Preparation of biogenic silver chloride nanoparticles from microalgae Spirulina Platensis extract: anticancer properties in MDA-MB231 breast cancer cells.

BACKGROUND: Breast carcinoma is the second leading cause of cancer related-deaths among women. Given its high incidence and mortality rates, searching for innovative treatments represents a formidable challenge within the medical and pharmaceutical industries. This study delves into the preparation, characterization, and anticancer properties of silver chloride nanoparticles (AgCLNPs) as a novel therapeutic approach for breast cancer cells, employing a biological synthesis method. METHODS: This investigation, utilized spirulina platensis extract to synthesize silver chloride nanoparticles (AgCLNPs-SP). The formation, size, and structure of the nanoparticles were characterized by Transmission Electron Microscopy (TEM), Scanning Electron Microscope (SEM), X-ray crystallography (XRD), and Energy-dispersive X-ray spectroscopy (EDS) analysis. Additionally, the apoptotic and anticancer properties of AgCLNPs-SP were thoroughly examined. RESULTS: The results, revealed AgCLNPs-SP to exhibit a spherical, morphology with a size range of 40-70 nm, primarily silver and chlorine. The dose-dependent response of AgCLNP-SP against MDA-MB231 cells was ascertained using the MTT Assay, with an IC50 value of 34 µg/mL. Furthermore, the Annexin V-FITC/ PI apoptosis assay demonstrated a significant proportion of early apoptosis (43.67%) in MDA-MB231 cells. This apoptosis process was substantiated by up-regulation in mRNA expression levels of P53, CAD, and Bax genes, alongside a down-regulation of the of bcl2 gene expression. Additionally, an augmented production of reactive oxygen species (ROS), cell cycle analysis, Hoechst staining assay, and evaluated levels of Caspase - 3, -8 and - 9 were observed in AgCLNPs-SP-treated MDA_MB231 cancer cells. CONCLUSIONS: In conclusion, the results suggest that AgCLNPs-SP may be a promising agent for treating breast cancer.

Apoptosis Induction by ZnFe2O4-Ag Biosynthesized by Chlorella vulgaris in MCF-7 Breast Cancer Cell Line.

The incidence and mortality of breast cancer are growing which indicates the inefficiency of the current chemotherapy drugs. Due to the anticancer potential of Zn and Ag and the magnetic feature of iron oxide, in this work, we synthesized ZnFe2O4-Ag nanocomposite using Chlorella vulgaris and investigated its anticancer effect on breast cancer cell line. Physicochemical characterization was performed by FT-IR, XRD, SEM, TEM, VSM, EDS mapping, UV, and zeta potential assays. Cell cytotoxicity and apoptosis frequency were studied by the MTT and flow cytometry assays. Also, cell cycle analysis, Hoechst staining, and measuring ROS (reactive oxygen species) level were performed. The synthesized particles were almost spherical with a size range of 14-52 nm. The FT-IR and XRD assays confirmed the proper synthesis of the particles and VSM analysis showed that particles had magnetic property and the maximum saturation magnetization was 0.8 Emu/g. Also, the EDS mapping of the nanocomposite showed the Zn, Fe, O, and Ag elements. The MTT assay showed that the 50% inhibitory concentration (IC50) of ZnFe2O4-Ag for breast cancer and normal cells were 28 and 154 µg/mL, respectively, and the nanocomposite had stronger anticancer activity than cisplatin (IC50 = 84 µg/mL). Flow cytometry analysis showed that the exposure to the nanocomposite induced cell apoptosis by 77.5% and significantly induced ROS generation. Also, treating breast cancer cells with the nanocomposite induced cell cycle arrest and apoptotic features, including chromatin condensation and fragmentation. In conclusion, ZnFe2O4-Ag nanocomposite synthesized by C. vulgaris could suppress the proliferation of breast cancer cells by the generation of oxidative stress, apoptosis induction, and cell cycle arrest.

Antineoplastic effectiveness of silver nanoparticles synthesized from Onopordum acanthium L. extract (AgNPs-OAL) toward MDA-MB231 breast cancer cells.

BACKGROUND: The present research was done to investigate the anticancer properties of silver nanoparticles (AgNPs) fabricated using bioactive extract of Onopordum acanthium L. (AgNPs-OAL) against breast cancer cells MDA_MB231 in vitro. METHODS: The determination studies of AgNPs-OAL were confirmed by X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM) analysis. Interestingly, the FESEM image observed the spherical shape of AgNPs-OAL with the range of 1-100 nm. RESULTS: As AgNP-OAL exhibited significant cytotoxicity properties on breast cancer MDA_MB231 cells with IC50 values of 66.04 µg/mL, while lowing toxicity toward normal human embryonic kidney 293 (HEK293) cells with IC50 values of 101.04 µg/mL was evaluated. Further, up-regulation of apoptotic Bax and CAD gene expressions were confirmed by quantitative real-time reverse transcription-PCR (qRT-PCR) technique results. Moreover, enhanced cell cycle population (sub-G1), annexin V/PI staining, acridine orange and ethidium bromide (AO/EB) staining, Hoescht 33,258 dye, and generation of reactive oxygen species were observed in AgNP-OAL-treated MDA_MB231 cancer cells. CONCLUSIONS: The green-synthesized AgNP-OAL has promising anticancer efficiency that can trigger apoptosis pathways in the MDA_MB231 breast cancer cells.

Preparation, characterization, and anticancer efficacy of novel cobalt oxide nanoparticles conjugated with thiosemicarbazide.

Gastric cancer is one of the most common cancers in modern societies. Previous studies have shown that the use of nanoparticle complexes is effective in the treatment of cancer. The aim of this study was to investigate the cytotoxicity and anticancer properties of cobalt oxide (Co3O4) nanoparticles (NPs) functionalized by glutamic acid (Glu) and conjugated with thiosemicarbazide (TSC) on gastric cancer (AGS) cell line. First, the Co3O4@Glu/TSC nanoparticles were synthesized via co-condensation reaction. Fourier-transform infrared (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM) and energy dispersive X-ray (EDX) tests were performed for identifying the morphology, structure, size and functional groups of produced nanoparticles. MTT assay was also performed to evaluate cytotoxicity effect. Moreover, Annexin V/PI staining with flow cytometry analysis, caspase-3 activation assay, and Hoechst 33258 staining was carried out for evaluating apoptosis. The FTIR results showed that the components of Co3O4@Glu/TSC NPs complex were successfully fabricated. Crystallographic structure of Co3O4@Glu/TSC NPs was confirmed by XRD patterns. SEM results indicated that the size of the nanoparticles was in the range of 16-40 nm. An EDX spectrum was determined and data explained the existence of cobalt as the prominent element. MTT test results showed that AGS cell life was significantly decreased compared to the control group with increasing concentration of nanoparticles (dose-dependent) (P < 0.05), IC50 = 107.5 µg/mL. The results of flow cytometry assay and caspase-3 activity showed that fabricated Co3O4@Glu/TSC NPs induced apoptosis in the treated group. Moreover, Co3O4@Glu/TSC NPs treated AGS cells indicate an increase in the apoptotic characteristics including nuclear fragmentation. In the current work, the promising cytotoxicity and anti-cancer activities of Co3O4@Glu/TSC NPs complex toward gastric cancer (AGS) cell line were showed and it can be suggested for the drug delivery system.

Zinc-Phosphate Nanoparticles as a Novel Anticancer Agent: An In Vitro Evaluation of Their Ability to Induce Apoptosis.

In the current study, zinc-phosphate nanoparticles (ZnPNPs) were investigated for the first time due to their anticancer activity against breast cancer Michigan Cancer Foundation-7 (MCF-7) cell line. The modification of such nanoparticles (NPs) was further examined for physicochemical characterization using various techniques such as powder X-ray diffraction (XRD), dynamic light scattering (DLS), zeta potential calculation, field emission scanning electron microscopy (FESEM), energy-dispersed spectroscopy (EDS), and Fourier transform infrared (FTIR) spectroscopy. Then, the newly fabricated ZnPNPs were tested for their in vitro cell cytotoxicity against breast cancer MCF-7 cells and noncancerous human embryonic kidney HEK293 cells, using MTT assay as a colorimetric one to assess cell metabolic activity for 24 h. The apoptotic efficacy of the NPs was subsequently confirmed through data obtained from Annexin V-fluorescein isothiocyanate (FITC)/propidium iodide (PI) staining kit and cell cycle analysis. Determination of reactive oxygen species (ROS) generation was further performed via flow cytometry. Additionally, the expression of tumor suppressor genes p53 was analyzed using real-time polymerase chain reaction (PCR). Also, the prepared NPs showed a mean particle size of 38 nm. The measurements correspondingly showed that the cytotoxicity of MCF-7 cells depends on the concentration of NPs (IC50 = 80.112 µg/mL). MCF-7 cells were associated with initiation of apoptotic pathway in cells. Additionally, flow cytometry revealed cell cycle arrest in sub-G1 phase. ROS production was also obtained after treatment with IC50 concentration. According to annexin V-FITC/PI staining kit data, the percentage of early and late apoptotic cells was 78.2% in those treated with ZnPNPs. Moreover, the real-time PCR results demonstrated the ability of NPs in upregulating p53 gene expression. In summary, the data demonstrated that fabricated ZnPNPs had prominence to act as antitumor agents in breast cancer therapy.

Novel pyridinecarboxaldehyde thiosemicarbazone conjugated magnetite nanoparticulates (MNPs) promote apoptosis in human lung cancer A549 cells.

The present study highlights the apoptotic activity of magnetic Fe3O4 nanoparticulates functionalized by glutamic acid and 2-pyridinecarboxaldehyde thiosemicarbazone (PTSC) toward human lung epithelial carcinoma A549 cell line. To this aim, the Fe3O4 nanoparticulates were prepared using co-precipitation method. Then, the glutamic acid and Fe3O4 nanoparticulates were conjugated to each other. The product was further functionalized with bio-reactive PTSC moiety. In addition, the synthesized Fe3O4@Glu/PTSC nanoparticulates were characterized by physico-chemical techniques including scanning electron microscope (SEM), energy dispersive X-ray (EDX), X-ray diffraction (XRD), Fourier-transform infrared (FT-IR) spectroscopy and zeta potential analysis. The effects of in vitro cell viability in Fe3O4@Glu/PTSC nanoparticulate indicated the anti-proliferative properties in a dose-dependent manner (IC50 = 135.6 µM/mL). The high selectivity for tumor cells and far below of activity in HEK293 non-tumorigenic cells is considered as an important feature for this complex (SI, 3.48). Based on the results, PTSC failed to reveal any activity against A549 cells alone. However, Fe3O4 nanoparticulates had some effects in inhibiting the growth of lung cancer cell. Furthermore, Bax and Bcl-2 gene expressions were quantified by real-time PCR method. The expression of Bax increased 1.62-fold, while the expression of Bcl-2 decreased 0.76-fold at 135.6 µM/mL concentration of Fe3O4@Glu/PTSC compared to untreated A549 cells. Furthermore, the Fe3O4@Glu/PTSC nanoparticulate-inducing apoptosis properties were evaluated by Hoechst 33258 staining, Caspase-3 activation assay and Annexin V/propidium iodide staining. The results of the present study suggest that Fe3O4@Glu/PTSC nanoparticulates exhibit effective anti-cancer activity against lung cancer cells.

Green Engineered Biomolecule-Capped Silver Nanoparticles Fabricated from Cichorium intybus Extract: In Vitro Assessment on Apoptosis Properties Toward Human Breast Cancer (MCF-7) Cells.

The current experiment reveals the anticancer properties of silver nanoparticles (AgNPs) synthesized using aqueous leaf extract of Cichorium intybus, a significant medicinal plant. The characteristics of AgNPs were continuously studied by powder X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), zeta potential, transmission electron microscopy (TEM), scanning electron microscopy (SEM), and energy-dispersive spectroscopy (EDS) analysis. Current microscopic results show that produced AgNPs were spherical in shape with an average size of 17.17 nm. A strong peak between 2 and 4 keV showed the greatest ratio of the elemental silver signals, due to surface plasmon resonance (SPR). The AgNPs, fabricated by green method, had a negative zeta potential of - 9.76 mV, which indicates that the synthesized AgNPs is dispersed in the medium with high stability. The in vitro cytotoxicity effect of AgNPs showed promising anticancer activity against human breast cancer MCF-7 cells. Annexin V-FITC/propidium iodide assay, Hoechst 33258 staining, and upregulation of caspase 3 activity revealed significant apoptosis activities of AgNPs against MCF-7 cells. Moreover, the flow cytometric analyses of cell cycle distribution of MCF7 cells showed that AgNPs treatment has enhanced the sub-G1 peaks, which is an indicator of apoptosis pathway. Overall results in our study suggested that AgNPs fabricated by a biogreen approach could be useful in cancer therapy.

Impact of a Missense Variation (p.S150R:AGC>AGG) in the XRCC2 Gene on Susceptibility to Colorectal Cancer.

BACKGROUND: The X-ray repair complementing defective repair in Chinese hamster cells 2 (XRCC2) is an important protein in response to DNA double-strand breaks (DSBs) in human cells. XRCC2, as a functional protein in the homologous recombination repair (HRR) process, has been identified to have several polymorphisms which might be associated with the risk of cancer. Therefore, we aimed to investigate a novel missense variation (AGC>AGG, p.Ser150Arg) in the XRCC2 gene for colorectal cancer susceptibility. METHODS: We studied 291 colorectal cancer (CRC) patients and 140 healthy individuals. ARMS PCR method was used to detect the AGC>AGG (p.Ser150Arg) variation in the XRCC2 gene. RESULTS: The results showed that there was a significant differential among CRC and controls in the genotypic and allelic frequencies (p < 0.001) of XRCC2; AGC>AGG, p.Ser150Arg. Our results demonstrated that the G allele of XRCC2; AGC>AGG, p.Ser150Arg was associated with increased CRC risk (odds ratio = 59.04, 95% confidence interval = 18.6 - 186). This variation also influenced CRC cancer susceptibility in smokers (p < 0.001). CONCLUSIONS: The G allele of XRCC2; AGC>AGG, p.Ser150Arg, may be a potential marker for CRC in Iranians and investigations in other populations are warranted for further universal application in CRC detection and prediction.

Anti-cancer and anti-oxidant properties of ethanolic leaf extract of Thymus vulgaris and its bio-functionalized silver nanoparticles.

This study highlights the anti-oxidant and anti-cancer activities of bio-functionalized Thymus vulgaris silver nanoparticles (TVAgNPs) and bioactive compounds were compared using the human breast cancer T47D cell line. The aqueous ethanolic extract of T. vulgaris evaluated for chemical composition using the gas chromatography-mass spectrometer (GC-MS) analysis. The prepared TVAgNPs were determined by means of UV-Vis spectroscopy, FTIR spectroscopy, zeta potential, scanning electron microscopy, transmission electron microscopy, and energy-dispersed spectroscopy analysis. The T. vulgaris extract and TVAgNPs were studied for their in vitro anti-oxidant property by 2, 2-diphenyl, 1-picryl hydrazyl (DPPH) assay. Microscopic observations indicated spherical shaped and monodispersed nanoparticles and the average size of the nanoparticles was about 30 nm. Regarding the elemental composition profile of the TVAgNPs, the highest signal of silver (89.30%) was detected followed by other elements. An absorption peak was registered at 440 nm according to surface plasmon resonance (SPR) of the TVAgNPs in solution. A zeta potential of fabricated nanoparticles was approximately - 12.6 mV, indicating higher stability of the bio-functionalized TVAgNPs. The T. vulgaris extract and synthesized TVAgNPs were evaluated for their anti-cancer activity using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay and Annexin V double staining with propidium iodide (PI) flow cytometric analysis toward T47D cells. The cytotoxicity properties of the bio-functionalized T. vulgaris AgNPs revealed that the sensitivity of T47D human breast cancer cells is high compared with T. vulgaris extract. The Annexin V/PI staining indicated that the fabricated TVAgNPs shows increased apoptosis in T47D cells as compared to untreated cells. Besides, the anti-oxidant activity of the TVAgNPs clarified a higher anti-radical-scavenging activity compared to Thymus vulgaris extract. Our data show that the potential biological activities of the bioactive constituents of T. vulgaris can be enhanced through bio-functionalized T. vulgaris AgNPs due to the bioorganic compounds that exist in the extract.

Gd3+-Asparagine-Anionic Linear Globular Dendrimer Second-Generation G2 Complexes: Novel Nanobiohybrid Theranostics.

Designing a unique theranostic biocompatible, biodegradable, and cost-effective agent which is easy to be synthesized as a biohybrid material was the aim of this study. In this matter, asparagine attached to anionic linear globular dendrimer G2 (as a biocompatible, biodegradable, and cost-effective agent which is negatively charged nanosized and water soluble polymer that outweighs other traditionally used dendrimers) and finally contrast agent (Gd3+) was loaded (which made complexes) in synthesized asparagine-dendrimer. Observations revealed that, in addition to successful colon cancer and brain targeting, Gd3+-dendrimer-asparagine, the proposed theranostic agent, could increase T1 MR relaxation times, decrease T2 MR relaxation times significantly, and improve contrast of image as well as illustrating good cellular uptake based on florescent microscopy/flow cytometry and ICP-mass data. In addition to that, it increased tumor growth inhibition percentage (TGI%) significantly compared to FDA approved contrast agent, Magnevist. Totally, Gd3+-anionic linear globular dendrimer G2-asparagine could be introduced to the cancer imaging/therapy (theranostics) protocols after in vivo MR and fluorescent analysis and passing clinical trials. Hence, this nanotheranostic agent would be a promising candidate for brain drug delivery and imaging in the future.

Antioxidant, antibacterial and anticancer properties of phyto-synthesised Artemisia quttensis Podlech extract mediated AgNPs.

The focus of this study is on a rapid and cost-effective approach for the synthesis of silver nanoparticles (AgNPs) using Artemisia quttensis Podlech aerial parts extract and assessment of their antioxidant, antibacterial and anticancer activities. The prepared AgNPs were determined by ultraviolet-visible spectroscopy, X-ray diffraction, Fourier transform infra-red spectroscopy, transmission electron microscopy, scanning electron microscopy, energy-dispersive spectroscopy, and dynamic light scattering and zeta-potential analysis. The AgNPs and A. quttensis extract were evaluated for their antiradical scavenging activity by 2, 2-diphenyl, 1-picryl hydrazyl assay and anticancer activity against colon cancer (human colorectal adenocarcinoma cell line 29) compared with normal human embryonic kidney (HEK293) cells. Also, the prepared AgNPs were studied for its antibacterial activity. The AgNPs revealed a higher antioxidant activity compared with A. quttensis extract alone. The phyto-synthesised AgNPs and A. quttensis extract showed a dose-response cytotoxicity effect against HT29 and HEK293 cells. As evidenced by Annexin V/propidium iodide staining, the number of apoptotic HT29 cells was significantly enhanced, following treatment with AgNPs as compared with untreated cells. Besides, the antibacterial property of the AgNPs indicated a significant effect against the selected pathogenic bacteria. These present obtained results show the potential applications of phyto-synthesised AgNPs using A. quttensis aerial parts extract.

Novel imatinib-loaded silver nanoparticles for enhanced apoptosis of human breast cancer MCF-7 cells.

In the current study, in vitro biological feature of imatinib-loaded silver nanoparticles (IMAB-AgNPs) on human breast cancer cell line was investigated. The formation of synthesized silver nanoparticles (AgNPs) was characterized by UV-Visible spectroscopy, EDS, TEM imaging, SEM, FTIR, DLS and Zeta potentiometer. The developed IMAB-AgNPs with maximum percentage of loading efficiency was demonstrated in the average of 130 nm and mostly spherical. Additionally, in vitro drug release study showed a slow and continuous release of imatinib over a period of 80 h. We demonstrated that the synthesized IMAB-AgNPs exhibited a dose-dependent cytotoxicity against MCF-7 cell line. Then, real-time PCR method was also applied for the investigation of Bax and Bcl-2 gene expression in the cells. Comparing IMAB-AgNPs to AgNPs and Imatinib revealed the ability of IMAB-AgNPs to up-regulating Bax/Bcl-2 ratio. An induction of apoptosis was evidenced by Annexin-V/PI detection assay. Based on the current obtained data, the IMAB-AgNPs can exhibit inhibitory effect on viability through up regulation of apoptosis in MCF-7 cancer cells, which provides influencing evidence for the green synthesized AgNPs as a promising sustained drug delivery system.

Chemical composition, antioxidant, antibacterial and cytotoxic effects of Artemisia marschalliana Sprengel extract.

The present study was to investigate the gas chromatography/mass spectrometry (GC/MS), in vitro antioxidant, antibacterial and anticancer activity of the ethanolic extract from aerial parts of Artemisia marschalliana Sprengel against human gastric carcinoma (AGS) and L929 cell lines. Phytochemical analysis of A. marschalliana Sprengel extract showed 22 major components and the most dominant compounds were trans-phytol (29.22%), α-Linolenic acid (13.47%) and n-Hexadecanoic acid (9.28%). In addition, the antioxidant and anticancer activity of A. marschalliana Sprengel extract were evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) methods, respectively. Antibacterial activity against selected pathogenic bacteria was also determined. According to the present obtained results, it seems that this plant has potential uses for pharmaceutical industries and further studies of pharmaceutical importance were suggested to be performed on A. marschalliana Sprengel.

Green synthesis and evaluation of silver nanoparticles as adjuvant in rabies veterinary vaccine.

BACKGROUND: Green synthesis of nanoparticles by plant extracts plays a significant role in different applications. Recently, several studies were conducted on the use of nanoparticles as adjuvant. The main aim of this study was to evaluate green synthesized silver nanoparticles (AgNPs) as adjuvant in rabies veterinary vaccine and compare the results with the existing commercially available alum adjuvant. MATERIALS AND METHODS: In the current study, AgNPs were prepared by the reduction of aqueous silver nitrate by leaf extract of Eucalyptus procera. The formation of AgNPs was confirmed by ultraviolet (UV)-visible spectrophotometer, scanning electron microscopy, dynamic light scattering, and X-ray diffraction analysis. Then, different amounts of AgNPs (200 µg, 400 µg, 600 µg, and 800 µg) were added to 1 mL of inactivated rabies virus. The loaded vaccines (0.5 mL) were injected intraperitoneally into six Naval Medical Research Institute mice in each group on days 1 and 7. On the 15th day, the mice were intracerebrally challenged with 0.03 mL of challenge rabies virus (challenge virus strain-11, 20 lethal dose [20 LD50]), and after the latency period of rabies disease in mice (5 days), the mice were monitored for 21 days. Neutralizing antibodies against rabies virus were also investigated using the rapid fluorescent focus inhibition test method. The National Institutes of Health test was performed to determine the potency of optimum concentration of AgNPs as adjuvant. In vitro toxicity of AgNPs was assessed in L929 cell line using MTT assay. In addition, in vivo toxicity of AgNPs and AgNPs-loaded vaccine was investigated according to the European Pharmacopeia 8.0. RESULTS: AgNPs were successfully synthesized, and the identity was confirmed by UV-visible spectrophotometry and X-ray diffraction analysis. The prepared AgNPs were spherical in shape, with an average size of 60 nm and a negative zeta potential of -14 mV as determined by dynamic light scattering technique. The highest percentage of viability was observed at 15 mg/kg and 20 mg/kg of AgNPs-loaded vaccine concentrations after injecting into the mice. The calculated potencies for alum-containing vaccine and AgNPs-loaded vaccine (dose 15 mg/kg) were 1.897 and 1.303, respectively. MTT assay demonstrated that alum at the concentration of 10 mg/mL was toxic, but AgNPs were not toxic. The in vivo toxicity also elucidated the safety of AgNPs and AgNPs-loaded vaccine in mice and dogs, respectively. CONCLUSION: In the current study, for the first time, the adjuvanticity effect of green synthesized AgNPs on veterinary rabies vaccine potency with no in vivo toxicity was elucidated according to the European Pharmacopeia 8.0.