Previously, mucus of some molluscs has been studied as a potential source of new natural compounds capable of inducing cell proliferation and of remodelling tissue. Here, the focus of the study is possible use of mucus released by Phyllocaulis boraceiensis - a compound inducing cell proliferation and enhancing collagen synthesis in dermal fibroblasts and inducing proliferation human endothelial cell cultures. Fibroblasts treated with P. boraceiensis mucus at concentrations below 0.012 μg/μl developed high rates of proliferation, as evaluated using MTT assay; the proliferative effect was dose-dependent. Production and secretion of extracellular matrix components and collagen type I fibres were enhanced after 24 h of treatment, revealing a hormesis effect, biphasic dose response - low dose for proliferation yet toxic at high dose. No significant change in proliferation was observed in treated endothelial cells and production of lipid polyunsaturated free radicals was low in both cell types. Treatment with P. boraceiensis mucus produced pronounced changes in fibroblast cell number and morphology, and in quantities of well-ordered collagen deposition. These results support the premise that Phyllocaulis boraceiensis mucus demonstrates proliferative properties in cells involved in the healing process.

The healing process and wound closure were efficient after the daily application of 0.012 μg/μL P boraceiensis mucus.

Influenza viruses cause worldwide outbreaks and pandemics in humans and animals every year with considerable morbidity and mortality. The molecular diversity of secondary metabolites extracted from mollusks is a good alternative for the discovery of novel bioactive compounds with unique structures and diverse biological activities. Phyllocaulis boraceiensis is a hermaphroditic slug that exudes mucus, in which was detected hydroxy polyunsaturated fatty acids that exhibited potent antiviral activity against measles virus. The objective of this study was to evaluate this property against Influenza viruses. Cell viability and toxicity of the mucus were evaluated on Madin-Darby canine kidney (MDCK) cells by MTT assay. Antiviral activity from mucus against influenza viruses was carried out by determination of the virus infection dose and by immunofluorescence assays. The crude mucus and its fractions exhibited low cytotoxicity on MDCK cells. A significant inhibition of viral replication, reduced by the order of eight times, was observed in influenza-induced cytopathic effect. In immunofluorescence assay was observed a decrease of more than 80% of the viral load on infected MDCK cell treated with mucus and its fractions. The viral glycoproteins hemagglutinin and neuraminidase located on the surface of the virus are crucial for the replications and infectivity of the influenza virus. Some authors demonstrated that lipids, such as, polyunsaturated fatty acids exhibited multiple roles in antiviral innate and adaptive responses, control of inflammation, and in the development of antiviral therapeutics. As corroborated by other studies, hydroxy polyunsaturated fatty acids interfered with the binding of influenza virus on host cell receptor and reduced viral titers. The results obtained indicated that polyunsaturated fatty acids from P. boraceiensis crude mucus and fractions 39 exerted antiviral activity against influenza virus.

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Phyllocaulis boraceiensis mucus is known to be a compound capable of inducing cell proliferation and enhancing the wound healing process. The process of angiogenesis is a chain of mechanisms responsible for the formation of new vessels, which are are involved in cell proliferation, and factors that will act in the healing process. Our aim was to demonstrate that the angiogenesis process is enhanced in cultures of endothelial cells and fibroblasts treated with P. boraceiensis mucus. Experiments were carried out with 10 5 cellsÁmL À1 of endothelial cells and fibroblasts treated with P. boraceiensis mucus in concentrations that have significant effects in proliferation assays, i.e. 0.012 lgÁlL À1 and 0.18 lgÁlL À1 , both of which cause extreme responses. Aliquots of 100 lL of cell suspensions were incubated for 1 h at 4°C with 1 lL of antibodies specific for the cell markers vascular endothelial growth factor receptor 1 and cluster of differentiation 34, and negative isotype controls. Reading and expression analysis of cell markers was performed on a FACSCalibur flow cytometer. Expression levels of vascular endothelial growth factor receptor 1 and cluster of differentiation 34 expression were significantly increased in endothelial cells cultivated with 0.012 lgÁlL À1 P. boraceiensis mucus, suggesting that this compound is capable of enhancing angiogenesis.