Contents lists available at ScienceDirect Journal of Microbiological Methods journal homepage: www.elsevier.com/locate/jmicmeth Flurbiprofen-loaded ethanolic liposome particles for biomedical applications Sarvesh Paliwal a , Amita Tilak a , Jaiprakash Sharma b , Vivek Dave a, ⁎ , Swapnil Sharma a , Kanika Verma a , Kajal Tak a , Kakarla Raghava Reddy c , Veera Sadhu d, ⁎ a Department of Pharmacy, Banasthali Vidyapith, Rajasthan, India b Department of Pharmacy, SMS Medical College, Rajasthan, India c School of Chemical and Biomolecular Engineering, The University of Sydney, Sydney, NSW 2006, Australia d School of Physical Sciences, Banasthali Vidyapith, Banasthali, Rajasthan, India ARTICLE INFO Keywords: Flurbiprofen Ethanolic liposome particles Transdermal drug delivery Analgesic Anti-inflammatory ABSTRACT Present study deals with the preparation, characterization and in-vivo evaluation of flurbiprofen loaded etha- nolic liposome which provides predetermined and controlled release of drug through a transdermal drug de- livery system. Ethanolic liposomes were prepared by using flurbiprofen, phospholipon 90-G, and ethanol in varied concentration ratio. The prepared ethanolic liposomes were optimized and characterized for particle size, zeta potential, polydispersive index and % entrapment efficiency. FTIR study was performed to analyze the interaction between drug and excipient. To study the thermal behavior of the formulation DSC and TGA were carried out. The surface morphology of ethanolic liposome was performed with the help of SEM, TEM, and AFM. In-vitro drug permeation study of the optimized formulation was carried out using the albino rat skin model and peripheral nociceptive activity was evaluated by writhing assay. In addition, formulations were also inspected for stability study for three months at a different temperature. The optimized formulation EF5 exhibited a particle size of 167.2 ± 3.7 nm with a zeta potential of -51.6 ± 0.2 mV and PDI of 0.209. The optimized formulation showed an ideal surface morphology with a maximum % entrapment efficiency i.e. 93.51 ± 2.1. In- vitro permeation study shows a release of 70.23% in 24 h and transdermal flux was found as 238.2 μg/cm 2 /h. Writhing assay demonstrate that the optimized formulation decreases the number of writhes and thus shows the peripheral analgesic activity. In stability study, optimized formulation showed maximum stability at 4 °C. These results suggest that transdermal system mediated application of flurbiprofen loaded ethanolic liposome can be considered as an effective way to afford consistent and predictable release of flurbiprofen which could provide beneficial effects in the management of various inflammatory diseases. 1. Introduction Transdermal drug delivery system offers some key advantages over other routes of drug administration. In particular, when a transdermal patch is applied to the skin it delivers a drug across skin through passive diffusion at a predetermined and controlled rate to attain an optimal therapeutic concentration in a subject (Dave et al., 2010). Phenylalk- anoic acid derivative flurbiprofen is a non-steroidal anti-inflammatory drug which acts via cyclo‑oxygenase inhibition which in turn inhibits prostaglandin synthesis. Prostaglandin has been considered as a prin- cipal marker of inflammation, pain, swelling, fever. Flurbiprofen is widely indicated in the treatment of various pathological conditions like acute and chronic rheumatoid arthritis, osteoarthritis and ankly- losing spondylitis. Ethanolic liposome was first developed and reported by Touitou and her colleagues in 2000 (Touitou et al., 2000). An ethanolic liposome is characterized as a lipid bilayer phospholipid vesicular system primarily composed of phospholipid and excessive ethanol. Bearing high ethanol concentration, they can effectively penetrate across stratum corneum and deeper layers of skin and possess higher transdermal flux as com- pared to conventional liposome formulation which gets remain con- fined to upper layer of skin i.e. stratum corneum (Bragagni et al., 2012). Presence of ethanol in the ethanolic liposome interacts with polar head group region of membrane lipids and alters the membrane fluidity and permeability. However, enhanced permeability of ethanolic liposome is also attributed to a collective mechanism between a high concentration of ethanol, phospholipid vesicles, and skin lipids. Ethanolic liposomes have been successfully tailored for enhanced delivery of various ther- apeutic agents such as ketoprofen, aceclofenac, Testosterone, Canna- bidiol, Buspirone hydrochloride, Erythromycin, ibuprofen, Benzocaine, https://doi.org/10.1016/j.mimet.2019.04.001 Received 25 February 2019; Received in revised form 1 April 2019; Accepted 2 April 2019 ⁎ Corresponding authors. E-mail addresses: vivekdave1984@gmail.com (V. Dave), veera.sadhu@gmail.com (V. Sadhu). Journal of Microbiological Methods 161 (2019) 18–27 Available online 03 April 2019 0167-7012/ © 2019 Elsevier B.V. All rights reserved. T