In Situ Bio-responsive Biosynthesis Of Ag-Fe₃O₄ Nanoclusters In Exosomes For Early Cancer Diagnosis

Accurate cancer recognition and diagnosis is essential for its effective therapy.Nowadays,it is still desirable to find efficient ways for the early diagnosis and treatment of cancers.Especially,it continues attracting much attention to find the biocompatible and precise imaging approaches for accurately detecting cancer locations.Our recent studies illustrate that the biosynthesis of functional nanoclusters as effective biomarkers in redox-imbalanced mammalian cancer cells is effortless,cost-efficient,and biologically friendly with advanced biocompatibility,mark clarifies,and reduces inauspicious influence compared with conventional simulated access.Meanwhile,exosomes,the extracellular vesicles that were released from all types of cells,usually have a diameter range of less than 40-150 nm and have currently achieved enormous attention in the scientific fraternity as therapeutic vehicles and potential diagnostic biomarkers for the diverse disorders,including cancers.The exosomes texture is basically depending upon the category and environment of the cells where released,which were reported as numerous cargos of proteins,mi RNAs,m RNA,si RNA,and numerous cellular organelles as well as mitochondria,etc.Based on these observations,in this study,we have explored a new strategy to utilize cancer cells including Hep G2 to in situ bio-responsive self-assembly biosynthesize Ag-Fe₃O₄ nanoclusters in exosomes for early cancer diagnosis.1)The biosynthesized fluorescent and magnetic Ag-Fe₃O₄nanoclusters（NCs）in exosomes were realized in situ in cancer cells by using their respective salts（i.e.,Ag NO₃ and Fe Cl₂,respectively）in the presence of glutathione（GSH）.The intracellular existence of the biosynthesized Ag-Fe₃O₄NCs were characterized by UV-Vis absorption spectroscopy,Fourier transforms infrared（FTIR）spectra,Energy spectrum（EDS）,Scanning electron microscope（SEM）,Transmission electron microscopy（TEM）and so on.2)We have isolated the Ag-Fe₃O₄NCs loaded exosomes from cancer cells,including the Human liver hepatocellular carcinoma（Hep G2）of the treated after the in situ biosynthesis from their respective pre-ionic solutions,in addition of GSH to reduce the oxidative stress caused by the formation of the Ag-Fe₃O₄nanoclusters.The Ag-Fe₃O₄NCs loaded exosomes were then isolated and characterized by electron microscopy and confocal fluorescence microscopy studies,whereas the Ag NCs potential as fluorescence imaging probes and Fe₃O₄NCs as a contrast agent for CT and MRI.Furthermore,it is observed that these isolated exosomes from Hep G2 or U87 cancer cells significantly influence of cellular uptake and cell viability.These findings demonstrate the biocompatible nature of Ag-Fe₃O₄ NCs loaded on exosomes can be utilized to bioimage cancer at initial stages through fluorescence imaging.