S. Joshi, A. Daryanani, R. Sahu, V.A. Dennis, S.R. Singh
Alabama State University,
United States
Keywords: Microfluidics, Liposomes, Staphylococcus Aureus
Summary:
Microfluidics is a novel one-step method to prepare liposomes that are reproducible, and high throughput compared to the traditional thin-film hydration method commonly used to produce liposomes. Using microfluidics, it is possible to increase the percent drug loading and achieve uniform particle size distribution at the same time. On the other hand, the staphylococcus aureus can infect many parts of the human body such as the upper respiratory tract, and skin. Currently available treatments for skin infections mostly fall in the antibiotic class of drugs; however, along with liposomes, there are other drug delivery systems such as gold and silver nanoparticles that are also being tested by researcher around the globe for the inhibition of staphylococcus aureus. In this study, garcinol (GC) an antimicrobial agent was encapsulated into the liposomes using a microfluidic device and the formulation is being tested for its efficiency in stopping the staphylococcus aureus spread when in contact with human epithelial cells. All the formulations were prepared using DSPC: Cholesterol 10:4 w/w. The liposomal formulation was tested for particle characteristics using dynamic light scattering (DLS) and percent drug encapsulation using UV-Visible spectrophotometry. MTT and wound healing assay (WHA) were performed to evaluate the toxicity to the human keratinocytes, fibroblasts, and human epithelial type-2 (HEp-2) cells. Inhibition of Staphylococcus aureus was analyzed using the Kirby-Bauer method. It was observed that the size of the liposomes was around 50 nm (with polydispersity index not more than 0.2) and not significantly affected (p>0.05) by the encapsulation of the GC (Figure 1-Additional Information). It was also observed that the liposomal encapsulation using microfluidics was 43.11 % ± 1.9. The liposome without and with GC ( 0.1, 0.2, 0.6, and 1 mM) was found non-toxic to all the three types of epithelial cells; where the cell viability was more than 80 %, and that was significantly better (p< 0.05) than GC alone (>0.2 mM), where the observed toxicity of the GC was more than 50 % (cell viability < 50 %). The WHA has confirmed also supports the results obtained by the MTT assay. Finally, the assay by Kirby-Bauer method confirmed that GC can inhibit staphylococcus aureus and liposomal formulation can be used to control the release of the GC as well as reduce its toxicity (Figure 2-Additional Information). Overall, microfluidics is the convenient, faster, and efficient method of drug encapsulation into liposomes. Uniform liposomes of size well below 100 nm can be achieved by this method. Liposomes can encapsulate GC as well as lowers its toxicity without losing the potency of the GC. Finally, GC alone and GC encapsulated into the liposomes is found better than an antibiotic (control).