Methotrexate-Loaded PEGylated Chitosan Nanoparticles: Synthesis, Characterization, and in Vitro and in Vivo Antitumoral Activity

Abstract

Cancer nanotherapeutics are rapidly progressing and being implemented to solve several limitations of conventional drug delivery systems. In this paper, we report a novel strategy of preparing methotrexate (MTX) nanoparticles based on chitosan (CS) and methoxypoly(ethylene glycol) (mPEG) used as nanocarriers to enhance their targeting and prolong blood circulation. MTX and mPEG-conjugated CS nanoparticles (NPs) were prepared and evaluated for their targeting efficiency and toxicity in vitro and in vivo. The MTX-mPEG-CS NP size determined by dynamic light scattering was 213 ± 2.0 nm with a narrow particle size distribution, and its loading content (LC %) and encapsulation efficiency (EE) were 44.19 ± 0.64% and 87.65 ± 0.79%, respectively. In vitro release behavior of MTX was investigated. In vivo optical imaging in mice proved that MTX was released from particles subsequently and targeted to tumor tissue, showing significantly prolonged retention and specific selectivity. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay obviously indicated that the higher inhibition efficiency of MTX-mPEG-CS NPs meant that much more MTX was transferred into the tumor cells. A significant right-shift in the flow cytometry (FCM) assay demonstrated that MTX-loaded nanoparticles were far superior to a pure drug in the inhibition of growth and proliferation of Hela cells. These results suggest that MTX-mPEG-CS NPs could be a promising targeting anticancer chemotherapeutic agent, especially for cervical carcinoma.

Inclusion of a pH-responsive amino acid-based amphiphile in methotrexate-loaded chitosan nanoparticles as a delivery strategy in cancer therapy

“…The clinical efficacy of the conventional treatments is often compromised by the acquisition of resistance in cancer cells and/or by the generation of several side effects to the patients (Banerjee et al 2002;Dong and Mumper 2010). In this context, the nanotechnology-based pharmaceutical products might provide a wide range of new tools and possibilities in cancer therapy, from earlier diagnostics to more efficient and more targeted treatments (Chen et al 2014;Das and Sahoo 2011;Li et al 2012). …”

“…Therefore, CS nanoparticles (NPs) are of great interest as drug delivery systems, including applications for cancer therapy (Chen et al 2014;Deng et al 2014;Trapani et al 2011). The ionotropic gelation technique is particularly suitable for the incorporation of pharmaceuticals, as it can be achieved in aqueous conditions (Agnihotri et al 2004;Berger et al 2004;Fan et al 2012).…”

“…Moreover, the formation process is solely based on the electrostatic interaction of oppositely charged compounds and, thus, it is not necessary any chemical modification. CS-based NPs containing MTX were already reported in the literature, but most of them with drawback of using a cross-linked agent (Chen et al 2014;Jia et al 2014;Wu et al 2009). Likewise, there is considerable progress already achieved regarding the mechanisms underlying drug release from the nanostructures, being most of the current release methods based on reactions that commonly occur in response to environmental factors, 4 i.e.…”

Self-assembled polysaccharide nanostructures for controlled-release applications

“…Methotrexate (MTX) is a folate inhibitor used for the treatment of certain cancers and autoimmune diseases [53].…”

“…For these reasons, nanoparticle DDS have been considered as a means of improving solubility, cellular uptake, and tumor targeting through the EPR effect. Chen et al [53] prepared chitosan-based nanoparticles to improve delivery of MTX. The crosslinked chitosan nanoparticles were synthesized using a combination of ionic crosslinking (with sodium triphosphate) and covalent crosslinking (with glutaraldehyde).…”