Development and characterization of lipid nanoparticles to assess anti-inflammatory effect of therapeutic drugs

Abstract

Inflammation is one of the defensive responses of the body that involves blood vessels, immune cells, molecular mediator like chemokine, cytokine, vasoactive amines, eicosanoid (prostaglandins, leukotrines), and products of proteolytic cascades. The inflammation decreases initial cause of injury of cell, and clears necrotic cells and damaged tissues aimed at initiating tissue repair mechanism. Chronic inflammation leads to many complex diseases such as hay fever, periodontitis, atherosclerosis, rheumatoid arthritis, and cancer. Present study is an orchestrated attempt to formulate and characterized lipid based delivery vehicle, liposomes. This work was designed to achieving sustained and controlled delivery of methotrexate and aceclofeanc through liposomes to address various systemic inflammatory diseases. We used MDA-MB-468 breast cancer cells to mimic inflammation by LPS treatment for in-vitro estimation of inflammation. LPS has been reportedly known to induce inflammation characterized by pro-inflammatory cytokines such as TNF-α, IL-6 and IL-1β. Although, anti-inflammatory drugs including methotrexate (MTX) and aceclofenac (Ace) are widely used to subside inflammation, short half-life of drugs 0.7-5.8hr(MTX)and 4 hrs (Ace), lack of target specificity, bio-distribution in kidney, liver and other organs and toxicity rendered to healthy cells refrains their use. Therefore, nanocarriers especially liposomes have been introduced due to lower toxicity, bio-compatibility and capability to deliver both hydrophilic and lipophilic drug. Present study was aimed at formulating (by thin film hydration method) and characterizing liposomes. The drug loaded size was within the range of 199.3±0.3 to 285.6±0.2 nm and loading efficiencies are estimated to be 10.73% and 10.7% for MTX and Ace respectively. Further, MTT assay were carried out to calculate IC50. The levels of pro-inflammatory cytokines was estimated inform the mimicked breast cancers cells before and after therapeutic treatment with free drug and drug loaded onto liposomes. We have seen the sustained release of drug when delivered through liposome. Collectively, our results advocates for the sustained and controlled release of MTX and Ace via liposomes and open new vistas to address inflammatory diseases.