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Bacterial cellulose (BC), produced by bacterial fermentation, is a high-purity material. BC can be oxidized (BCOXI), providing aldehyde groups for covalent bonds with drugs. Frutalin (FTL) is a lectin capable of modulating cell proliferation and remodeling, which accelerates wound healing. This study aimed to develop an FTL-incorporated dressing based on BC, and to evaluate its physicochemical properties and biological activity in vitro. An experimental design was employed to maximize FTL loading yield onto the BC and BCOXI, where independent variables were FTL concentration, temperature and immobilization time. BCOXI-FTL 1 (44.96 % ± 1.34) had the highest incorporation yield (IY) at the experimental conditions: 6 h, 5 °C, 20 µg mL-1. The second highest yield was BCOXI-FTL 6 (23.28 % ± 1.43) using 24 h, 5 °C, 100 µg mL-1. Similarly, the same reaction parameters provided higher immobilization yields for native bacterial cellulose: BC-FTL 6 (16.91 % ± 1.05) and BC-FTL 1 (21.71 % ± 1.57). Purified FTL displayed no cytotoxicity to fibroblast cells (<50 µg mL-1 concentration) during 24 h. Furthermore, BCOXI-FTL and BC-FTL were non-cytotoxic during 24 h and stimulated fibroblast migration. BCOXI-FTL demonstrated neutrophil activation in vitro similar to FTL. These promising results indicate that the bacterial cellulose matrices containing FTL at low concentrations, could be used as an innovative biomaterial for developing wound dressings.
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Artocarpus , Bandagens , Celulose , Lectinas de Plantas , Artocarpus/química , Celulose/química , Celulose/farmacologia , Lectinas de Plantas/química , Lectinas de Plantas/farmacologia , Animais , Camundongos , Fibroblastos/efeitos dos fármacos , Cicatrização/efeitos dos fármacosRESUMO
Bacterial cellulose (BC) represents a promising biomaterial, due to its unique and versatile properties. We report, herein, on purposely-designed structural modifications of BC that enhance its application as a wound dressing material. Chemical modification of the functional groups of BC was performed initially to introduce a hydrophobic/oleophilic character to its surface. Specifically, silanization was carried out in an aqueous medium using methyltrimethoxisilane (MTMS) as the silanizing agent, and aerogels were subsequently prepared by freeze-drying. The BC-MTMS aerogel obtained displayed a highly porous (99 %) and lightweight structure with an oil absorption capacity of up to 52 times its dry weight. The XRD pattern indicated that the characteristic crystallographic planes of the native BC were maintained after the silanization process. Thermal analysis showed that the thermal stability of the BC-MTMS aerogel increased, as compared to the pure BC aerogel (pBC). Moreover, the BC-MTMS aerogel was not cytotoxic to fibroblasts and keratinocytes. In the second step of the study, the incorporation of natural oils into the aerogel's matrix was found to endow antimicrobial and/or healing properties to BC-MTMS. Bourbon geranium (Pelargonium X ssp.) essential oil (GEO) was the only oil that exhibited antimicrobial activity against the tested microorganisms, whereas buriti (Mauritia flexuosa) vegetable oil (BVO) was non-cytotoxic to the cells. This study demonstrates that the characteristics of the BC structure can be modified, while preserving its intrinsic features, offering new possibilities for the development of BC-derived materials for specific applications in the biomedical field.
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Celulose , Óleos Voláteis , Óleos de Plantas , Celulose/química , Celulose/farmacologia , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Óleos de Plantas/química , Óleos de Plantas/farmacologia , Géis/química , Cicatrização/efeitos dos fármacos , Fabaceae/química , Humanos , Fibroblastos/efeitos dos fármacos , Pelargonium/química , Silanos/químicaRESUMO
OBJECTIVE: Oral candidiasis is a common fungal infection that affects the oral mucosa, and happens when Candida albicans interacts with bacteria in the oral microbiota, such as Streptococcus mutans, causing severe early childhood caries. C. albicans and S. mutans mixed biofilms are challenging to treat with conventional antimicrobial therapies, thus, new anti-infective drugs are required. This study aimed to test a drug delivery system based on chitosan microparticles loaded with geranium and lemongrass essential oils to inhibit C. albicans and S. mutans mixed biofilms. METHODOLOGY: Chitosan microparticles loaded with essential oils (CM-EOs) were obtained by spray-drying. Susceptibility of planktonic were performed according CLSI at 4 to 2,048 µg/mL. Mixed biofilms were incubated at 37ºC for 48 h and exposed to CM-EOs at 256 to 4,096 µg/mL. The antimicrobial effect was evaluated using the MTT assay, with biofilm architectural changes analyzed by scanning electron microscopy. RAW 264.7 cell was used to evaluate compound cytotoxicity. RESULTS: CM-EOs had better planktonic activity against C. albicans than S. mutans. All samples reduced the metabolic activity of mixed C. albicans and S. mutans biofilms, with encapsulated oils showing better activity than raw chitosan or oils. The microparticles reduced the biofilm on the slides. The essential oils showed cytotoxic effects against RAW 264.7 cells, but encapsulation into chitosan microparticles decreased their toxicity. CONCLUSION: This study demonstrates that chitosan loaded with essential oils may provide an alternative method for treating diseases caused by C. albicans and S. mutans mixed biofilm, such as dental caries.
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Quitosana , Cárie Dentária , Óleos Voláteis , Pré-Escolar , Humanos , Óleos Voláteis/farmacologia , Candida albicans , Streptococcus mutans , Quitosana/farmacologia , Cárie Dentária/prevenção & controle , BiofilmesRESUMO
Abstract Oral candidiasis is a common fungal infection that affects the oral mucosa, and happens when Candida albicans interacts with bacteria in the oral microbiota, such as Streptococcus mutans, causing severe early childhood caries. C. albicans and S. mutans mixed biofilms are challenging to treat with conventional antimicrobial therapies, thus, new anti-infective drugs are required. Objective This study aimed to test a drug delivery system based on chitosan microparticles loaded with geranium and lemongrass essential oils to inhibit C. albicans and S. mutans mixed biofilms. Methodology Chitosan microparticles loaded with essential oils (CM-EOs) were obtained by spray-drying. Susceptibility of planktonic were performed according CLSI at 4 to 2,048 µg/mL. Mixed biofilms were incubated at 37ºC for 48 h and exposed to CM-EOs at 256 to 4,096 µg/mL. The antimicrobial effect was evaluated using the MTT assay, with biofilm architectural changes analyzed by scanning electron microscopy. RAW 264.7 cell was used to evaluate compound cytotoxicity. Results CM-EOs had better planktonic activity against C. albicans than S. mutans. All samples reduced the metabolic activity of mixed C. albicans and S. mutans biofilms, with encapsulated oils showing better activity than raw chitosan or oils. The microparticles reduced the biofilm on the slides. The essential oils showed cytotoxic effects against RAW 264.7 cells, but encapsulation into chitosan microparticles decreased their toxicity. Conclusion This study demonstrates that chitosan loaded with essential oils may provide an alternative method for treating diseases caused by C. albicans and S. mutans mixed biofilm, such as dental caries.
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Electrospinning technology was used to produced polyvinylpyrrolidone (PVP)-copper salt composites with structural differences, and their virucidal activity against coronavirus was investigated. The solutions were prepared with 20, 13.3, 10, and 6.6% w/v PVP containing 3, 1.0, 0.6, and 0.2% w/v Cu (II), respectively. The rheological properties and electrical conductivity contributing to the formation of the morphologies of the composite materials were observed by scanning electron microscopy (SEM). SEM images revealed the formation of electrospun PVP-copper salt ultrafine composite fibers (0.80 ± 0.35 µm) and electrosprayed PVP-copper salt composite microparticles (1.50 ± 0.70 µm). Energy-dispersive X-ray spectroscopy (EDS) evidenced the incorporation of copper into the produced composite materials. IR spectra confirmed the chemical composition and showed an interaction of Cu (II) ions with oxygen in the PVP resonant ring. Virucidal composite fibers inactivated 99.999% of coronavirus within 5 min of contact time, with moderate cytotoxicity to L929 cells, whereas the virucidal composite microparticles presented with a virucidal efficiency of 99.999% within 1440 min of exposure, with low cytotoxicity to L929 cells (mouse fibroblast). This produced virucidal composite materials have the potential to be applied in respirators, personal protective equipment, self-cleaning surfaces, and to fabric coat personal protective equipment against SARS-CoV-2, viral outbreaks, or pandemics.
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The molecular weight of chitosan (CS) may affect its physical properties and its ability to induce an appropriate host response. The biocompatibilities of CS membranes of low (LMWCS) and high (HMWCS) molecular weight were investigated by inserting these materials into the subcutaneous tissue of rats for 1-28 days and evaluating leukocyte infiltration, granulation tissue, fibrosis, arginase-1 immunostaining, as well as nuclear factor-κB (NF-κΒ) and fibroblast growth factor (FGF)-2 expressions. Both CS membranes induced a peak of leukocyte infiltration on the first day of insertion and stimulated granulation and fibrous tissue generation when compared to control. LMWCS induced more collagen deposition a week earlier, when compared to the control and HMWCS membrane. The membranes also increased arginase-1 immunostaining, a M2 macrophage marker. M2 macrophage is recognized as anti-inflammatory and pro-regenerative. NF-κB is an essential biomarker of the inflammatory process and induces the expression of several pro-inflammatory cytokines. The LMWCS membrane reduced inflammation, as indicated by a reduced nucleus/cytoplasm NF-κB ratio in surrounding tissue from days 7 to 14 when compared to control. On the first day, the expression of FGF-2, a biomarker of inflammatory resolution, was increased in the tissue of the LWMCS group, when compared with HMWCS, which was consistent with the type I collagen deposition. Thus, LWMCS was associated with a prior reduction of the inflammatory response and improved wound healing.
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Materiais Biocompatíveis/química , Materiais Biocompatíveis/toxicidade , Quitosana/química , Quitosana/toxicidade , Inflamação/induzido quimicamente , Animais , Arginase/metabolismo , Colágeno/metabolismo , Citocinas , Fator 2 de Crescimento de Fibroblastos/metabolismo , Fibrose , Tecido de Granulação/patologia , Inflamação/patologia , Leucócitos/patologia , Masculino , Peso Molecular , NF-kappa B/metabolismo , Ratos , Ratos Wistar , CicatrizaçãoRESUMO
The aim of the study was to produce and characterize chitosan microparticles loaded with essential oils (CMEOs), evaluate the essential oil (EO) release profile and the CMEOs' anti-Candida activity. The chitosan microparticles (CMs) loaded with lemongrass essential oil (LEO) and geranium essential oil (GEO) were produced by the spray-drying method and characterized regarding CMEO morphological and physicochemical parameters and EO encapsulation efficiency (EE) and release profile. The planktonic activity was quantified by broth microdilution, and the activity against biofilm was quantified by biomass formation measurement. The LEO and GEO compositions were analyzed by gas chromatography combined with mass spectrometry (GC/MS), finding the main components citral (83.17%) and citronellol (24.53%). The CMs and CMEOs showed regular distribution and spherical shape (1 to 15 µm), without any morphological and physical modifications after EO incorporation. EE% ranged from 12 to 39%. In vitro release tests demonstrated the EO release rates, after 144 h, were 33% and 55% in PBS and HCl media, respectively. The minimum inhibitory concentration (MIC) values for CMEOs were lower than for CMs and pure EOs (P < 0.05). The higher CMEO biofilm inhibition percentage demonstrates the efficiency of microparticles against Candida biofilm. These results indicate that CMEOs are promising compounds that have antibiofilm activity against C. albicans.
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Biofilmes/efeitos dos fármacos , Candida albicans/efeitos dos fármacos , Candida albicans/fisiologia , Quitosana/química , Composição de Medicamentos , Óleos Voláteis/farmacologia , Antifúngicos/farmacologia , Cromatografia Gasosa-Espectrometria de Massas , Geranium/química , Testes de Sensibilidade Microbiana , Tamanho da Partícula , Espectroscopia de Infravermelho com Transformada de Fourier , Eletricidade Estática , TermogravimetriaRESUMO
Essential oils (EOs) are bioactive compounds with therapeutic potential for use as alternatives or as support to conventional treatments. However, EOs present limitations, such as sensibility to environmental factors, which can be overcome through microencapsulation. The objective of this study was to produce, by spray drying, chitosan microparticles (CMs) loaded with EO of Lemongrass (Cymbopogon flexuosus), Geranium (Pelargonium x ssp) and Copaiba (Copaifera officinalis). Physicochemical and biological characterization of these microparticles showed that CMs presented spherical morphology, had an average size range of 2-3 µm with positive zeta potential (ZP) values, and enhanced thermal stability, compared to free EO. The encapsulation efficiency (EE) ranged from 4.8-58.6%, depending on the oil's properties. In vitro EO release from CMs was determined at different pHs, with 94% release observed in acid media. All microparticles were non-hemolytic at concentrations of up to 0.1 mg·mL-1. EOs and CMs presented acetylcholinesterase (AChE) inhibition activity (IC 50 ranged from 11.92 to 28.18 µg·mL-1). Geranium and Copaiba EOs presented higher toxicity against Artemia salina, and greater inhibition of acetylcholinesterase, indicating potential bioactivity for Alzheimer's disease (AD). Our findings demonstrate that CM systems may show promise for the controlled release of these EOs.
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Artemia/efeitos dos fármacos , Cápsulas/química , Quitosana/química , Inibidores da Colinesterase/farmacologia , Cymbopogon/química , Fabaceae/química , Óleos Voláteis/análise , Pelargonium/química , Animais , Sangue/efeitos dos fármacos , Inibidores da Colinesterase/toxicidade , Cymbopogon/toxicidade , Fabaceae/toxicidade , Hemólise , Temperatura Alta , Humanos , Concentração de Íons de Hidrogênio , Concentração Inibidora 50 , Microscopia Eletrônica de Varredura , Óleos Voláteis/química , Tamanho da Partícula , Pelargonium/toxicidade , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Sporotrichosis, caused by Sporothrix schenckii complex species, is the most prevalent subcutaneous mycosis in many areas of Latin America. Chitosan has been used as an antifungal agent; however the effects of the molecular weight (MW) of chitosan (i.e. high (HMW), medium (MMW) and low (LMW) molecular weight chitosan) on S. brasiliensis has not been well described, particularly on biofilms. Effects on the planktonic form activity of S. brasiliensis were quantified by broth microdilution, while anti-biofilm activity was quantified by measuring metabolic activity via XTT (2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino)carbonyl]-2H-tetrazolium hydroxide and biomass formation (crystal violet). The molecular weight of chitosan modulated its effect on the planktonic form of S. brasiliensis, presenting lower MIC values for LMW chitosan. With regards both the adhesive and mature phases of biofilm, the LMW chitosan reduced biomass and metabolic activity most effectively. This study confirms the effects of the molecular weight and deacetylation degree of chitosan on its antifungal properties for potentially pathogenic fungi.
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Biofilmes/efeitos dos fármacos , Quitosana/farmacologia , Sporothrix/efeitos dos fármacos , Quitosana/química , Humanos , Peso Molecular , Plâncton/efeitos dos fármacos , Sporothrix/crescimento & desenvolvimento , Esporotricose/tratamento farmacológico , Esporotricose/patologiaRESUMO
Chitosan is a naturally occurring polysaccharide obtained from chitin, present in abundance in the exoskeletons of crustaceans and insects. It has aroused great interest as a biomaterial for tissue engineering on account of its biocompatibility and biodegradation and its affinity for biomolecules. A significant number of research groups have investigated the application of chitosan as scaffolds for tissue regeneration. However, there is a wide variability in terms of physicochemical characteristics of chitosan used in some studies and its combinations with other biomaterials, making it difficult to compare results and standardize its properties. The current systematic review of literature on the use of chitosan for tissue regeneration consisted of a study of 478 articles in the PubMed database, which resulted, after applying inclusion criteria, in the selection of 61 catalogued, critically analysed works. The results demonstrated the effectiveness of chitosan-based biomaterials in 93.4% of the studies reviewed, whether or not combined with cells and growth factors, in the regeneration of various types of tissues in animals. However, the absence of clinical studies in humans, the inadequate experimental designs, and the lack of information concerning chitosan's characteristics limit the reproducibility and relevance of studies and the clinical applicability of chitosan.
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Materiais Biocompatíveis/química , Quitosana/química , Regeneração , Engenharia Tecidual , Alicerces Teciduais , Animais , Humanos , Reprodutibilidade dos TestesRESUMO
In recent years, great effort has been devoted to the development of biomaterials that come into contact with blood. The surfaces of these materials need to be of suitable mechanical strength, and present anti-thrombogenic and anti-calcification properties. Chitosan is a natural polymer that has attracted attention due to its potential to act as a biomaterial. However, chitosan contains amino groups in its structure that may promote thrombogenesis and calcification. A strategy to reduce these properties constitutes the introduction of sulfonate groups (R-SO3-) in the chitosan chain. Another interesting biopolymer with similar characteristics to those of heparin is carrageenan, which has sulfate groups in its structure. As such, we evaluated "in vitro" calcification and thrombogenic processes on surfaces of pristine and sulfonated chitosan and on polyelectrolyte complexes (PEC) of chitosan and carrageenan. Results indicate that PEC demonstrate significant reductions in calcification and thrombogenic potential, probably due to the presence of sulfonate groups in both the carrageenan and treated chitosan.
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Anticoagulantes , Plaquetas/metabolismo , Carragenina , Quitosana , Anticoagulantes/química , Anticoagulantes/farmacologia , Carragenina/química , Carragenina/farmacologia , Quitosana/química , Quitosana/farmacologia , Avaliação Pré-Clínica de Medicamentos , Feminino , Humanos , MasculinoRESUMO
The catalytic activities of calcium oxide obtained from natural sources (crab shell and eggshell) were characterized and evaluated in the transesterification of vegetable oil. These catalysts are mainly composed of calcium carbonate, which is partially converted into CaO after calcination (900°C for 2h). The catalysts have some advantages, such as abundant occurrence, low cost, porous structure, and nontoxic. The materials were characterized by XRD, FTIR, TG/DTG, CO2-TPD, XPS, SEM, and BET methods. The thermal treatment produces small particles of CaCO3 and CaO that are responsible for the catalytic activity. The conversion from triglycerides to methyl ester was not observed in transesterification carried out using natural crab shell and eggshell. Under optimized reaction conditions, the conversions to YFAME using the calcined catalysts were: crab shell (83.10±0.27 wt.%) and eggshell (97.75±0.02 wt.%). These results, showed that these materials have promising viability in transesterification for biodiesel production.