Silymarin-loaded Eudragit 1 RS100 nanoparticles improved the ability of silymarin to resolve hepatic ﬁbrosis in bile duct ligated rats N. Younis a, *, Mohamed A. Shaheen b , Marwa H Abdallah c,d a Department of Biochemistry, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt b Department of Histology and Cell Biology, Faculty of Medicine, Zagazig University, Zagazig 44519, Egypt c Department of Pharmaceutics and Industrial pharmacy, Faculty of Pharmacy, Zagazig University, Zagazig 44519, Egypt d Department of Pharmaceutics, Faculty of Pharmacy, Hail University, Saudi Arabia A R T I C L E I N F O Article history: Received 1 March 2016 Received in revised form 26 March 2016 Accepted 28 March 2016 Keywords: Silymarin Nanopreciptation Eudragit 1 RS100 Fibrosis Bile duct ligation A B S T R A C T Some nano-formulations of silymarin (SM), a drug commonly used for liver diseases, were developed to overcome its poor solubility and poor bioavailability; antiﬁbrotic effect of these formulations has not been tested yet. In this study we aimed to formulate and evaluate silymarin-loaded Eudragit 1 RS100 nanoparticles (SMnps) and to test the capability of SMnps to reverse an established ﬁbrosis model. SMnps were prepared by solvent evaporation and nano-precipitation techniques. The inﬂuence of drug: polymer ratio, concentration of surfactant in the aqueous phase on particle size, drug entrapment efﬁciency (EE) % and in vitro drug releases were investigated. For in vivo evaluation, bile duct ligated (BDL)-rats were treated with either SM or SMnps every other day (125 mg/kg) orally for 3 weeks started 3 weeks after BDL. Liver function tests, oxidative stress and ﬁbrosis/ﬁbrogenesis process were evaluated using biochemical and histopathological techniques. The formulation No (F4) of SMnps showed an average particle size of 632.28  12.15 nm, a drug EE% of 89.47  1.65% and released the drug in a prolonged manner over 24 h. The prepared SMnps were nearly spherical with smooth surfaces. In BDL- rats, treatments with either SM or SMnps corrected liver function and oxidative stress. Only SMnps was able to reverse the induced ﬁbrosis; SMnps signiﬁcantly decreased serum tumor necrosis factor- a (TNF- a), serum transforming growth factor- b1 (TGF-b1), hepatic hydroxyproline and downregulated the hepatic expression of tissue inhibitor metalloproteinase-1 (TIMP-1) and cytokeratin-19 (CK-19), whilst increased hepatic hepatocytes growth factor (HGF) and upregulated the hepatic expression of matrix metalloproteinase-2 (MMP-2) and increased MMP-2/TIMP-1 ratio at mRNA level. Livers of rats treated with SMnps showed very little collagen in ECM and restored hepatic architecture as compared to either BDL rats or rats treated with SM. Conclusion: Formulation of silymarin as nanoparticles improved its ability to resolve cholestasis-induced liver ﬁbrosis by restoring hepatic regenerative capabilities. Therefore, formulation of SMnps may represent a step forward in the ﬁeld of anti-ﬁbrotic drug development. ã 2016 Elsevier Masson SAS. All rights reserved. 1. Introduction Several approaches have been reported to improve the in vivo performance of poorly soluble drugs when given orally depending on the reduction of drug particle size to increase the dissolution rate and the oral bioavailability of these drugs [1]. Among these approaches, nanotechnology has emerged as a powerful and promising tool. The use of nanoparticulated systems offers many advantages in drug delivery, mainly focusing on improved safety and efﬁcacy of the drugs, providing targeted delivery of drugs, prolonging drug or gene effect in target tissue and improving the stability of drug against chemical/enzymatic degradation [2]. Eudragit 1 RS100, the co-polymer of poly (ethylacrylate, methyl- methacrylate and chlorotrimethyl-ammoniumethyl methacrylate) containing quaternary ammonium group, is commonly used for the formulation of controlled and sustained release dosage forms [3]. It is insoluble in physiological pH and capable of swelling, which represents a good material for drug dispersion. Silymarin, an extract of the milk thistle (Silybum marianum), is a mixture of ﬂavonoids and polyphenols; Silibinin is its major bioactive constituent. Silymarin is a well-known hepatoprotective drug which has been shown to have antioxidant, anti- * Corresponding author. E-mail address: nahlayounis2003@yahoo.com (N. Younis). http://dx.doi.org/10.1016/j.biopha.2016.03.042 0753-3322/ ã 2016 Elsevier Masson SAS. All rights reserved. Biomedicine & Pharmacotherapy 81 (2016) 93–103 Available online at ScienceDirect www.sciencedirect.com