Cytotoxicity and intracellular fate of PLGA and chitosan-coated PLGA nanoparticles in Madin–Darby bovine kidney (MDBK) and human colorectal adenocarcinoma (Colo 205) cells Mihaela Trif, 1 Paula E. Florian, 1 Anca Roseanu, 1 Magdalena Moisei, 1 Oana Craciunescu, 2 Carlos E. Astete, 3 Cristina M. Sabliov 3 1 Institute of Biochemistry of the Romanian Academy, Bucharest 060031, Romania 2 Department of Cellular Biology, National Institute R&D for Biological Sciences, Bucharest 060031, Romania 3 Biological and Agricultural Engineering Department, Louisiana State University and LSU Agricultural Center, Baton Rouge, Los Angeles 70803 Received 24 February 2015; revised 22 April 2015; accepted 6 May 2015 Published online 11 June 2015 in Wiley Online Library (wileyonlinelibrary.com). DOI: 10.1002/jbm.a.35498 Abstract: Polymeric nanoparticles (NPs) are known to facili- tate intracellular uptake of drugs to improve their efficacy, with minimum bioreactivity. The goal of this study was to assess cellular uptake and trafficking of PLGA NPs and chito- san (Chi)-covered PLGA NPs in Madin–Darby bovine kidney (MDBK) and human colorectal adenocarcinoma (Colo 205) cells. Both PLGA and Chi–PLGA NPs were not cytotoxic to the studied cells at concentrations up to 2500 lg/mL. The positive charge conferred by the chitosan deposition on the PLGA NPs improved NPs uptake by MDBK cells. In this cell line, Chi–PLGA NPs colocalized partially with early endo- somes compartment and showed a more consistent perinu- clear localization than PLGA NPs. Kinetic uptake of PLGA NPs by Colo 205 was slower than that by MDBK cells, detected only at 24 h, exceeding that of Chi–PLGA NPs. This study offers new insights on NP interaction with target cells supporting the use of NPs as novel nutraceuticals/drug delivery systems in meta- bolic disorders or cancer therapy. V C 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 103A: 3599–3611, 2015. Key Words: nanoparticle, PLGA, chitosan, epithelial cell, uptake, intracellular trafficking How to cite this article: Trif M, Florian PE, Roseanu A, Moisei M, Craciunescu O, Astete CE, Sabliov CM. 2015. Cytotoxicity and intracellular fate of PLGA and chitosan-coated PLGA nanoparticles in Madin–Darby bovine kidney (MDBK) and human colorectal adenocarcinoma (Colo 205) cells. J Biomed Mater Res Part A 2015:103A:3599–3611. INTRODUCTION Polymeric nanoparticles (NPs) have been proven to enhance the cellular uptake of entrapped bioactive compounds, to improve the drug efficacy, and to possess better stability, rel- ative to alternative delivery systems. 1–3 While many studies have consistently established successful delivery of com- pounds and therapeutic agents using biodegradable poly- meric particles, it is imperative to develop a more thorough understanding of NPs–cell interaction, in order to better pre- dict the effect of nanodelivery on the activity and availability of entrapped compounds. Polymer selection plays an important role in NPs func- tionality and bioreactivity. Poly(D,L-lactide-co-glycolide) (PLGA) is an example of common synthetic polymer used in biological applications, approved by the Food and Drug Administration (FDA) as generally recognized as safe (GRAS) for pharmaceutical applications. 4,5 Its degradation occurs mostly through hydrolysis, when it breaks down into lactic and glycolic acid. These monomers are metabolized through Krebs cycle and eliminated as harmless waste prod- ucts, carbon dioxide and water. 6,7 Various formulations of PLGA particles have been developed to promote sustained drug release, and targeting, or surface modified to increase drug absorption. 8 Chitosan is the deacetylated derivative of chitin, a natu- rally occurring biopolymer with minimal toxicity, compared to other cationic polymers such as poly-L-lysine and polye- thyleneimine. 9 Owing to its positive charge due to protona- tion at acidic pH and that promotes mucoadhesive properties, chitosan was identified as a potential agent that increased the gastrointestinal (GI) transit time of NPs and, consequently, the bioavailability of the entrapped bioactive components. Even though chitosan has not yet been approved by FDA as GRAS for food applications, it is Correspondence to: C. M. Sabliov; e-mail: csabliov@agcenter.lsu.edu Contract grant sponsor: Increasing Scientific Data on the Fate, Transport and Behavior of Engineered Nanomaterials in Selected Environmental and Biological Matrices funding program administered jointly by the Environmental Protection Agency (EPA), National Science Foundation (NSF), and US Department of Agriculture (USDA); contract grant number: EPA-G2010-STAR-N2 Food Matrices (Award No. 2010-05269) and the NSF-EPSCoR LA-SiGMA (EPS-1003897) Contract grant sponsor: Institute of Biochemistry through Romanian Academy project/2013, Structural and functional proteomics. V C 2015 WILEY PERIODICALS, INC. 3599