Chitosan Nanoparticles-Preparation, Characterization and Their Combination with Ginkgo biloba Extract in Preliminary In Vitro Studies

Abstract

Nanoparticles (NPs), due to their size, have a key position in nanotechnology as a spectrum of solutions in medicine. NPs improve the ability of active substances to penetrate various routes: transdermal, but also digestive (active endocytosis), respiratory and injection. Chitosan, an N-deacetylated derivative of chitin, is a natural biodegradable cationic polymer with antioxidant, anti-inflammatory and antimicrobial properties. Cross-linked chitosan is an excellent matrix for the production of nanoparticles containing active substances, e.g., the Ginkgo biloba extract (GBE). Chitosan nanoparticles with the Ginkgo biloba extract (GBE) were obtained by ion gelation using TPP as a cross-linking agent. The obtained product was characterized in terms of morphology and size based on SEM and Zeta Sizer analyses as well as an effective encapsulation of GBE in nanoparticles-FTIR-ATR and UV-Vis analyses. The kinetics of release of the active substance in water and physiological saline were checked. Biological studies were carried out on normal and cancer cell lines to check the cytotoxic effect of GBE, chitosan nanoparticles and a combination of the chitosan nanoparticles with GBE. The obtained nanoparticles contained and released GBE encapsulated in research media. Pure NPs, GBE and a combination of NPs and the extract showed cytotoxicity against tumor cells, with no cytotoxicity against the physiological cell line.

Keywords: Ginkgo biloba extract; chitosan nanoparticles; cytotoxicity; drug release.

Figure 1

SEM images of Ch(GB)NPs (a) at 12,000× magnification and (b) at 120,000× magnification.

Figure 2

Dependence of the intensity on the size of nanoparticles in the tested Ch(GB)NPs sample, (a) without initial sonification and (b) with a 10 min initial sonification.

Figure 2

Dependence of the intensity on the size of nanoparticles in the tested Ch(GB)NPs sample, (a) without initial sonification and (b) with a 10 min initial sonification.

Figure 3

The rate of release of Ginkgo biloba extract from Ch(GB)NPs in water, pH 7.0 (A) and in saline, pH 5.8 (B).

Figure 4

FTIR-ATR spectra for GBE (A), pure chitosan powder (B) and Ch(GB)NPs (C).

Figure 5

Effects of GBE, ChNPs and Ch(GB)NPs on HGF-1, HeLa and PEA1 cell viability, as determined by the MTT assay after 24 h. Values represent the means ± SD as a percentage (%) of the control.