Synthesis and Characterization of Chitosan-Based Nanodelivery Systems to Enhance the Anticancer Effect of Sorafenib Drug in Hepatocellular Carcinoma and Colorectal Adenocarcinoma Cells

Abstract

The formation of two nanodelivery systems, Sorafenib (SF)-loaded chitosan (SF-CS) and their folate-coated (SF-CS-FA) nanoparticles (NPs), were developed to enhance SF drug delivery on human Hepatocellular Carcinoma (HepG2) and Colorectal Adenocarcinoma (HT29) cell lines. The ionic gelation method was adopted to synthesize the NPs. The characterizations were performed by DLS, FESEM, TEM, XRD, TGA, FTIR, and UV-visible spectroscopy. It was found that 83.7 ± 2.4% and 87.9 ± 1.1% of encapsulation efficiency; 18.2 ± 1.3% and 19.9 ± 1.4% of loading content; 76.3 ± 13.7 nm and 81.6 ± 12.9 nm of hydrodynamic size; 60-80 nm and 70-100 nm of TEM; and FESEM sizes of near-spherical shape were observed, respectively, for SF-CS and SF-CS-FA nanoparticles. The SF showed excellent release from the nanoparticles under pH 4.8 PBS solution, indicating a good delivery system for tumor cells. The cytotoxicity study revealed their better anticancer action towards HepG2 and HT29 cell lines compared to the free sorafenib. Moreover, both NPs systems showed negligible toxicity to normal Human Dermal Fibroblast adult cells (HDFa). This is towards an enhanced anticancer drug delivery system with sustained-release properties for better cancer management.

Keywords: HDFa; HT29; HepG2; Sorafenib; cancer; cell lines; chitosan-nanoparticles; drug-delivery; folic acid; therapeutic.

Figure 1

Schematic diagram of the synthesis steps of Sorafenib-loaded chitosan nanoparticles.

Figure 2

Schematic diagram of the synthesis steps of folic acid-conjugated chitosan Sorafenib nanoparticles.

Figure 3

Formation of chitosan-folic acid conjugation through the amine group of chitosan and OH group of FA.

Figure 4

The effect of TPP (mg/mL) on particle size (nm) and PDI index of SF-CS NPs.

Figure 5

Particles size distribution by the intensity and cumulation of (a) sorafenib-loaded chitosan nanoparticles (SF-CS) and (b) folate-conjugated sorafenib-loaded chitosan nanoparticles (SF-CS-FA).

Figure 6

XRD patterns of folic acid-coated chitosan-loaded sorafenib nanoparticles (a), folic acid (b), Sorafenib-loaded chitosan nanoparticles (c), chitosan nanoparticles (d), and the drug Sorafenib (e).

Figure 7

FESEM images and EDX spectrum of (a) CS-NPs, (b) SF-CS nanoparticles, (c) SF-CS-FA nanoparticles.

Figure 8

HRTEM micrographs of (a) Sorafenib-loaded chitosan nanoparticles (SF-CS) and (b) folate-conjugated, Sorafenib-loaded chitosan nanoparticles (SF-CS-FA).

Figure 9

FTIR spectrum of SF (a), CS-NPs (b), SF-CS NPs (c), FA (d), and SF-CS-FA NPs (e).

Figure 10

TGA/DTG thermograms of (a) CS-NPs, (b) SF, (c) SF-CS, (d) FA, and (e) SF-CS-FA.

Figure 10

TGA/DTG thermograms of (a) CS-NPs, (b) SF, (c) SF-CS, (d) FA, and (e) SF-CS-FA.

Figure 10

TGA/DTG thermograms of (a) CS-NPs, (b) SF, (c) SF-CS, (d) FA, and (e) SF-CS-FA.

Figure 11

Release profiles of SF from its (a) SF-CS and (b) SF-CS-FA nanoparticles at pH 7.4 and 4.8 buffer solutions.

Figure 12

The data fitting of SF release from its SF-CS nanoparticles at pH 4.8 (a), pH 7.4 (b) and its SF-CS-FA nanoparticles at pH 4.8 (c), pH 7.4 (d) using the pseudo-second-order kinetics models.

Figure 13

Cytotoxicity assay of chitosan, pristine sorafenib, CS-sorafenib, and CS-sorafenib-folic acid nanoparticles against normal HDFa dermal fibroblast cells at 72 h. Values are expressed as mean ± SD of triplicates. The significant differences were determined using the one-way ANOVA followed by Duncan’s Multiple Range Test.

Figure 14

Cytotoxicity assay of CS, SF, SF-CS, and SF-CS-FA nanoparticles against HepG2 cells at 72 h of incubation. Values are expressed as mean ± SD of triplicates. The significant differences (p < 0.05) * were determined using the one-way ANOVA followed by Duncan’s Multiple Range Test.

Figure 15

Cytotoxicity assay of CS, SF, SF-CS, and SF-CS-FA nanoparticles against HT29 cells at 72 h of incubation. Values are expressed as mean ± SD of triplicates. The significant differences (p < 0.05) * were determined among untreated HT29 using the one-way ANOVA followed by Duncan’s Multiple Range Test.