Systematic development of and synergistic antiinflammatory activity assessment from oil in water nanosized emulsions entrapped with two different drugs

Abstract

The objectives of the current investigation are (1) to optimize the RP-HPLC conditions using newlinelinear and non-linear models for quantifying simultaneously the chosen two drugs (chloroquine newlinephosphate and flavopiridol) in combined drug solutions, dual drug loaded nanosized emulsions, newlinein vitro drug release and stability assessment, (2) to optimize a formula with the help of linear newlineand non-linear models not only for creating bi-compartmental structures in the dispersed oil newlinedroplets of emulsions but also for entrapping two different drugs, (3) to evaluate the in vitro newlinephysicochemical properties (mean particle size, zeta potential, polydispersity index, drug newlineentrapment efficiency, in vitro drug release and stability study) of dual drug-loaded nanosized newlineemulsions, (4) to perform the preclinical toxicity studies using in vitro RBC lysis and MTT newlineassay, (5) to see the uptake of emulsion droplets/particles in macrophage cell line (Raw264.7 newlinecells) for verifying the particle internalization process, (6) to explore the in vitro anti-inflammatory potentials of emulsion and solution formulations (consist of both single drug and newlinedual drug) through immunofluorescence assay using RAW264.7 cells visualized under CLSM, newline(7) to study the in vivo inflammation reducing activities of emulsion and solution formulations newline(consist of both single drug and dual drug) using complete Freund s adjuvant (CFA)-induced newlinearthritis model in Wistar rats through percentage change in right knee joint diameter values, newlineradiographical assessment (x-ray images), ELISA and RT-PCR, (8) to verify the newlinehistomorphological changes after the hematoxylin and eosin (H and E) staining of right knee newlinejoints collected from the arthritic rats following treatment with emulsion and solution newlineformulations (consist of both single drug and dual drug) and (9) to find out the anti-rheumatic newlinetherapeutic efficacies of emulsion formulations (consist of both single drug and dual drug) via newlinemetabolomic analysis of rat plasma samples by LC-MS/MS (triple quadrupole). newline newlineFollowing entrapment of chloroquin