TITLE:
Anti-Angiogenic and Anti-Metastatic Effects of Biogenic Silver Nanoparticles Synthesized Using Azadirachta indica
AUTHORS:
Shedrack Reuben Kitimu, Peter Kirira, Ahmed A. Abdille, Judith Sokei, Dominic Ochwang’i, Peter Mwitari, Andrew Makanya, Naomi Maina
KEYWORDS:
Biogenic AgNPs, Cancer, Angiogenesis, Metastasis, CAM
JOURNAL NAME:
Advances in Bioscience and Biotechnology,
Vol.13 No.4,
April
15,
2022
ABSTRACT: Background: Nanotechnology symbolizes a broad discipline with enormous potential in
cancer treatment bridging one of the bottlenecks of traditional approaches in
cancer therapy which is an inability to deliver adequate quantities of
anti-cancer drugs to the tumor area. Studies on nanoparticles indicate their
importance in cancer angiogenesis and metastasis. Aim: The present study
assessed anti-angiogenesis and anti-metastatic effects of biogenic silver
nanoparticles (AgNPs) synthesized from neem plant (Azadirachta indica). Methods: Chicken
chorioallantoic membrane (CAM) and two-dimensional (2D) wound healing assays
were used to study anti-angiogenic and anti-metastatic effects of the AgNPs
respectively. Twenty-four fertilized eggs were divided into four groups: two
biogenic AgNPs treatments at 100 μg/ml and 200 μg/ml; negative control (1%
DMSO) and positive control (cyclophosphamide). On day 8 of incubation, filter
discs impregnated with different concentration levels of the treatments were
placed on the CAM. On day 12 of incubation, the CAMs were imaged using a
stereomicroscope, scaled using ImageJ, and different morphometric and spatial
parameters computed using AngioTool software. Vessel area, vessel percent area,
total number of junctions, total vessel length, average vessel length, mean
lacunarity, and junction density were measured. The crown-rump length (CRL) and
fetal weight were also recorded on day 16 of incubation. In order to determine
relative gene expression profiles of iNOS and VEGF, total RNA was extracted
from the CAM, and qRT-PCR was performed
with β-Actin as a reference gene. For
the 2D wound healing assay, DU145 human prostate cells were grown in Dulbecco’s
Modified Eagle’s Medium supplemented with 10% Fetal Bovine Serum. Results: Biogenic AgNPs demonstrated anti-angiogenic effects in a dose-dependent manner
in the parameters generated from the CAM images. Also, qRT-PCR revealed
down-regulation of iNOS and VEGF genes. The 2-dimensional wound healing assay
showed inhibition of migration and motility of the DU145 cells for the 72-hours
of assessment. Conclusion: The present study postulates that the
biogenic AgNPs can prevent angiogenesis by inactivation of VEGF-NO and
VEGF/VEGF-R pathways while inhibiting cell migration and metastasis.