Preventing microbial infections with natural phenolic compounds

The struggle between humans and pathogens has taken and is continuing to take countless lives every year. As the misusage of conventional antibiotics increases, the complexity associated with the resistance mechanisms of pathogens has been evolving into gradually more clever mechanisms, diminishing the effectiveness of antibiotics. Hence, there is a growing interest in discovering novel and reliable therapeutics able to struggle with the infection, circumvent the resistance and defend the natural microbiome. In this regard, nature-derived phenolic compounds are gaining considerable attention due to their potential safety and therapeutic effect. Phenolic compounds comprise numerous and widely distributed groups with different biological activities attributed mainly to their structure. Investigations have revealed that phenolic compounds from natural sources exhibit potent antimicrobial activity against various clinically relevant pathogens associated with microbial infection and sensitize multi-drug resistance strains to bactericidal or bacteriostatic antibiotics. This review outlines the current knowledge about the antimicrobial activity of phenolic compounds from various natural sources, with a particular focus on the structure-activity relationship and mechanisms of actions of each class of natural phenolic compounds, including simple phenols, phenolic acids, coumarin, flavonoids, tannins, stilbenes, lignans, quinones, and curcuminoids.The authors acknowledge the financial support by the Portuguese Foundation for Science and Technology (FCT) through the doctoral grant and junior research contract with the reference number PD/BD/150521/2019 (K.E.) and CEECIND/01026/2018 (J.M.S.), respectively

New endoscopic procedure for bladder wall closure: results from the porcine model

Upper urinary tract urothelial carcinomas are usually managed by radical nephroureterectomy (RNU), often followed by intravesical chemotherapy to minimize recurrence. Open surgery is the gold standard procedure for RNU, but it associates with high morbidity, and it has been increasingly replaced by minimally invasive strategies, such as laparoscopy and endoscopy. Although effective, endoscopic ureteral excision leaves the bladder unsutured, increasing the risk of tumor spillage, and precluding the immediate administration of intravesical chemotherapy. Here we describe a new method to close the bladder wall after ureteral excision, using barbed sutures via the endoscopic access. Our results in 8 female pigs demonstrate that this method is effective to close the bladder wall. The procedure was completed in a median time of 24 min, and no adverse events were registered in the follow-up or at the three-week necropsy. This technique improves a previous approach described by our group because the device is more flexible and allows to tie the knots inside the bladder. Barbed sutures have been used in the clinical practice for other types of surgeries, and therefore this method can further be adapted to human patients with no safety concerns. Its use may allow to administer intravesical chemotherapy, which reduces tumor recurrence and improves patient outcomes.The authors acknowledge Ana Goios for support in manuscript writing and technical editing, and for producing original illustrations for Figure 3. Doctoral grant of CUF Jose de Mello Saude

Tailor made degradable ureteral stents from natural origin polysaccharides

A urinary stent is defined as a thin tube, which is inserted in the ureter to prevent or treat the obstruction of urine flow from the kidney. Silicone, latex, polyvinylchloride and polyurethanes are the most widely used materials for the preparation of stents. Nonetheless, severe clinical complications may result from the use of these materials such as fracture, encrustation and infection. In some of the cases, the ureteral stents are temporary and it is often required a second surgery to remove the stent. The main complications with ureteral stents are dislocation, infection, and blockage by encrustation [1]. Recently, a tendency has been noticed favouring less invasive approaches (e.g. pharmacological or catheterization) in treating patients who exhibit symptoms or signs of urinary retention [2]. Currently, nearly 100% of the people who have an urological stent are likely to develop a bacterial infection within 30 days, which increases morbidity threefold [1]. Different types of temporary and permanent stents have been introduced into urological practice to relieve obstructions [3]. Particular attention should be devoted to polymers as they represent a highly versatile class of materials. Despite the fact that silicon continues to be the gold standard material for urological stents, there have been fast developments in manufacturing processes, as well as the introduction of new biodegradable materials in order to overcome the drawbacks of the available products. Polyurethane continues to be the most widely used material for polymeric stents; however it frequently promotes biofilm formation and bacterial adhesion leading to severe infections [2]. The concerns regarding existing stents are the motivation to design new biodegradable urological stent systems based on natural polymers, specifically polysaccharides, which present inherent biocompatibility, anti-bacterial properties and that can be tailor-made into a custom suitable stent for a particular patient

Bioresorbable ureteral stents from natural origin polymers

In this work, stents were produced from natural originpolysaccharides. Alginate, gellan gum, and a blend ofthese with gelatin were used to produce hollow tube (stents)following a combination of templated gelation and criticalpoint carbon dioxide drying. Morphological analysis of thesurface of the stents was carried out by scanning electronmicroscopy. Indwelling time, encrustation, and stability ofthe stents in artificial urine solution was carried out up to 60days of immersion. In vitro studies carried out with simulatedurine demonstrated that the tubes present a high fluid uptakeability, about 1000%. Despite this, the materials are able tomaintain their shape and do not present an extensive swellingbehavior. The bioresorption profile was observed to behighly dependent on the composition of the stent and it canbe tuned. Complete dissolution of the materials may occurbetween 14 and 60 days. Additionally, no encrustation wasobserved within the tested timeframe. The ability to resistbacterial adherence was evaluated with Gram-positive Staphylococcus aureus and two Gram-negatives Escherichia coliDH5 alpha and Klebsiella oxytoca. For K. oxytoca, no differenceswere observed in comparison with a commercialstent (BiosoftVR duo, Porges), although, for S. aureus alltested compositions had a higher inhibition of bacterialadhesion compared to the commercial stents. In case ofE. coli, the addition of gelatin to the formulations reducedthe bacterial adhesion in a highly significant manner comparedto the commercial stents. The stents produced by thedeveloped technology fulfill the requirements for ureteralstents and will contribute in the development of biocompatible and bioresorbable urinary stents.Contract grant sponsor: the European Union Seventh Framework Programme (FP7/2007-2013); contract grant number: REGPOT-CT2012-316331POLARIS Contract grant sponsor: Novel smart and biomimetic materials for innovative regenerative medicine approaches”; contract grant number: RL1 – ABMR – NORTE-01-0124-FEDER-000016; North Portugal Regional Operational Programme (ON.2 – O Novo Norte), National Strategic Reference Framework (NSRF), European Regional Development Fund (ERDF) Contract grant sponsor: FCT post-doc; contract grant numbers: SFRH/BPD/39333/2007, SFRH/BPD/90533/20

Water and carbon dioxide: green solvents for the extraction of collagen/gelatin from marine sponges

"Publication Date (Web): December 23, 2014"Marine sponges are extremely rich in natural products and are considered a promising biological resource. The major objective of this work is to couple a green extraction process with a natural origin raw material to obtain sponge origin collagen/gelatin for biomedical applications. Marine sponge collagen has unique physicochemical properties, but its application is hindered by the lack of availability due to inefficient extraction methodologies. Traditional extraction methods are time consuming as they involve several operating steps and large amounts of solvents. In this work, we propose a new extraction methodology under mild operating conditions in which water is acidified with carbon dioxide (CO2) to promote the extraction of collagen/gelatin from different marine sponge species. An extraction yield of approximately 50% of collagen/gelatin was achieved. The results of Fourier transformed infrared spectroscopy (FTIR), circular dichroism (CD), and differential scanning calorimetry (DSC) spectra suggest a mixture of collagen/gelatin with high purity, and the analysis of the amino acid composition has shown similarities with collagen from other marine sources. Additionally, in vitro cytotoxicity studies did not demonstrate any toxicity effects for three of the extracts.The authors are grateful for financial support of FCT through Grant EXP/QEQ:EPS/0745/2012, SWIMS (Subcritical Water Isolation of compounds from Marine Sponges). The funding from the European Union Seventh Framework Programme (FP7/2007-2013) under grant agreement numbers REGPOT-CT2012-316331-POLARIS and KBBE-2010-266033 (project SPECIAL), as well as from ERDF under the project "Novel smart and biomimetic materials for innovative regenerative medicine approaches" RLI-ABMR-NORTE-01-0124-FEDER-000016), cofinanced by North Portugal Regional Operational Programme (ON.2,O Novo Norte), under the National Strategic Reference Framework (NSRF) are also gratefully ackowledged. The authors are also truly thankfull to Prof. Micha flan (Tel Aviv University, Israel), Dr. Ronald Osinga (Porifarma, The Netherlands), Dr. Antonio Sara and Dr. Martina Milanese (Studio Associato GAIA, Italy), and Dr. Joana Xavier (University of Azores) for the kind offer of marine sponges samples

Surface modification of silica-based marine sponge bioceramics induce hydroxyapatite formation

Marine biomaterials are a new emerging area of research with significant applications. Recently, researchers are dedicating considerable attention to marine-sponge biomaterials for various applications. We have focused on the potential of biosilica from Petrosia ficidormis for novel biomedical/industrial applications. A bioceramic structure from this sponge was obtained after calcination at 750ºC for 6 hours in a furnace. The morphological characteristics of the 3D architecture were evaluated by scanning electron microscopy (SEM) and micro-computed tomography revealing a highly porous and interconnected structure. The skeleton of Petrosia ficidormis is a siliceous matrix composed of SiO2, which does not present inherent bioactivity. Induction of bioactivity was attained by subjecting the bioceramics structure to an alkaline treatment (KOH 2M) and acidic treatment (HCl 2M) for 1 and 3 hours. In vitro bioactivity of the bioceramics structure was evaluated in simulated body fluid (SBF), after 7 and 14 days. Observation of the structures by SEM, coupled with spectroscopic elemental analysis (EDS), has shown that the surface morphology presented a calcium-phosphate CaP coating, similar to hydroxyapatite (HA). The determination of the Ca/P ratio, together with the evaluation of the characteristic peaks of HA by infra-red spectroscopy and X-ray diffraction, have proven the existence of HA. In vitro biological performance of the structures was evaluated using an osteoblast cell line andthe acidic treatment has shown to be the most effective treatment. Cells were seeded on the bioceramics structures and their morphology, viability and growth was evaluated by SEM, MTS assay and DNA quantification, respectively, demonstrating that cells are able to grow and colonize the bioceramic structures.Alexandre Barros is grateful for financial support of FCT through Grant EXP/QEQ-EPS/0745/2012, SWIMS - Subcritical Water Isolation of compounds from Marine Sponges. The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007-2013) under Grant REGPOT-CT2012-316331-POLARIS and under Grant no KBBE-2010-266033 (project SPECIAL). Funding from the project "Novel smart and biomimetic materials for innovative regenerative medicine approaches" RL1-ABMR-NORTE-01-0124-FEDER-000016) cofinanced by North Portugal Regional Operational Programme (ON.2 - O Novo Norte), under the National Strategic Reference Framework (NSRF) is also acknowledged

Preparation of macroporous alginate-based aerogels for biomedical applications

Aerogels are a special class of ultra-light porous materials with growing interest in biomedical applications due to their open pore structure and high surface area. However, they usually lack macroporosity, while mesoporosity is typically high. In this work, carbon dioxide induced gelation followed by expansion of the dissolved CO2 was performed to produce hybrid calcium-crosslinked alginate-starch hydrogels with dual meso- and macroporosity. The hydrogels were subjected to solvent exchange and supercritical drying to obtain aerogels. Significant increase in macroporosity from 2 to 25 % was achieved by increasing expansion rate from 0.1 to 30 bar/min with retaining mesoporosity (BET surface and BJH pore volume in the range 183 â 544 m2/g and 2.0 â 6.8 cm3/g, respectively). In vitro bioactivity studies showed that the alginate-starch aerogels are bioactive, i.e. they form hydroxyapatite crystals when immersed in a simulated body fluid solution. Bioactivity is attributed to the presence of calcium in the matrix. The assessment of the biological performance showed that the aerogels do not present a cytotoxic effect and the cells are able to colonize and grow on their surface. Results presented in this work provide a good indication of the potential of the alginate-starch aerogels in biomedical applications, particularly for bone regeneration. aerogels, alginate, starch, tissue engineering, supercritical fluids, CO2 induced gelation. (undefined

Directional field-dependence of magnetoimpedance effect on integrated YIG/Pt-stripline system

We investigated the magnetization dynamics through the magnetoimpedance effect in an integrated YIG/Pt-stripline system in the frequency range of 0.5 up to 2.0 GHz. Specifically, we explore the dependence of the dynamic magnetic behavior on the field orientation by analyzing beyond the traditional longitudinal magnetoimpedance effect of the transverse and perpendicular setups. We disclose here the strong dependence of the effective damping parameter on the field orientation, as well as verification of the very-low damping parameter values for the longitudinal and transverse configurations. We find considerable sensitivity results, bringing to light the facilities to integrate ferrimagnetic insulators in current and future technological applications.This research was funded by CNPq grand numbers 304943/2020-7 and 407385/2018-5, Capes grand number 88887.573100/2020-00 and FCT grant number CTTI-31/18-CF(2)

Biodegradable drug-eluting stents: Targeting urothelial tumors of upper urinary tract

INTRODUCTION & OBJECTIVES: Urothelial tumors of upper urinary tract are ranked among the most common types of cancers worldwide. The current standard therapy to prevent recurrence is intravesical Bacillus Calmetteâ Guerin (BCG) immunotherapy, but it presents several disadvantages such as BCG failure and intolerance. Another way is to use chemotherapy, which is generally better tolerated that BCG. In this case, drugs such as epirubicin, doxorubicin, paclitaxel and gemcitabine are used. Nevertheless, intravesical chemotherapy only prevents recurrence in the short-term. These failings can be partially attributed to the short residence time and low bioavailability of the drug within the upper urinary tract and the cancer cells, resulting in a need for frequent drug instillation. To avoid these problems, biodegradable ureteral stents impregnated by supercritical fluid CO2 (SCF) with each of the four anti-cancer drugs were produced. MATERIAL & METHODS: Four formulations with different concentrations of gelatin and alginate and crosslink agent were tested and bismuth was added to confer radiopaque properties to the stent. The preliminary in vivo validation studies in female domestic pigs was conducted at the University of Minho, Braga, after formal approval by the institutionâ s review board and in accordance with its internal ethical protocol for animal experiments. Paclitaxel, epirubicin, doxorubicin and gemcitabine were impregnated in the stents and the release kinetics was measured in artificial urine solution (AUS) for 9 days by UV spectroscopy in a microplate reader. The anti-tumoral effect of the developed stents in transitional cell carcinoma (TCC) and HUVEC primary cells, used as control, was evaluated. RESULTS: The in vivo validation of this second-generation of ureteral stents performed was herein demonstrated. Biodegradable ureteral stents were placed in the ureters of a female pigs, following the normal surgical procedure. The animals remained asymptomatic, with normal urine flow. The in vitro release study in AUS of the stent impregnated showed a higher release in the first 72h for the four anti-cancer drugs impregnated after this time the plateau was achieved and the stent degraded after 9 days. The direct and indirect contact of the anti-cancer biodegradable stents with the TCC and HUVEC cell lines confirm the anti-tumor effect of the stents impregnated with the four anti-cancer drugs, reducing around 75% of the viability of the TCC cell line after 72h and no killing effect in the HUVEC cells. CONCLUSIONS: The use of biodegradable ureteral stent in urology clinical practice not only reduce the stent-related symptoms but also open new treatment therapyâ s, like in urothelial tumors of upper urinary tract. Furthermore, we have demonstrated the clinical validation in vivo pig model. This study has thus shown the killing efficacy of the anti-cancer drug eluting biodegradable stents in vitro for the TCC cell line, with no toxicity observed in the control, non-cancerous cells.The direct and indirect contact of the anti-cancer biodegradable stents with the TCC and HUVEC cell lines confirm the anti-tumor effect of the stents impregnated with the four anti-cancer drugs, reducing around 75% of the viability of the TCC cell line after 72h and no killing effect in the HUVEC cells. This study has thus shown the killing efficacy of the anti-cancer drug eluting biodegradable stents in vitro for the TCC cell line, with no toxicity observed in the control, non-cancerous cells

Targeting urothelial tumors of upper urinary tract with drug-eluting stents impregnated by supercritical fluids

Urothelial tumors of upper urinary tract are ranked among the most common types of cancers worldwide and it has been considered as one of the more expensive to treat due of its long-term propensity of recurrence. The current standard therapy to prevent recurrence is intravesical Bacillus Calmette–Guerin (BCG) immunotherapy, but it presents several disadvantages such as BCG failure and intolerance. Another way is to use chemotherapy, that has been reported to be generally better tolerated that BCG. In this case, drugs such as epirubicin, doxorubicin, paclitaxel and gemcitabine are used. Nevertheless, intravesical chemotherapy only prevents recurrence in the short-term[1], [2]. These failings can be partially attributed to the short residence time and low penetration of the drug within the upper urinary tract and the cancer cells, resulting in a need for frequent drug instillation [3]. To avoid these problems, biodegradable ureteral stents impregnated by supercritical fluid CO2 (SCF) with each of the four anti-cancer drugs were produced (figure 1). Four types of drug-eluting biodegradable stents were studied, impregnated with paclitaxel, epirubicin, doxorubicin and gemcitabine. The release kinetics of the impregnated drugs from the anti-cancer drug-eluting stents was measured in artificial urine solution (AUS) for 9 days. The in vitro drugs release from the impregnated biodegradable ureteral stents was analyzed using a microplate reader. The in vitro release study in AUS showed a higher release in the first 72h for the four anti-cancer drugs impregnated after this time the plateau was achieved and the stent degrades after 9 days. Regarding the amount of impregnated drugs by SCF the gemcitabine showed higher amount (109 μg) and the lower amount was obtained for paclitaxel (67 ng). The diffusion coefficient and the impregnation yield were calculated. The anti-tumoral effect of the developed stents in transitional cell carcinoma (TCC) - T24 cell lines was evaluated. T24 cell line was exposed to graded concentrations (0.01 to 2000 ng/ml) of the four drugs for both 4 and 72 hours to determine the sensitivities to each drug (IC50). Toxicity as a result of both direct and indirect contact of the cell lines with the different material conditions of biodegradable stent were studied. The four anti-cancer drugs showed a concentrationdependent inhibitory effect on the T24 and HUVEC cell lines with IC50’s for paclitaxel of 7.30ng and 501.50ng, respectively. The T24 cell line shows to be more sensitive than HUVEC cell line for all the anti-cancer drugs tested. The direct and indirect contact of the anti-cancer biodegradable stents with the T24 and HUVEC cell lines confirm the anti-tumor effect of the stents impregnated with the four anti-cancer drugs, reducing around 75% of the viability of the T24 cell line after 72h and no killing effect in the HUVEC cells. Finally, this study has shown the killing efficacy of the anti-cancer drug eluting biodegradable stents in vitro for the T24 cell lines, with no toxicity observed in the control, non-cancerous cells.Luso­- American Foundation's Grant for Internships in the University of California, Berkeley, 2015/CON5/CAN8; FCT PhD Grant (SFRH/BD/97203/2013); European Union's Seventh Framework Programme (FP7/2007­2013) under grant agreement n° REGPOT­CT2012­316331­ POLARIS; Project “Novel smart and biomimetic materials for innovative regenerative medicine approaches (Ref.: RL1 ­ ABMR ­ NORTE­01­0124­FEDER­000016)” cofinanced by North Portugal Regional Operational Programme (ON.2 – O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF