Biological Activity and Chemical Composition of Propolis Extracts with Potential Use in Vulvovaginal Candidiasis Management.

Environmental sustainability is an increasing challenge in the pharmaceutical field, leading to the search for eco-friendly active ingredients. Among natural ingredients, propolis arises as an excellent alternative, being a complex substance with pharmacological properties. This work aims to explore the potential of propolis as a new pharmaceutical ingredient for the replacement of conventional vulvovaginal antifungals. Propolis extracts were obtained by Ultrasound-Assisted Extraction using different solvents (water, water/ethanol (50:50, v/v), and ethanol). Afterwards, the extracts were characterized regarding total phenolic content (TPC), antioxidant/antiradical activities, radical scavenging capacity, antifungal activity against strains of Candida species, and viability effect on two female genital cell lines. The aqueous extract achieved the best TPC result as well as the highest antioxidant/antiradical activities and ability to capture reactive oxygen species. A total of 38 phenolic compounds were identified and quantified by HPLC, among which ferulic acid, phloridzin and myricetin predominated. Regarding the anti-Candida spp. activity, the aqueous and the hydroalcoholic extracts achieved the best outcomes (with MIC values ranging between 128 and 512 µg/mL). The cell viability assays confirmed that the aqueous extract presented mild selectivity, while the hydroalcoholic and alcoholic extracts showed higher toxicities. These results attest that propolis has a deep potential for vulvovaginal candidiasis management, supporting its economic valorization.

Assessment of the Antioxidant Capacity of Commercial Coffee Using Conventional Optical and Chromatographic Methods and an Innovative Electrochemical DNA-Based Biosensor.

As one of the most popular beverages in the world, coffee is a rich source of non-enzymatic bioactive compounds with antioxidant capacity. In this study, twelve commercial coffee beverages found in local Portuguese markets were assessed to determine their total phenolic and flavonoid contents, as well as their antioxidant capacity, by conventional optical procedures, namely, ferric reducing antioxidant power and DPPH-radical scavenging assay, and non-conventional procedures such as a homemade DNA-based biosensor against two reactive radicals: HO• and H2O2. The innovative DNA-based biosensor comprised an adenine-rich oligonucleotide adsorbed onto a carbon paste electrode. This method detects the different peak intensities generated by square-wave voltammetry based on the partial damage to the adenine layer adsorbed on the electrode surface by the free radicals in the presence/absence of antioxidants. The DNA-based biosensor against H2O2 presented a higher DNA layer protection compared with HO• in the presence of the reference gallic acid. Additionally, the phenolic profiles of the twelve coffee samples were assessed by HPLC-DAD, and the main contributors to the exhibited antioxidant capacity properties were caffeine, and chlorogenic, protocatechuic, neochlorogenic and gallic acids. The DNA-based sensor used provides reliable and fast measurements of antioxidant capacity, and is also cheap and easy to construct.

3D-printed microgels supplemented with dentin matrix molecules as a novel biomaterial for direct pulp capping.

OBJECTIVES: To develop a 3D-printed, microparticulate hydrogel supplemented with dentin matrix molecules (DMM) as a novel regenerative strategy for dental pulp capping. MATERIALS AND METHODS: Gelatin methacryloyl microgels (7% w/v) mixed with varying concentrations of DMM were printed using a digital light projection 3D printer and lyophilized for 2 days. The release profile of the DMM-loaded microgels was measured using a bicinchoninic acid assay. Next, dental pulp exposure defects were created in maxillary first molars of Wistar rats. The exposures were randomly capped with (1) inert material - negative control, (2) microgels, (3) microgels + DMM 500 µg/ml, (4) microgels + DMM 1000 µg/ml, (5) microgels + platelet-derived growth factor (PDGF 10 ng/ml), or (6) MTA (n = 15/group). After 4 weeks, animals were euthanized, and treated molars were harvested and then processed to evaluate hard tissue deposition, pulp tissue organization, and blood vessel density. RESULTS: All the specimens from groups treated with microgel + 500 µg/ml, microgel + 1000 µg/ml, microgel + PDGF, and MTA showed the formation of organized pulp tissue, tertiary dentin, newly formed tubular and atubular dentin, and new blood vessel formation. Dentin bridge formation was greater and pulp necrosis was less in the microgel + DMM groups compared to MTA. CONCLUSIONS: The 3D-printed photocurable microgels doped with DMM exhibited favorable cellular and inflammatory pulp responses, and significantly more tertiary dentin deposition. CLINICAL RELEVANCE: 3D-printed microgel with DMM is a promising biomaterial for dentin and dental pulp regeneration in pulp capping procedures.

New Perspectives on the Sustainable Employment of Chestnut Shells as Active Ingredient against Oral Mucositis: A First Screening.

Oral mucositis (OM), a common side effect of oncological treatment, is an oral mucosal disorder characterized by painful ulcerations and increased risk of infection. The use of natural antioxidants to suppress the redox imbalance responsible for the OM condition has emerged as an interesting approach to prevent/treat OM. This study aims to explore the chestnut (Castana sativa) shells as potential active ingredient against OM. Therefore, chestnut shells were extracted at different temperatures (110-180 °C) by Subcritical Water Extraction (SWE), aiming to recover antioxidants. The extracts were also evaluated against microorganisms present in the oral cavity as well as on human oral cell lines (TR146 and HSC3). The highest phenolic content was obtained with the extraction temperature of 110 °C, exhibiting the best antioxidant/antiradical activities and scavenging efficiencies against HOCl (IC50 = 4.47 µg/mL) and ROO• (0.73 µmol TE/mg DW). High concentrations of phenolic acids (e.g., gallic and protocatechuic acids) and flavanoids (catechin, epicatechin and rutin) characterized the phenolic profile. The antimicrobial activity against several oral microorganisms present in the oral cavity during OM, such as Streptococcus, Staphylococcus, Enterococcus, and Escherichia, was demonstrated. Finally, the effects on HSC3 and TR146 cell lines revealed that the extract prepared at 110 °C had the lowest IC50 (1325.03 and 468.15 µg/mL, respectively). This study highlights the potential effects of chestnut shells on OM.

Valorization of Phenolic and Carotenoid Compounds of Sechium edule (Jacq. Swartz) Leaves: Comparison between Conventional, Ultrasound- and Microwave-Assisted Extraction Approaches.

Chayote leaves are known for culinary and traditional medicine applications. This work intended to recover carotenoids and phenolic compounds from chayote leaves using the ultrasound-assisted extraction (UAE). A Box-Behnken design was employed to investigate the impact of extraction time, temperature, and ultrasonic power on the recovery of total carotenoids, total phenolic compounds, and antioxidant activities. For comparative purposes, chayote leaf extracts were prepared by maceration (ME) and microwave-assisted extraction (MAE), using the same time and temperature conditions optimized by UAE. Extraction at 50 °C and 170 Watts for 30 min provided the optimal UAE conditions. UAE showed better extraction efficacy than ME and MAE. The HPLC analysis of the extracts showed that the xanthophyll class was the main class of carotenoids, which constituted 42-85% of the total carotenoid content, followed by ß-carotene and tocopherol. Moreover, 26 compounds, classified as phenolic acids, flavonols, flavonoids and other polar compounds, were identified in the chayote leaf extracts. Flavonols accounted for 55% of the total compounds quantified (the major compound was myricetin) and phenolic acids represented around 35%, mostly represented by ferulic acid, chlorogenic acid and (+)-catechin. This study revealed the potential of UAE as an effective green extraction technique to recover bioactive compounds from chayote leaves, for food, and for pharmaceutical and cosmetic applications.

Increasing the added value of vine-canes as a sustainable source of phenolic compounds: A review.

Grapes represent one of the most produced fruit crops around the world leading to the generation of large amounts of vine-canes as a side product, with no current economically profitable application. However, vine-canes have been demonstrated to be natural sources of phenolic compounds with numerous health benefits associated, with several potential applications. Therefore, new ambitious applications focused on their re-use are needed, targeting a sustainable process that simultaneous produces functional products and mitigates the waste generation. This review gives to the readers a complete summary about the state of the art regarding the vine-canes extracts research. Vine-canes phenolic composition is addressed and related to the health benefits exhibited. This review comprises studies from the past two decades reporting the extraction processes to recover vine-cane phenolic compounds, including conventional and environmentally friendly technologies and discussing their advantages and disadvantages. The conditions that favour the extraction process for vine-cane polyphenols for each technique were also deeply explored for the first time, enabling to the reader apply only the best parameters to achieve the highest yields without huge investment in optimizations procedures. Furthermore, a correlation between the bioactive properties of the vine-cane extracts and their applications in multiple fields is also critically presented.

Vine-Canes as a Source of Value-Added Compounds for Cosmetic Formulations.

The majority of works about vine-canes are focused on the evaluation of their chemical composition and antioxidant potential. To the best of our knowledge, the possible applications of produced extracts in cosmetic formulations have never been explored. The aim of the present study was to evaluate the antioxidant properties of vine-canes subcritical water extracts for use as active ingredients in the cosmetic industry. For that, the phenolic content and antioxidant activity of six vine-cane varieties, namely Alvarinho and Loureiro from the Minho region and Touriga Nacional and Tinta Roriz (TR) from both the Douro and Dão regions, were evaluated through spectrophotometric and chromatographic methods. All extracts presented similar antioxidant activity and the highest phenolic content was reported for TR variety from the Douro region (33.7 ± 1.9 mg GAE/g dw). The capacity of vine-cane extracts to capture reactive oxygen species superoxide (O2-) was also studied, with the highest IC50 value being obtained for Loureiro variety (56.68 ± 2.60 µg/mL). Furthermore, no adverse effects on HaCaT and HFF-1 dermal cell lines in concentrations below 100 and 1000 µg/mL, respectively, were determined. Finally, Loureiro vine-cane extract was incorporated into a topical formulation, and physical and microbiological properties were within expected values, demonstrating that vine-canes extracts can be successfully incorporated in cosmetic products.

Vine-Canes Valorisation: Ultrasound-Assisted Extraction from Lab to Pilot Scale.

Wine production generates large amounts of vine-canes, a devalued by-product that could be used for the recovery of bioactive compounds. In this work, two vine-canes varieties, namely Touriga Nacional (TN) and Tinta Roriz (TR), were submitted to different ultrasound-assisted extraction (UAE) conditions. The highest phenolic and flavonoid content was observed for TR extract obtained at lab-scale without an ice bath and pilot-scale after 60 min of extraction (32.6 ± 2.1 and 26.0 ± 1.5 mg gallic acid equivalent/g dry weight (dw) and 9.5 ± 0.6 and 8.3 ± 0.8 mg epicatechin equivalents/g dw, respectively). Further, all extracts demonstrated a high antioxidant activity to scavenge DPPH free radicals with the best value reached by TR at the lab-scale without an ice bath after 30 min and pilot-scale extraction after 60 min (34.2 ± 2.4 and 33.4 ± 2.1 mg trolox equivalents/g dw, respectively). Extracts phenolic composition were also evaluated by HPLC, demonstrating that resveratrol, myricetin and catechin were the main compounds. According to our knowledge, this is the first time that a pilot scale of UAE of phenolic compounds from vine-canes was performed. This paper represents an important step to the use of UAE as an industrial process to recover bioactive compounds.

Antioxidant, photoprotective and inhibitory activity of tyrosinase in extracts of Dalbergia ecastaphyllum.

Dalbergia ecastaphyllum is a native Brazil plant with importance for beekeeping, and widely used in folk medicine. For the first time, the extracts of this plant were assessed for the presence of hydrophilic and lipophilic antioxidants, as well as inhibition of tyrosinase, free radicals scavenging and sunscreen protection. The antioxidant activity was evaluated by free radical scavenging (DPPH) and ß-carotene bleaching assay. The tyrosinase inhibitory activity was evaluated and calculated the EC50. The photoprotective activity was measured using different concentrations of D. ecastaphyllum extracts. The Sun Protection Factor (SPF) of the samples was higher than 6, and the sample from Ilhéus showed the most pronounced photoprotective effect. Sample from Canavieiras presented the highest antioxidant activity by free radical scavenging DPPH and ß-carotene bleaching method, with 92.41% and 48.34%, respectively. All samples inhibited the tyrosinase, especially the sample from Prado that was most effective (124.62 µg.mL-1). Significant negative correlation was found between flavonoid contents and inhibition of tyrosinase. The overall results provide relevant information about the Dalbergia ecastaphyllum species, indicating as potential material to cosmetic and pharmaceutical industry.

Potential of Portuguese vine shoot wastes as natural resources of bioactive compounds.

Since annually a high amount of wastes is produced in vine pruning, the aim of this study was to evaluate the potential of vine shoots from two Portuguese grape varieties (Touriga Nacional - TN and Tinta Roriz - TR) to be used as a natural source of phenolic compounds. To reach this goal, three techniques were explored, namely microwave-assisted extraction (MAE), subcritical water extraction (SWE) and conventional extraction (CE). The phenolic composition of the extracts, antioxidant and biological activities were evaluated by spectrophotometry and chromatography. MAE and SWE produced the highest concentrated extracts. TR vine shoot variety had the highest antioxidant activity and total phenolic (32.1±0.9mggallicacidequivalents/g dry sample), as well as flavonoid content (18.7±1.2mgepicatechinequivalents/g dry sample). For the first time, the biological activity of the vine shoot extracts was tested. Results demonstrated that all of them had antimicrobial potential against different bacteria and yeasts, and the ability of inhibiting α-amylase and acetylcholinesterase enzymes, with MAE TR extracts being the most efficient. HPLC analysis enabled the identification of different phenolic compounds, with gallic acid, catechin, myricetin and kaempferol-3-O-rutinoside being the main contributors to the phenolic composition. Portuguese vine shoot wastes could serve as easily accessible source of natural antioxidants for the food or pharmaceutical industries.

An Insight on Salting-out Assisted Liquid-Liquid Extraction for Phytoanalysis.

INTRODUCTION: Salting-out assisted liquid-liquid extraction (SALLE) is a technique that, although simple and not requiring any complex equipment, is very powerful and versatile. It has obtained growing interest in bioanalysis particularly when combined with chromatographic techniques. OBJECTIVES: Herein, fennel seeds (Foeniculum vulgare Mill.) were used as a case-study to show the application of SALLE in phytochemical analysis. MATERIAL AND METHODS: SALLE combined with HPLC-UV-MS/MS and GC-MS. RESULTS: By HPLC-UV-MS/MS analysis of the organic extract it was possible to identify various phenolic compounds, including quercetin derivatives, caffeic acid, p-coumaric acid and chlorogenic acid. The main compounds identified by GC-MS were estragole, fenchone, anisaldehyde, anethole, benzaldehyde, camphor and apiole. CONCLUSION: HPLC and GC analysis of the extracts showed that it is possible to isolate, in only one step, a wide range of compounds with distinct properties, allowing a detailed phytochemical analysis. Copyright © 2017 John Wiley & Sons, Ltd.

A novel application of microwave-assisted extraction of polyphenols from brewer's spent grain with HPLC-DAD-MS analysis.

This paper reports a novel application of microwave-assisted extraction (MAE) of polyphenols from brewer's spent grains (BSG). A 2(4) orthogonal composite design was used to obtain the optimal conditions of MAE. The influence of the MAE operational parameters (extraction time, temperature, solvent volume and stirring speed) on the extraction yield of ferulic acid was investigated through response surface methodology. The results showed that the optimal conditions were 15 min extraction time, 100 °C extraction temperature, 20 mL of solvent, and maximum stirring speed. Under these conditions, the yield of ferulic acid was 1.31 ± 0.04% (w/w), which was fivefold higher than that obtained with conventional solid-liquid extraction techniques. The developed new extraction method considerably reduces extraction time, energy and solvent consumption, while generating fewer wastes. HPLC-DAD-MS analysis indicated that other hydroxycinnamic acids and several ferulic acid dehydrodimers, as well as one dehydrotrimer were also present, confirming that BSG is a valuable source of antioxidant compounds.

Novel application of square-wave adsorptive-stripping voltammetry for the determination of xanthohumol in spent hops.

This paper reports the development of a novel electrochemical assay for xanthohumol (XN) by square-wave adsorptive-stripping voltammetry (SWAdSV) with a hanging mercury drop electrode. The method showed good repeatability (CV < 2%) and linearity (between 10 and 250 µg L(-1)), as well as suitable limits of detection (2.6 µg L(-1)) and quantification (8.8 µg L(-1)). The method was applied for the quantification of this compound in spent hops, and the results obtained were compared with the HPLC-UV method. XN contents determined by the SWAdSV method were 16 ± 1 and 100 ± 4 µg L(-1) for aqueous and methanolic extracts, respectively. The developed new methodology considerably reduces the analysis time, approximately from 25 min (HPLC-UV method) to 7 min, enabling a high sample throughput. In addition, the detection and quantification limits were approximately 5-fold lower than those obtained with the chromatographic method.