RESUMEN
Methicillin-resistant Staphylococcus aureus (MRSA) is one of the leading causes of skin and soft tissue infections worldwide. This microorganism has a wide range of antibiotics resistance, a fact that has made the treatment of infections caused by MRSA difficult. In this sense, antimicrobial photodynamic therapy (aPDT) with natural products has emerged as a good alternative in combating infections caused by antibiotic-resistant microorganisms. The objective of the present study was to evaluate the effects of aPDT with Brazilian green propolis against intradermal MRSA infection in a murine model. Initially, 24 Balb/c mice were infected intradermally in the ears with 1.5 × 108 colony-forming units of MRSA 43300. After infection, they were separated into 4 groups (6 animals per group) and treated with the vehicle, only Brazilian green propolis, only blue LED light or with the aPDT protocol (Brazilian green propolis + blue LED light). It was observed in this study that aPDT with Brazilian green propolis reduced the bacterial load at the site of infection. Furthermore, it was able to inhibit weight loss resulting from the infection, as well as modulate the inflammatory response through greater recruitment of polymorphonuclear cells/neutrophils to the infected tissue. Finally, aPDT induced an increase in the cytokines IL-17A and IL-12p70 in the draining retromaxillary lymph node. Thus, aPDT with Brazilian green propolis proved to be effective against intradermal MRSA infection in mice, reducing bacterial load and modulating the immune response in the animals. However, more studies are needed to assess whether such effects are repeated in humans.
Asunto(s)
Antiinfecciosos , Staphylococcus aureus Resistente a Meticilina , Fotoquimioterapia , Própolis , Humanos , Ratones , Animales , Própolis/farmacología , Modelos Animales de Enfermedad , Brasil , Fotoquimioterapia/métodos , Antiinfecciosos/farmacología , Antibacterianos/farmacología , Antibacterianos/químicaRESUMEN
Staphylococcus aureus is the primary cause of skin and soft tissue infections. Its significant adaptability and the development of resistance are the main factors linked to its spread and the challenges in its treatment. Antimicrobial photodynamic therapy emerges as a promising alternative. This work aimed to characterize the antimicrobial photodynamic activity of Brazilian green propolis, along with the key bioactive compounds associated with this activity. Initially, a scanning spectrometry was conducted to assess the wavelengths with the potential to activate green propolis. Subsequently, reference strains of methicillin-resistant Staphylococcus aureus (MRSA ATCC 43300) and vancomycin-intermediate Staphylococcus aureus (VISA ATCC 700699) were exposed to varying concentrations of green propolis: 1 µg/mL, 5 µg/mL, 10 µg/mL, 50 µg /mL and 100 µg/mL and were stimulated by blue, green or red LED light. Finally, high-performance liquid chromatography coupled with a diode array detector and tandem mass spectrometry techniques, along with classic molecular networking analysis, was performed to identify potential bioactive molecules with photodynamic activity. Brazilian green propolis exhibits a pronounced absorption peak and heightened photo-responsiveness when exposed to blue light within the range of 400 nm and 450 nm. This characteristic reveals noteworthy significant photodynamic activity against MRSA and VISA at concentrations from 5 µg/mL. Furthermore, the propolis comprises compounds like curcumin and other flavonoids sourced from flavone, which possess the potential for photodynamic activity and other antimicrobial functions. Consequently, Brazilian green propolis holds promise as an excellent bactericidal agent, displaying a synergistic antibacterial property enhanced by light-induced photodynamic effects.
Asunto(s)
Antiinfecciosos , Staphylococcus aureus Resistente a Meticilina , Fotoquimioterapia , Própolis , Staphylococcus aureus , Fármacos Fotosensibilizantes/farmacología , Própolis/farmacología , Staphylococcus aureus Resistente a Vancomicina , Brasil , Antibacterianos/farmacología , Antiinfecciosos/farmacología , Fotoquimioterapia/métodos , Pruebas de Sensibilidad MicrobianaRESUMEN
Shrimp is among the most sensitizing food allergens and has been associated with many anaphylaxis reactions. However, there is still a shortage of studies that enable a systematic understanding of this disease and the investigation of new therapeutic approaches. This study aimed to develop a new experimental model of shrimp allergy that could enable the evaluation of new prophylactic treatments. BALB/c mice were subcutaneously sensitized with 100 µg of shrimp proteins of Litopenaeus vannamei adsorbed in 1 mg of aluminum hydroxide on day 0, and a booster (100 µg of shrimp proteins only) on day 14. The oral challenge protocol was based on the addition of 5 mg/ml of shrimp proteins to water from day 21 to day 35. Analysis of shrimp extract content detected at least 4 of the major allergens reported to L. vannamei. In response to the sensitization, allergic mice showed significantly enhanced IL-4 and IL-10 production in restimulated cervical draining lymph node cells. High detection of serum anti-shrimp IgE and IgG1 suggested the development of allergies to shrimp while Passive Cutaneous Anaphylaxis assay revealed an IgE-mediated response. Immunoblotting analysis revealed that Allergic mice developed antibodies to multiple antigens present in the shrimp extract. These observations were supported by the detection of anti-shrimp IgA production in intestinal lavage samples and morphometric intestinal mucosal changes. Therefore, this experimental protocol can be a tool to evaluate prophylactic and therapeutic approaches.
Asunto(s)
Anafilaxia , Hipersensibilidad a los Alimentos , Animales , Ratones , Inmunoglobulina E , Alérgenos , Extractos VegetalesRESUMEN
Methicillin-resistant Staphylococcus aureus (MRSA) is one of the main pathogens that cause infections in diabetic individuals. In this paper, we report the outcomes of our investigation on the intradermal application of antimicrobial photodynamic therapy (PDT) with curcumin in an infection induced by MRSA ATCC 43300 strain in the ear of mice with Type 1 Diabetes Mellitus (T1DM). A solution containing 100 µg of curcumin was photoactivated ex vivo with a LED light (450 nm) delivering a fluency of 13.5 J/cm3. This solution was administered in the ear intradermally, at the same inoculum site as the MRSA ATCC 43300 strain (PDT Group). This study also included the use of two control groups (both infected): One was treated with saline and the other was treated with non-photoactivated curcumin. The animals were euthanized 24 h after these treatments and samples of draining lymph node and treated ear were collected for examination. The PDT group showed lower bacterial load in the draining lymph node when compared to the saline and curcumin groups (p-value <0.05) 24 h after treatment. In addition to bacterial load, the PDT group presented a higher concentration of nitrates and nitrites in the draining lymph node when compared to the saline and curcumin groups (p-value <0.001). Examining the infectious site, despite apparently having similar inflammatory cell recruitment compared with the control groups, the PDT group showed a profile with less intense activity in the myeloperoxidase expression when compared to the saline group (p-value <0.001). Additionally, the detected concentration of cytokines such as IL-1ß, IL-12, and IL-10 was significantly lower in the PDT group when compared to the saline group (p-value <0.01; p-value <0.05; p-value <0.05, respectively), thus presenting a less intense inflammatory response during infection resolution. Our pilot study showed for the first time the therapeutic potential of PDT using curcumin when administered intradermally in the treatment of infections caused by S. aureus in mice with T1DM.
Asunto(s)
Curcumina/uso terapéutico , Diabetes Mellitus Tipo 1/complicaciones , Staphylococcus aureus Resistente a Meticilina/aislamiento & purificación , Fotoquimioterapia , Enfermedades Cutáneas Bacterianas/tratamiento farmacológico , Infecciones Estafilocócicas/tratamiento farmacológico , Animales , Biomarcadores/metabolismo , Mediadores de Inflamación/metabolismo , Ratones , Ratones Endogámicos C57BL , Proyectos Piloto , Enfermedades Cutáneas Bacterianas/complicaciones , Enfermedades Cutáneas Bacterianas/microbiología , Infecciones Estafilocócicas/complicaciones , Infecciones Estafilocócicas/microbiología , EstreptozocinaRESUMEN
Staphylococcus aureus is one of the major microorganisms that cause human diseases, leading from mild skin infections to serious diseases. With the use of semi-synthetic penicillins, methicillin-resistant strains called Methicillin-resistant Staphylococcus aureus (MRSA) have emerged, whose resistance pattern extends to other beta-lactam antibiotics. It has already been shown that photodynamic therapy is capable of inactivating MRSA as the laser excites the photosensitizer responsible for transferring its electrons to the molecular oxygen, generating extremely reactive molecules, such as singlet oxygen, being these reactive components the chemicals that promote the bacterial clearance. Thus, the research aiming at the development of new photosensitizers becomes important, especially to increase the amount of therapeutic resources available for the treatment of persistent infections related to this bacterium. In this context, Myrciaria cauliflora is a plant that has antimicrobial action and there are no reports of the use of its crude extract as a photosensitizer in antimicrobial photodynamic therapy. In that way, this work conveys an innovative way of the use of M. cauliflora extract as a photosensitizer, comprising its use as an antimicrobial agent when activated by light, against S. aureus. In vitro tests were performed where it was observed that after the photoactivation with blue LED light, the extract presented an augment in its antimicrobial activity, together with production of singlet oxygen. In the model of intradermal infection in Balb/c mice, a reduction in bacterial load was also detected, with raised expression of TNF-α, IL-17A, and MPO. Here, we demonstrate that the extract of M. cauliflora has photosensitizing action, promoting the production of singlet oxygen, besides IL-17A, TNF-α, and MPO.