Arginylglycylaspartic Acid-Surface-Functionalized Doxorubicin-Loaded Lipid-Core Nanocapsules as a Strategy to Target Alpha(V) Beta(3) Integrin Expressed on Tumor Cells

Abstract

Doxorubicin (Dox) clinical use is limited by dose-related cardiomyopathy, becoming more prevalent with increasing cumulative doses. Previously, we developed Dox-loaded lipid-core nanocapsules (Dox-LNC) and, in this study, we hypothesized that self-assembling and interfacial reactions could be used to obtain arginylglycylaspartic acid (RGD)-surface-functionalized-Dox-LNC, which could target tumoral cells overexpressing αvβ3 integrin. Human breast adenocarcinoma cell line (MCF-7) and human glioblastoma astrocytoma (U87MG) expressing different levels of αvβ3 integrin were studied. RGD-functionalized Dox-LNC were prepared with Dox at 100 and 500 mg·mL−1 (RGD-MCMN (Dox100) and RGD-MCMN (Dox500)). Blank formulation (RGD-MCMN) had z-average diameter of 162 ± 6 nm, polydispersity index of 0.11 ± 0.04, zeta potential of +13.2 ± 1.9 mV and (6.2 ± 1.1) × 1011 particles mL−1, while RGD-MCMN (Dox100) and RGD-MCMN (Dox500) showed respectively 146 ± 20 and 215 ± 25 nm, 0.10 ± 0.01 and 0.09 ± 0.03, +13.8 ± 2.3 and +16.4 ± 1.5 mV and (6.9 ± 0.6) × 1011 and (6.1 ± 1.0) × 1011 particles mL−1. RGD complexation was 7.73 × 104 molecules per nanocapsule and Dox loading were 1.51 × 104 and 7.64 × 104 molecules per nanocapsule, respectively. RGD-functionalized nanocapsules had an improved uptake capacity by U87MG cells. Pareto chart showed that the cell viability was mainly affected by the Dox concentration and the period of treatment in both MCF-7 and U87MG. The influence of RGD-functionalization on cell viability was a determinant factor exclusively to U87MG.

Nanoformulation Shows Cytotoxicity against Glioblastoma Cell Lines and Antiangiogenic Activity in Chicken Chorioallantoic Membrane

“…All formulations were prepared by interfacial deposition of preformed polymer followed by surface coating and functionalization, as previously reported [29,35,36]. To obtain doxorubicin-loaded-poly(ε-caprolactone)-alpha bisabolol-LNC (AB-DOX-LNC-L), 0.1 g of a mixture of PLC 80 and PCL 10 (9:1, w/w) was dissolved in acetone (25 mL) (organic phase I, OP-I).…”

“…It is important to mention that the alpha-bisabolol amount used in this study was selected according to a previous study that determined the ideal proportion between sorbitan monostearate, oil, and polymer (1:4.1:2.6, w/w/w) [19], while doxorubicin and RGD concentrations were based on the antitumor effect previously observed for RGDfunctionalized doxorubicin-nanocapsules [29].…”

“…Surface functionalized LNCs can be produced by coating with lecithin, chitosan, and polysorbate 80, followed by interfacial reactions with metal ions (Fe 2+ , Zn 2+ , or Au 3+ ), which allows the complexation with different ligands: anti-LDL(−)-single-chain variable antibody fragment for inhibiting atheroma progression [25,26], laronidase for treating mucopolysaccharidosis type I [27], methotrexate showing enhanced and selective antiproliferative effects against human breast cancer cells [20], bevacizumab demonstrating in vitro cytotoxicity against the C6 cell line and in vivo antiangiogenic activity [28], and arginylglycylaspartic acid (RGD) as a ligand in doxorubicin-loaded nanocapsules [29]. In this last study, Pareto charts indicated that breast cancer (MCF-7) and glioma (U87MG) cell viabilities were primarily affected by the drug concentration and treatment duration and that RGD-surface functionalization is an important factor in reducing U87MG cell viability, compared with the viability in MCF-7 cells, due to the overexpression of αvβ3 integrin in glioma cells.…”

Oral Treatment of Spontaneously Hypertensive Rats with Captopril-Surface Functionalized Furosemide-Loaded Multi-Wall Lipid-Core Nanocapsules

“…A few years ago, our research group proposed a new process to functionalize the surface of chitosan-coated lipid-core nanocapsules using a ligand-metal ion-chitosan-lecithin complex [27][28][29][30][31][32][33]. This method is based on a rapid one-pot synthesis without needing any purification step, overcoming the drawbacks of other methodologies.…”

“…When the drug or the pro-drug were acting as ligands at the nanocapsule surface, a higher uptake by MCF-7 cells and a higher antiproliferative effect have been observed compared to the control or to the formulation functionalized with a non-active model of ligand (phenylalanine). In parallel, we developed doxorubicin-loaded nanocapsules having RGD as ligand on the surface to target tumoral cells overexpressing αvβ3 integrin [33]. An improved uptake of the nanocapsules by U87MG cells and a significant decrease in cell viability compared to control have been observed.…”