contact dermatitis, rat tail model (% orthokeratosis) and safety test (Draize test). The MEs based MTX gel has shown its potential in locating the drug at the desired domain of stratum corneum, epidermal and dermal layers of skin and reducing systemic absorption. Our results are suggestive of MEs potential as a novel carrier for topical delivery of MTX in topical therapeutic and safety approaches. In conclusion, developed MEs-based hydrogel has shown promising results in achieving effective delivery of MTX.</p