Carbohydrate Polymers 94 (2013) 309–316 Contents lists available at SciVerse ScienceDirect Carbohydrate Polymers jo u rn al hom epa ge: www.elsevier.com/locate/carbpol Self-assembled nanoparticles based on amphiphilic chitosan derivative and hyaluronic acid for gene delivery Ya Liu, Ming Kong, Xiao Jie Cheng, Qian Qian Wang, Li Ming Jiang, Xi Guang Chen ∗ College of Marine Life Science, Ocean University of China, Qingdao, 266003, PR China a r t i c l e i n f o Article history: Received 13 September 2012 Received in revised form 18 December 2012 Accepted 21 December 2012 Available online 5 January 2013 Keywords: OCMCS HA DNA NPs Gene vector a b s t r a c t The present work described nanoparticles (NPs) made of oleoyl-carboxymethy-chitosan (OCMCS)/hyaluronic acid (HA) using coacervation process as novel potential carriers for gene delivery. An N/P ratio of 5 and OCMCS/HA weight ratio of 4 were the optimal conditions leading to the smallest (164.94 nm), positive charged (+14.2 mV) and monodispersed NPs. OCMCS-HA/DNA (OHD) NPs showed higher in vitro DNA release rates and increased cellular uptake by Caco-2 cells due to the HA involved in NPs. The MTT survival assay indicated no significant cytotoxicity. The transfection efficiency of OHD NPs was 5-fold higher than OCMCS/DNA (OD) NPs; however, it decreased significantly in the presence of excess free HA. The results indicated that OHD NPs internalized in Caco-2 cells were mediated by the hyaluronan receptor CD44. The data obtained in the present research gave evidence of the potential of OHD NPs for the targeting and further transfer of genes to the epithelial cells. © 2012 Elsevier Ltd. All rights reserved. 1. Introduction Gene delivery is a promising strategy as the encoded protein can be expressed in the host in its natural form (without dena- turation or modification), and caused prolonged expression of the protein (Mao, Sun, & Kissel, 2010) The gastrointestinal (GI) system is an excellent target for noninvasive localized drug delivery due to its large surface area and its accessibility. For this reason, the GI tract has been considered as an attractive target for gene therapy interventions (Nandedkar, 2009). The success in gene therapeutic strategies depends on an efficient system for the delivery of nucleic acid into the target cells (Vadolas, Williamson, & Ioannou, 2002). In recent years, non-viral vectors and especially those resulting from the use of nanotechnologies have received increasing attention for achieving the delivery of genetic material to the GI tract (Luten, van Nostrum, De Smedt, & Hennink, 2008). Cationic polymers have been shown as promising carriers among the non-viral gene delivery systems. Many cationic poly- mers, such as chitosan, polylysine, polyethyleneimine, dendrimers, poly (a-(4-aminobutyl)-l-glycolic acid) as well as cationic lipo- somes have been investigated for gene delivery (Mohammadi et al., 2011). Chitosan has been investigated as nonviral vector for gene ∗ Corresponding author at: College of Marine Life Science, Ocean University of China, 5# Yushan Road, Qingdao 266003, China. Tel.: +86 0532 82032586; fax: +86 0532 82032586. E-mail addresses: xgchen@ouc.edu.cn, qqsn160@163.com (X.G. Chen). delivery because of its ability to condense gene into nanoparticles that are appropriate to be endocytosed by cells, and subsequently be released from endosomes and enter nucleus (Muzzarelli, 2010a). Furthermore, chitosan is an ideal candidate for oral DNA delivery due to its good biocompatibility and high positive charge density conferring it mucoadhesive properties (Lai, Wang, & Hanes, 2009; Muzzarelli, 2010b). However, chitosan shows two major disadvantages: one is poor solubility because the amino groups on chitosan are only partially protonized at physiological pH 7.4. The other is low transfection efficiency (Gao et al., 2008). In our previous research, oleoyl-carboxymethy-chitosan (OCMCS) has been synthesized and proposed as one of water-soluble chitosan derivatives over a wide pH range (Liu, Cheng, et al., 2012; Liu, Zang, et al., 2012). The transfection efficiency of chitosan vectors can be improved by combining chitosan with cationic or anionic biopolymers, such as polyethyleneimine (Zhao et al., 2009) or arginine (Gao et al., 2008), prior to the addition of DNA. The choice of biopolymer greatly influences the specificity, stability, and size of the assem- bled nanoparticles (Duceppe & Tabrizian, 2009). Hyaluronic acid (HA) is another biocompatible anionic biopolymer naturally found in humans and is used for a great number of clinical applica- tions (Muzzarelli, Greco, Busilacchi, Sollazzo, & Gigante, 2012). This biopolymer has some advantageous properties, including its ability to bind various cellular receptors such as CD44 (Aruffo, Stamenkovic, Melnick, Underhill, & Seed, 1990), which is expressed in normal human epithelium cells, chondrocytes and cancerous cells (Marhaba & Zoller, 2004). The targeting of the gene carriers 0144-8617/$ – see front matter © 2012 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.carbpol.2012.12.058