2-Dimensional in vitro culture assessment of ovarian cancer cell line using cost effective silver nanoparticles from Macrotyloma uniflorum seed extracts

Abstract

Our research focused on generating AgNPs using Macrotyloma uniflorum (MU) seed extracts and studied their efficacy in combating tumor growth using the 2-Dimensional method for ovarian cancer cell line-PA-1. Characterization studies including a UV-visible spectrophotometer confirmed the surface plasmon resonance peak of 436 nm. Particle size determination data validated the nanoparticle diameter of 91.8 nm. Synthesized AgNPs possess a negative charge of -28.0 mV, which was confirmed through the zeta potential study. Structural characterization studies including XRD determined the crystal phase of AgNPs at four distant peaks at 2θ (38.17, 44.36, 64.52, and 77.46) and were assigned to 111, 200, 220, and 311 planes of the FCC. FTIR studies have confirmed the presence of O-H, N-H, C=O, ethers, C-Br, and C-I groups in AgNPs respectively. DPPH study has confirmed the presence of free radicles and we observed that at 500 μg/ml concentration, 76.08% of free radicles were formed which shows their efficiency. MTT assay shows the efficacy of MU-AgNPs in reducing the cell viability. At lower concentrations of MU-AgNP, 66% viability was observed and 9% of viability was observed at higher dose. ROS production (21%) was observed using MU-AgNPs with respect to 0.45% in controls, which affirms the capacity to induce DNA damage via apoptosis. Standard drug camptothecin generated 26% of ROS production which confirms higher potential of AgNPs in inducing DNA damage in tumor cells without causing lethality to the healthy cells. Further, the Fluorescence-activated cell sorting (FACS) study using a standard Caspase-3 marker confirms the generation of apoptotic bodies using two different concentrations of MU-AgNPs. At 40 μg, 64% of apoptotic cell death was observed, whereas, using 20 μg, 23% of apoptosis was recorded via fluorescent intensity. Propidium iodide-based Cell cycle study has shown a significant decrease in G0/G1 phase compared to control (88.8%), which further confirmed the apoptotic induction. Matrix metalloproteinases (MMP) studies using JC-1 dye, showed a significant increase in green fluorescence owing to lowered membrane potential, thus ensuring the breakdown of mitochondrial potential compared to untreated and standard drugs. With the obtained results, we are concluding that MU-AgNPs has a tremendous capacity to suppress the ovarian cancer cell proliferation in vitro by inducing DNA damage and apoptosis.

Chemotherapeutic Potential of AgNP Orchestrated Semecarpus anacardium Nut Extracts Against Ovarian Cancer Cell Line, PA-1

“…SA-AgNPs has a peak absorbance at 412 nm for 0.1mM and 427 nm for 0.2 mM after 30 minutes of incubation (Figure 3). Similar wavelength was observed for other plants including Macrotyloma uniflorum and Argyreia nervosa [18,37]. The average size of spherical nano particles within the SPR region in general is 410-450 nm.…”

“…Long term treatment using chemotherapy often results in tumor recurrence, which results in the combinational therapy treatments in recent years [46][47][48]. In traditional medicine, ovarian cancer is treated with a variety of herbs and plants, including Boerhavia erecta, Allium sativum, Fumaria officinalis, Zingiber officinale, Camellia sinensis, Ginkgo biloba, Quercus tinctoria, Nigella Sativa, Taxus brevifolia, Azadirachta indica, Malva sylvestris, Melia azedarach and Macrotyloma Uniflorum [16][17][18][19]. It is anticipated that the antiproliferative effects of metal nanoparticles on ovarian cancer cells will be substantially stronger if they are synthesized using these plants [19][20][21][22]71].…”

“…Next day, cells were treated with SA-AgNPs at a wide range of concentrations ranging from 0.1 µg/ml to 1000 µg/ml. Further steps were followed as mentioned earlier [18]. Flow Cytometry Study PA-1 cells were cultured in 6-well plates at an initial density of 2 × 10 5 cells/2ml into FACS tubes.…”

Portable Mid-Infrared Spectroscopy Combined with Chemometrics to Diagnose Fibromyalgia and Other Rheumatologic Syndromes Using Rapid Volumetric Absorptive Microsampling

“…Vibrational spectroscopy (infrared and Raman) captures characteristic chemical “fingerprints” of the molecules, including biomarkers present in the biological samples. This technology offers a high-throughput and rapid diagnosis of diseases with minimum sample preparation and is being applied in several areas of the medical field such as cancer [ 9 , 10 ], rheumatology [ 2 ], and urology [ 11 ]. Fourier-transformed infrared (FT-MIR) spectroscopy acquires the unique biochemical signals by irradiating a polychromatic light source (4000 to 650 cm −1 ) on the sample and recording the absorbance by the vibrating functional groups of the global metabolites in the sample.…”