Mycosynthesis of selenium nanoparticles using Penicillium tardochrysogenum as a therapeutic agent and their combination with infrared irradiation against Ehrlich carcinoma

Abstract

Over the past years, the assessment of myco-fabricated selenium nanoparticles (SeNPs) properties, is still in its infancy. Herein, we have highly stable myco-synthesized SeNPs using molecularly identified soil-isolated fungus; Penicillium tardochrysogenum OR059437; (PeSeNPs) were clarified via TEM, EDX, UV-Vis spectrophotometer, FTIR and zeta potential. The therapeutic efficacy profile will be determined, these crystalline PeSeNPs were examined for antioxidant, antimicrobial, MIC, and anticancer potentials, indicating that, PeSeNPs have antioxidant activity of (IC₅₀, 109.11 μg/mL) using DPPH free radical scavenging assay. Also, PeSeNPs possess antimicrobial potential against Penicillium italicum RCMB 001,018 (1) IMI 193,019, Methicillin-Resistant Staphylococcus aureus (MRSA) ATCC 4330 and Porphyromonas gingivalis RCMB 022,001 (1) EMCC 1699; with I.Z. diameters and MIC; 16 ± 0.5 mm and MIC 500 µg/ml, 11.9 ± 0.6 mm, 500 µg/ml and 15.9±0.6 mm, 1000 µg/ml, respectively. Additionally, TEM micrographs were taken for P. italicum treated with PeSeNPs, demonstrating the destruction of hyphal membrane and internal organelles integrity, pores formation, and cell death. PeSeNP alone in vivo and combined with a near-infrared physiotherapy lamp with an energy intensity of 140 mW/cm² showed a strong therapeutic effect against cancer cells. Thus, PeSeNPs represent anticancer agents and a suitable photothermal option for treating different kinds of cancer cells with lower toxicity and higher efficiency than normal cells. The combination therapy showed a very large and significant reduction in tumor volume, the tumor cells showed large necrosis, shrank, and disappeared. There was also improvement in liver ultrastructure, liver enzymes, and histology, as well as renal function, urea, and creatinine.

Figure 1

Neighbor-joining phylogenetic tree of P. tardochrysogenum OR059437and related fungi.

Figure 2

(A) UV- Visible spectrum of myco-synthesized Se-NPs using P. tardochrysogenum OR059437, (the red circle indicated the SeNPs formation range), (B): TEM image of myco-synthesized Se-NPs of scale 600 nm, (C): Zeta potential distribution of myco-synthesized Se-NPs.

Figure 3

(A): EDX analysis of PeSeNPS, (B): Field of EDX analysis of PeSeNPs, (C): FT-IR spectrum of PeSeNPs.

Figure 4

DPPH scavenging activity of PeSeNPs.

Figure 5

Transmission Electron microscopy images showing morphological alterations in Penicillium italicum RCMB 001,018 (1) IMI 193,019 hypha treated by myco-synthesized PeSeNPs. (A) showed the untreated Penicillium italicum RCMB 001,018 (1) IMI 193,019 exhibit a compact cell wall, continuous cytoplasmic membrane, homogeneous and electron-dense cytoplasm. (B) by contrast, the effect of treating P. italicum species with our myco- synthesized PeSeNPs; an obvious destruction and detachment of membrane and internal organelles integrity, subsequently, changes in hyphal shape formation of pores and cell death.

Figure 6

Effect of different doses of PeSeNPs on EAC cell viability assessed using the MTT assay, which indicates the cytotoxic activity against EAC cells with 50% cell cytotoxic concentration (IC₅₀ = 225 ± 0.56 µg/ml).

Figure 7

Cytotoxicity of PeSeNPS toward EAC. (A) The control EAC without PeSeNPS, and (B) EAC at a 225 µg/mL concentration.

Figure 8

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

Figure 9

Survival rate of the mice after different treatment.

Figure 10

Histological analysis of tumor stained with hematoxylin and eosin (Scale bare: 50 µm). (A) Non treated tumor which shows inflammatory cells (green arrow) (B) Tumor irradiated with infrared which shows shrink of carcinoma cells (white circle) (C) injected tumor with Se-NPs which shows the beginning of cell necrosis (orange arrow circle) (D) Tumor injected with Se-NPs then exposed to infrared radiation which showing necrosis cells (blue arrow).

Figure 11

Photomicrograph of liver section of negative control group NEAC showing normal architecture of the liver, central vein (CV), and blood sinusoids (BS) (H. & E., 400X) (A). The liver section of the group with untreated tumor showed degenerated hepatocytes with congested central vein (CV), leucocytes infiltration (LI) (H. & E., 400X) (B). Liver section of group irradiated tumor with infrared which shows leucocytes infiltration (LI) (H. & E., 400X) (C). Liver section of group injected tumor with SeNPs which showed bile ductules (BD) and vacuolated hepatocytes with pyknotic (P) nuclei congested central vein (CV) (H. & E., 400X) (D). The liver section of the group injected tumor with SeNPs then irradiated by infrared light showing normal architecture of the liver, central vein (CV), and blood sinusoids (BS) (H. & E., 400X) (E).

Figure 12

Photomicrograph of Kidney section of negative control group (A). The kidney section of group with untreated tumor (B), irradiated tumor with infrared (C), injected tumor with Se-NPs (D), and injected the tumor with Se-NPs then irradiated by infrared light (E). All slides were stain by (H & E., 400X).

Figure 13

Transmission electron microscope photomicrographs of liver tissue from NEAC, EAC mice treated with, infrared radiation, PeSe-NPs, and PeSe-NPs + infrared radiation.