Metabolite Profiling and Biological Activity Assessment of Casuarina equisetifolia Bark after Incorporating Gold Nanoparticles.

OBJECTIVE: Casuarina equisetifolia bark is rich in various active metabolites and selected to be studied due to limitation of the synthetic antioxidants that have adverse side effects. The present study aimed to enhance efficiency of the most effective extract by incorporating gold nanoparticles (Au-NPs). METHODS: The phytochemical and biological measurements were carried out in total methanolic extract and its successive fractions. Moreover, these measurements were assayed in the most effective extract after incorporating Au-NPs. RESULTS: The study revealed that total methanolic extract exhibited the highest biological and cytotoxic activities as compared to other fractions. Therefore, it is considered as good candidate for nano-extract preparation. The methanolic extract incorporated with Au-NPs showed higher antioxidant, scavenging and cytotoxic activities in addition to higher inhibitory effect against α-amylase activity as compared to native extract itself. To pinpoint active agents in total methanolic extract, the secondary metabolite profiling via HPLC-MS showed that 33 and 17 metabolites were annotated in the extract before and after incorporating Au-NPs, respectively. The median lethal dose (LD50) showed that gold total methanolic nano-extract is safer than total methanolic extract. CONCLUSION: This study concluded that total methanolic C. equisetifolia bark extract is a valuable bioresource to synthesize an eco-friendly Au-NPs with health-enhancing effect as antioxidant, antidiabetic and cytotoxic agents. The present study is considered as the first report on utilization of C. equisetifolia bark in synthesis of Au-NPs by mean of green nanotechnology and investigation of its biological activity in relation to its metabolite fingerprint.

In Vitro Study on Effect of Zinc Oxide Nanoparticles on the Biological Activities of Croton tiglium L. Seeds Extracts.

OBJECTIVE: Croton tiglium L. seeds were studied against colon cancer induced chemically in rats after incorporating silver nanoparticles (Ag-NPs) but the body has no the ability to discrete silver or silver ions. Therefore, the present study was designed to reveal the biological activities of different C. tiglium L. seeds extracts incorporated with zinc oxide nanoparticles (ZnO-NPs). RESULTS: It was found that C. tiglium L. seeds provided with high contents of total protein (27.43 g/100g), carbohydrate (18.29 g/100 g) and lipid (46.31 g/100 g). The chromatographic techniques revealed that concentrations of the predominant compounds increased in all studied extracts (ethanolic, aqueous and petroleum ether) after incorporating ZnO-NPs. The in vitro biological activities showed that the aqueous extract possessed the highest antioxidant and scavenging activities. It exhibited the highest inhibitory effect on α-amylase (41.89%) and acetyl cholinesterase (AChE) (23.00%) in addition to its higher anti-arthritic activity. All the biological activities increased after incorporating ZnO-NPs. It showed the highest cytotoxic activity that increased after incorporating ZnO-NPs against human colon carcinoma (CACO-2) cells. Therefore, this extract was selected for undergoing further studies on CACO-2 cells. The aqueous extract incorporated with ZnO-NPs arrested growth of CACO-2 cells at G2/M and increased percentage of total apoptotic cells and necrosis. The median lethal dose (LD50) showed that the extracts incorporated with ZnO-NPs were safer than the native extracts. CONCLUSION: The study showed that the aqueous extract was the most active extract that consequently exhibited promising biological activities after incorporating ZnO-NPs.

Assessment of the Biological Activities of Egyptian Purslane (Portulaca oleracea) Extract after Incorporating Metal Nanoparticles, in Vitro and in Vivo Study.

OBJECTIVE: Egyptian Purslane (Portulaca oleracea) is rich in omega-3 fatty acids and a wide range of vitamins and phyto-constituents that were absorbed slowly due to their high molecular weights. Therefore, this study was designed to accelerate the absorption of these phyto-constituents and hence increase their bioavailability by incorporating silver (Ag-NPs) and zinc oxide nanoparticles (ZnO-NPs) due to their impressive properties. METHODS: The major phyto-constituents and different biological activities were quantified in aqueous extract before and after incorporating metal nanoparticles (M-NPs). The efficiency of ZnO-P. nano-extract was studied on cell cycle and apoptosis of human hepatocellular carcinoma (HEPG-2) cells. Then, both Ag- and ZnO-P. nano-extracts were studied against hepatic fibrosis induced by thioacetamide (TAA) in rats through undergoing different hematological and biochemical measurements in addition to the histopathological examination in hepatic tissues compared to the extract itself. RESULTS: The ZnO-P. nano-extract showed significantly (P<0.05) higher antioxidant and scavenging activity due to the existence of higher total polyphenolic content. Also, it exhibited a significantly (P<0.05) higher inhibitory effect on acetyl cholinesterase (AChE) activity and higher cytotoxic activity against HEPG-2 cells. Therefore, ZnO-P. nano-extract was studied against the cell cycle and apoptosis of HEPG-2 cells compared to the extract itself. It was found that ZnO-P. nano-extract was safer than Ag-P. nano-extract. Both Ag- and ZnO- P. nano-extracts were studied against the hepatic fibrosis induced by thioacetamide (TAA) compared to the native extract. It was noticed that ZnO-P. nano-extract exhibited an ameliorative effect against hepatic fibrosis by decreasing levels of inflammatory and fibrotic markers significantly (P<0.05) more than Ag-P. nano-extract. Furthermore, it improved the antioxidant status of the hepatic tissue in addition to restoring the histopathological architecture of liver tissue. CONCLUSION: ZnO-P. nano-extract showed higher in vitro and in vivo biological activities than Ag-P. nano-extract and native P. extract itself.

Evaluation of the Biological Efficiency of Silver Nanoparticles Biosynthesized Using Croton tiglium L. Seeds Extract against Azoxymethane Induced Colon Cancer in Rats.

BACKGROUND: Colorectal cancer (CRC) is considered as the most common type of gastrointestinal cancers. Chemotherapy became limited due to the adverse side effects. Therefore, the most effective Croton tiglium extract was selected to be incorporated by silver nanoparticles (Ag-NPs) then evaluated against colon cancer induced by azoxymethane (AOM) in rats. METHODS: Different hematological and biochemical measurements were quantified in addition to markers of oxidative stress. Specific tumor and inflammatory markers were assayed. Colonic tissues were examined histopathologically in addition to immunohistochemistry (IHC). Native proteins and isoenzymes patterns were electrophoretically assayed beside expression of Tumor Protein P53 (TP53) and Adenomatous Polyposis Coli (APC) genes in colonic tissues. RESULTS: It was found that AOM caused significant (P≤0.05) elevation in the hematological and biochemical measurements. C. tiglium nano-extract restored these measurements to normalcy. Tumor and inflammatory markers elevated significantly (P≤0.05) in sera of AOM induced colon cancer group in addition to increasing peroxidation products with decline in antioxidant enzymes activities in colon tissues. Nano-extract restored these measurements to normalcy in post-treated group. Histopathological study revealed that nano-extract minimized severity of inflammatory reactions in all nano-extract treated groups and prevented  anti-Keratin 20 antibody expression in post-treated group. The lowest similarity index (SI%) values were noticed with electrophoretic protein (SI=71.43%), lipid (SI=0.00%) and calcium (SI=75.00%) moieties of protein patterns, catalase (SI=85.71%), peroxidase (SI=85.71%), α-esterase (SI=50.00%) and ß-esterase (SI=50.00%) isoenzymes in colon cancer group. Furthermore, AOM altered the relative quantities of total native bands. The nano-extract prevented the alterations that occurred qualitatively in nano-extract post-treated group and quantitatively in all nano-extract treated groups. Levels of TP53 and APC gene expression increased in AOM injected group and nano-extract restored their levels to normalcy in the post-treated group. CONCLUSION: C. tiglium nano-extract exhibited ameliorative effect against the biochemical and molecular alterations induced by AOM in nano-extract post-treated group.