Polycladia myrica-based delivery of selenium nanoparticles in combination with radiotherapy induces potent in vitro antiviral and in vivo anticancer activities against Ehrlich ascites tumor

Abstract

Background: Over the last few decades, nanotechnology has entered daily life through various applications, therefore, there has been a trend toward developing new approaches to green-mediated nanotechnology that encourage nanomaterial formation through biological methods such as plants or microorganisms. Algae have gained increasing attention from nanotechnology scientists and have paved the way for the emergence of "algae nanotechnology" as a promising field. Methods: Via using the aqueous extract of the brown alga Polycladia myrica, selenium nanoparticles were synthesized and characterized by using seven instruments: SEM, TEM, UV spectra, Zeta potential, EDX, X-ray diffraction, and FTIR. P. myrica selenium nanoparticles (PoSeNPs) were then examined for their antiviral activity against HSV-1 (Herpes simplex I) and anticancer against human colon cancer cell line (HCT-116) in vitro and in vivo alone and in combination with laser therapy of power 2 mW against Ehrlich carcinoma (EAC). Results: PoSeNPs ranging between 17.48 nm and 23.01 nm in size, and EDX revealed the selenium mass and its atoms as 0.46% ± 0.07% and 0.08% ± 0.01% respectively. Their anticancer potentiality in vitro was with maximum inhibitions of 80.57% and 73% and IC₅₀ = 14.86 μg/mL and 50 mg/mL against HCT-116 and EAC cell lines respectively, while their in vivo alone and in combination with laser therapy of power 2 mW showed a potent therapy effect against Ehrlich ascites carcinoma (EAC). Conclusion: This study concluded that PoSeNPs do not have a toxic effect; they exhibit high effectiveness as a photothermal agent for cancer therapy, with promising applications in future biomedical fields. The combined therapy showed a significant decrease in tumor volume, massive tumor cell necrosis, shrinking, and disappearance. It also showed improvement in liver TEM, histology, kidney function: urea and creatinine, and liver enzymes: ALT, and AST.

Keywords: EAC; HCT-116; Polycladia myrica; antiviral; in vitro; in vivo; laser therapy; selenium nanoparticles.

FIGURE 1

PoSeNPs produced some background noise in XRD pattern.

FIGURE 2

TEM imaging of PoSeNPS, showing different particle sizes and morphology (A) at scale bar (100 nm) and (B) at scale bar (200 nm).

FIGURE 3

SEM imaging of PoSeNPS (500 nm) with different particle sizes.

FIGURE 4

EDX analysis of PoSeNPS.

FIGURE 5

Field of EDX analysis of PoSeNPs.

FIGURE 6

Cytotoxicity of PoSeNPS toward HCT-116 cell line. (A) is the control HCT-116 without PoSeNPS, and (B) is HCT-116 at a 50 μg/mL concentration.

FIGURE 7

Effect of PoSeNPs on cell viability of EAC cells determined by MTT assay. Cell viability of EAC cells treated with different concentrations (ranging from 1 to 50 mg/mL) of SeNPs. Data are expressed as the mean ± SD, *** indicate very high significant difference as compared with the control treatment (p < 0.001).

FIGURE 8

Effect of different treatments showing change in the relative tumor volume.

FIGURE 9

Survival rate of the mice after different treatments.

FIGURE 10

Histological analysis of tumor injury stained with hematoxylin and eosin (H&E) (Scale bare:50 µm). (A) Tumor without treatment showing inflammatory cells (blue circle) (B) Tumor irradiated with laser showing inflammatory cells (blue circle) (C) injected tumor with PoSeNPs showing inflammatory cells (blue circle) (D) injected tumor with PoSeNPs then irradiated by laser beam showing necrosis cells (blue arrow).

FIGURE 11

Photomicrographs of sections in liver stained by H&E (A) Liver of negative control group showing normal hepatic architecture (red arrow) (scale bare:50 µm). (B) Liver of mice bearing Ehrlich carcinoma (positive control group) showing birding portal fibrosis (blue arrow) and focal aggregation of neoplastic cells (green arrow) (scale bare:50 µm). (C) Liver of tumor bearing mice treated only with laser irradiation showing birding portal inflammation (White circle) (scale bare:50 µm).). (D) Liver of tumor bearing mice treated only with PoSe-NPs showing birding portal inflammation (White circle) (scale bare:50 µm). (E) Liver of tumor bearing mice treated with PoSe-NPs then irradiated with laser radiation, showing no inflammatory cells and normal hepatic cells (orange arrow) could be detected (scale bare:50 µm).

FIGURE 12

Transmission electron microscope photomicrographs of liver tissue from NEAC, EAC mice treated with nano, laser, and Nano + laser. A NEAC Control group (A) showing the cytoplasm of the hepatocyte containing rounded euchromatic nucleus (N) with prominent nucleolus (Nu), oval mitochondria (M), and rough endoplasmic reticulum (RER). EAC group (B) showing hepatocytes contained nuclei with clumped dense chromatin the nucleus appeared irregular in shape (N), and slightly swollen mitochondria (M). Laser group (C) showing swollen mitochondria (M), and RER dilatation (RER). PoSeNPs group (D) showing polymorphic mitochondria (M) with dense granules, maricets, and deposition of aggregated PoSeNPs, and the nucleus appeared irregular in shape (N). PoSeNPs + laser Group (E) showing nearly normal architecture of hepatocytes. The cytoplasm contained rounded euchromatic nucleus (N) with prominent nucleolus (Nu), the mitochondria (M), and (RER) retain its normality.