GREEN SYNTHESIS OF NANOPARTICLES CONTAINING SILVER AND A FRACTION OF THE ALGAE Spatoglossum schröederi COMPOSED BY ALGINIC ACID AND FUCAN A: PHYSICAL-CHEMICAL CHARACTERIZATION AND EVALUATION OF ITS ANTIPROLIFERATIVE ACTIVITY AGAINST MELANOMA CELLS (B16F10)
Brown Algae, Acid Polysaccharides, Nanotechnology, Antitumor compounds.
Silver nanoparticles (AgNPs) have several biomedical applications, among them the antiproliferative potential. Numerous types of synthesis of silver nanoparticles are found in the literature, however, a type of synthesis less aggressive and more economically viable, a green synthesisl, has gained value among the others. In this synthesis process, it is necessary to use a reducing agent and stabilizer which are non-toxic to animal cells and to the environment. In this context, an association of biomolecules of marine origin with biological activities already described, such as the polysaccharide acids of algae, to the AgNPs gain importance. Many acidic polysaccharides (PA) extracted from marine algae are distinguished by their different activities, such as the algae Spatoglossum schröederi. These fucans have antioxidant, antiangiogenic, antitumor activity, among others. Due to the few reports of AgNPs synthesis involving algae polysaccharides, the objective of this work was to synthesize AgNPs with a rich fraction of alginic acid and fucan A from the S. schröederi algae by a green synthesis process and to test the AgNPs obtained on the lineage of melanoma B16F10. The PA of the S. schröederi algae was extracted by a proteolytic digestion process and fractionated with acetone. The higher yield fraction rich in alginic acid and fucan A, F0.5 v, was used for synthesis of the AgNPs. The characterization of the AgNPs obtained contemplated an analysis of UV light absorption, chemical composition, average diameter, polydispersity, zeta potential, infrared spectroscopy (FTIR), dispersive energy spectroscopy (EDS), scanning electron microscopy (SEM) and atomic force microscopy (AFM). The analyzes by UV-visible spectroscopy in the range between 400 and 440 nm confirmed the formation of AgNPs, they presented 196 ± 13 nm diameter, polydispersion values below 0.4 and negative zeta potential. The chemical dosages, FTIR and EDS revealed that most AgNPs are composed of acidic polysaccharides. The images obtained by SEM and AFM indicated a spherical format for AgNPs. The antiproliferative activity was evaluated by the MTT method against B16F10 cell lines and 3T3 fibroblasts, and by flow cytometry, its action on death and cell cycle was investigated. The AgNPs affected the ability to reduce MTT from the 3T3 and B16F10 lineages, but these were evidenced for a B16F10, the AgNPs (0.5 mg/mL) were able to reduce by around 50%, while a silver ion and F0.5 v at any of the concentrations influenced the reduction of MTT. Flow cytometry analyzes indicated a high labeling rate for annexin V and propidium iodide for the B16F10 cells exposed to AgNPs, in addition to an increase in the percentage of cells in SubG1 in detriment to the other phases. Additional studies are needed to fully elucidate the mechanism of antiproliferative action of AgNPs and to discover other properties of the fraction that are enhanced by the synthesis of nanoparticles. However, it can be concluded that it is possible to synthesize AgNPs with acidic polysaccharides from S. schröederi using a green method and that these AgNPs showed more pronounced antiproliferative activity than their precursors.