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      Propolis Extract for Onychomycosis Topical Treatment: From Bench to Clinic

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          Abstract

          Onychomycosis is a chronic fungal infection of nails, commonly caused by dermatophyte fungi, primarily species of Trichophyton. Because of the limited drug arsenal available to treat general fungal infections and the frequent failure of onychomycosis treatment, the search for new therapeutic sources is essential, and topical treatment with natural products for onychomycosis has been encouraged. Propolis, an adhesive resinous compound produced by honeybees ( Apis mellifera), has shown multiple biological properties including significant antifungal and anti-biofilm activities in vitro. In spite of promising in vitro results, in vivo results have not been reported so far. This study assessed an ethanol propolis extract (PE) as a topical therapeutic option for onychomycosis, including its characterization in vitro and its applicability as a treatment for onychomycosis (from bench to clinic). The in vitro evaluation included analysis of the cytotoxicity and the antifungal activity against the planktonic cells and biofilm formed by Trichophyton spp. We also evaluated the capacity of PE to penetrate human nails. Patients with onychomycosis received topical PE treatments, with a 6-month follow-up period. The results of the in vitro assays showed that PE was non-toxic to the cell lines tested, and efficient against both the planktonic cells and the biofilm formed by Trichophyton spp. The results also showed that PE is able to penetrate the human nail. The results for PE applied topically to treat onychomycosis were promising, with complete mycological and clinical cure of onychomycosis in 56.25% of the patients. PE is an inexpensive commercially available option, easy to obtain and monitor. Our results indicated that PE is a promising natural compound for onychomycosis treatment, due to its ability to penetrate the nail without cytotoxicity, and its good antifungal performance against species such as Trichophyton spp. that are resistant to conventional antifungals, both in vitro and in patients.

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          Most cited references68

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          Antimicrobial activity of flavonoids.

          Flavonoids are ubiquitous in photosynthesising cells and are commonly found in fruit, vegetables, nuts, seeds, stems, flowers, tea, wine, propolis and honey. For centuries, preparations containing these compounds as the principal physiologically active constituents have been used to treat human diseases. Increasingly, this class of natural products is becoming the subject of anti-infective research, and many groups have isolated and identified the structures of flavonoids possessing antifungal, antiviral and antibacterial activity. Moreover, several groups have demonstrated synergy between active flavonoids as well as between flavonoids and existing chemotherapeutics. Reports of activity in the field of antibacterial flavonoid research are widely conflicting, probably owing to inter- and intra-assay variation in susceptibility testing. However, several high-quality investigations have examined the relationship between flavonoid structure and antibacterial activity and these are in close agreement. In addition, numerous research groups have sought to elucidate the antibacterial mechanisms of action of selected flavonoids. The activity of quercetin, for example, has been at least partially attributed to inhibition of DNA gyrase. It has also been proposed that sophoraflavone G and (-)-epigallocatechin gallate inhibit cytoplasmic membrane function, and that licochalcones A and C inhibit energy metabolism. Other flavonoids whose mechanisms of action have been investigated include robinetin, myricetin, apigenin, rutin, galangin, 2,4,2'-trihydroxy-5'-methylchalcone and lonchocarpol A. These compounds represent novel leads, and future studies may allow the development of a pharmacologically acceptable antimicrobial agent or class of agents.
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            Phenolic compounds from Brazilian propolis with pharmacological activities.

            Four compounds were isolated from Brazilian propolis. They are identified as: (1) 3-prenyl-4-hydroxycinnamic acid (PHCA), (2) 2,2-dimethyl-6-carboxyethenyl-2H-1-benzopyrane (DCBEN), (3) 3,5-diprenyl-4-hydroxycinnamic acid (DHCA), and (4) 2,2-dimethyl-6-carboxyethenyl-8-prenyl-2H-1-benzopyran (DPB). The structures of the compounds were determined by MS and NMR techniques. All compounds were assayed against Trypanosoma cruzi and the bacteria Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus faecalis. Compounds (1) to (4) were active against T. cruzi. Except (1), all compounds presented activity against the bacteria tested. When compounds (1)-(3) were tested in the guinea pig isolated trachea, all induced a relaxant effect similar to propolis extract.
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              Potential role of propolis in wound healing: Biological properties and therapeutic activities.

              Propolis is a resinous mixture that honey bees collect from the tree buds, sap flows, or other botanical sources. The chemical composition of propolis varies and depends on the geographical area, time of collection, seasonality, illumination, altitude, and food availability during propolis exploitation. The goal of this review is to discuss important concepts including mechanisms of action and therapeutic activities of propolis. The PubMed, ScienceDirect, and Cochrane Library databases were searched for the literature published from January the 1st 2000 to October the 1st 2017. Sixteen animals and three clinical studies were included. A quantitative and qualitative review was performed on the clinical trials and the animal studies were comprehensively overviewed. In this study, the clinical trials have been combined and the results were provided as meta-analysis. Propolis is a non-toxic natural product; however some cases of allergy and contact dermatitis to this compound have been described mainly among beekeepers. An important factor in impaired wound healing is biofilm formation; propolis as an anti-microbial agent can reduce biofilm generation and result in accelerated healing processes. Most of the in vivo studies on various wound models suggested the beneficial roles of propolis on experimental wound healing and this has also been approved in the clinical trial studies. However, there is a lack of information concerning, dose, side effects and clinical effectiveness of propolis on wounds. As the effectiveness of propolis between different products is variable, more characterizations should be done and future investigations comparing different propolis based products and characterization of their specific roles on different models of wounds are highly appreciated.
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                Author and article information

                Contributors
                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                25 April 2018
                2018
                : 9
                : 779
                Affiliations
                [1] 1Laboratório de Micologia Médica, Departamento de Análises Clínicas e Biomedicina, Universidade Estadual de Maringá , Maringá, Brazil
                [2] 2Programa de Pós-Graduação em Física, Departamento de Física, Universidade Estadual de Maringá , Maringá, Brazil
                [3] 3Microbiology, Parasitology and Pathology Post-Graduation Program, Department of Pathology, Federal University of Paraná , Curitiba, Brazil
                [4] 4Laboratory of Research and Development of Drug Delivery Systems, Department of Pharmacy, Universidade Estadual de Maringá , Maringá, Brazil
                Author notes

                Edited by: Edvaldo Antonio Ribeiro Rosa, Pontifícia Universidade Católica do Paraná, Brazil

                Reviewed by: Taissa Vila, University of Maryland, Baltimore, United States; Priya Uppuluri, University of California, Los Angeles, United States

                *Correspondence: Terezinha I. E. Svidzinski, tiesvidzinski@ 123456uem.br

                This article was submitted to Antimicrobials, Resistance and Chemotherapy, a section of the journal Frontiers in Microbiology

                Article
                10.3389/fmicb.2018.00779
                5996904
                29922236
                42cdbba4-6c53-4bc7-aee4-440034534d1b
                Copyright © 2018 Veiga, Gadelha, da Silva, Costa, Kischkel, de Castro-Hoshino, Sato, Baesso, Voidaleski, Vasconcellos-Pontello, Vicente, Bruschi, Negri and Svidzinski.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 30 January 2018
                : 05 April 2018
                Page count
                Figures: 7, Tables: 4, Equations: 0, References: 75, Pages: 13, Words: 0
                Categories
                Microbiology
                Original Research

                Microbiology & Virology
                dermatophytosis,natural products,propolis,antifungal activity,ex vivo nail model,photoacoustic spectroscopy,permeation property

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