Novel Green Biosynthesis of 5-Fluorouracil Chromium Nanoparticles Using Harpullia pendula Extract for Treatment of Colorectal Cancer

Abstract

Colorectal cancer (CRC) is the third highest major cause of morbidity and mortality worldwide. Hence, many strategies and approaches have been widely developed for cancer treatment. This work prepared and evaluated the antitumor activity of 5-Fluorouracil (5-Fu) loaded chromium nanoparticles (5-FuCrNPs). The green biosynthesis approach using Harpullia (H) pendula aqueous extract was used for CrNPs preparation, which was further loaded with 5-Fu. The prepared NPs were characterized for morphology using scanning and transmission electron microscopes (SEM and TEM). The results revealed the formation of uniform, mono-dispersive, and highly stable CrNPs with a mean size of 23 nm. Encapsulation of 5-Fu over CrNPs, with a higher drug loading efficiency, was successful with a mean size of 29 nm being produced. In addition, Fourier transform infrared (FTIR) and X-ray diffraction pattern (XRD) were also used for the investigation. The drug 5-Fu was adsorbed on the surface of biosynthesized CrNPs in order to overcome its clinical resistance and increase its activity against CRC cells. Box-Behnken Design (BBD) and response surface methodology (RSM) were used to characterize and optimize the formulation factors (5-Fu concentration, CrNP weight, and temperature). Furthermore, the antitumor activity of the prepared 5-FuCrNPs was tested against CRC cells (CACO-2). This in vitro antitumor study demonstrated that 5-Fu-loaded CrNPs markedly decreased the IC50 of 5-Fu and exerted more cytotoxicity at nearly all concentrations than 5-Fu alone. In conclusion, 5-FuCrNPs is a promising drug delivery system for the effective treatment of CRC.

Keywords: 5-Flourouracil; chromium nanoparticles; colorectal cancer; green biosynthesis.

Figure 1

Schematic diagram showing the details of green biosynthesis preparation method, where, (CrNPs) is Chromium nanoparticles, (5-Fu) 5-Fluorouracil, and (5-FuCrNPs) 5-Fluorouracil chromium nanoparticles.

Figure 2

(a). X-ray powder diffraction (XRD) patterns of the prepared CrNPs. (b). The Williamson-Hall (W-H) plots for CrNPs. The calculations of (β_correct) were carried out according to Psd-Voigt distribution functions.

Figure 3

Scanning electron micrographs of the prepared (a) CrNPs, (b) 5-FuCrNPs N1 and (c) N10.

Figure 4

Transmission electron microscope analysis of the prepared CrNPs with a diameter distribution histogram.

Figure 5

Transmission electron microscope analysis of the prepared 5-FuCrNPs with a diameter distribution histogram.

Figure 6

Fourier transform infrared spectra of the prepared CrNPs, 5-FuCrNPs, H. pendula extract, and 5-Fu.

Figure 7

Surface plots effects of different formulation factors on LE%.

Figure 8

Surface plots effects of different formulation factors on Release after one h.

Figure 9

Surface plots effects of different formulation factors on release after 1 h.

Figure 10

Viability of the CACO cells treated with different concentrations of 5-Fu and 5-FuCrNPs determined by the MTT assay (a). Flow cytometry analysis showing the frequency of apoptotic cells treated with 5-Fu (b), CrNPs (c), and 5-FuCrNPs (d).