4 research outputs found
Chitosan microparticles loaded with essential oils inhibit duo-biofilms of Candida albicans and Streptococcus mutans
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 ”g/mL. Mixed biofilms were incubated at 37ÂșC for 48 h and exposed to CM-EOs at 256 to 4,096 ”g/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
Chitosan microparticles loaded with essential oils inhibit duo-biofilms of Candida albicans and Streptococcus mutans
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 ”g/mL. Mixed biofilms were incubated at 37ÂșC for 48 h and exposed to CM-EOs at 256 to 4,096 ”g/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
Evaluation of citoprotector effect of amifostine on the doxorubicin-induced acute cardiotoxicity.
IntroduĂĂo: A Doxorubicina (DOX), antineoplĂsico antracĂclico, Ă largamente utilizada no tratamento dos mais diversos tipos de tumores sĂlidos e neoplasias hematolĂgicas. A Cardiotoxicidade provocada pelo uso de antibiĂticos antracĂclicos tem sido observada hĂ algumas dĂcadas como fator agravante e limitante do uso terapĂutico da DOX. Apesar do conhecimento de inĂmeros fatores que podem mediar a induĂĂo da cardiotoxicidade pela DOX, os mecanismos fisiopatolĂgicos continuam nĂo esclarecidos. Objetivo: Avaliar o efeito da amifostina e glutationa na cardiotoxicidade aguda induzida por doxorubicina e estudar a morfologia do tecido cardĂaco de camundongos tratados com doxorubicina por microscopia de forĂa atĂmica. Material e MĂtodos: Camundongos C57BL/6 fĂmeas (n=8) foram tratados com doxorubicina (25mg/Kg i.p.) ou salina (0,2mL i.p.) e sacrificados 96 horas apĂs tratamento. Outro grupo experimental foi tratado com amifostina (AMF 25, 50 e 100 mg/Kg s.c.), glutationa (GLT 125, 250 e 500mg/Kg s.c.) ou salina 30 mim antes da injeĂĂo de doxorubicina, no caso da glutationa a administraĂĂo foi diĂria atĂ o dia do sacrifĂcio. Os parĂmetros analisados foram: eletrocardiograma, Ăndices cardĂaco e esplĂnico, dosagem de grupos sulfidrilas nĂo protĂicos, dosagem de CK e CK-MB, parĂmetros histolĂgicos, dosagem por ELISA de TNF-, IL-1 expressĂo por imunohistoquĂmica de TNF-, IL-1, iNOS, apoptose e anĂlise por microscopia de forĂa atĂmica. Resultados: O tratamento com AMF nas doses de 50 e 100mg/Kg e GLT 250 e 500 mg/Kg foi capaz de aumentar a percentagem de sobrevivĂncia dos animais que foram submetidos a cardiotoxicidade aguda induzida por DOX (25 mg/Kg) quando comparados com o grupo injetado somente com DOX. A AMF e GLT tambĂm foram capazes de prevenir, em comparaĂĂo ao grupo DOX (p<0,05), as alteraĂĂes nos valores eletrocardiogrĂficos, (aumento do QRS e QTc e diminuiĂĂo da amplitude de R), as alteraĂĂes nos Ăndices cardĂacos e esplĂnicos, a elevaĂĂo dos nĂveis sĂricos das enzimas CK e CK-MB, a reduĂĂo dos nĂveis de grupos sulfidrilas nĂo protĂicos no tecido cardĂaco e as alteraĂĂes histolĂgicas (degeneraĂĂo hidrĂpica e vacuolizaĂĂo, focos de hialinizaĂĂo de fibras cardĂacas, picnose e necrose) induzidas pela DOX (25mg/Kg). A DOX induziu aumento da marcaĂĂo imunohistoquĂmica para cĂlulas apoptĂticas e expressĂo de iNOS e diminuiu a expressĂo de TNF-. A AMF foi capaz de prevenir estas alteraĂĂes, sendo esta prevenĂĂo apenas discreta para a expressĂo de TNF-. A microscopia de forĂa atĂmica revelou alteraĂĂes morfolĂgicas nĂo vistas pela microscopia Ăptica e mostrou ser uma ferramenta valiosa na avaliaĂĂo de efeitos de drogas. ConclusĂo: Nossos resultados sugerem o efeito citoprotetor da amifostina pelo aumento da atividade da glutationa peroxidase no tecido cardĂaco e que esta se mostra tĂo eficiente quanto a droga de referencia dexrazoxane. A utilizaĂĂo da microscopia atĂmica introduz uma ferramenta de anĂlise comparativa em escala nanomĂtrica, tornando possĂvel observar a destruiĂĂo membranar cardĂaco condizente com dano oxidativo.Introduction: Doxorubicin (DOX) is an antineoplasic anthracyclic agent used on the treatment of several solid tumors and hematological cancers. DOX-induced cardiotoxicity has been studied for decades as a limiting factor on the anticancer therapy with this drug. Despite the current knowledge concerning the mechanisms of DOX-induced cardotoxicity, its pathophysiology is still not clear. Purpose: To evaluate of the amifostine and glutathione citoprotective effect on the DOX-induced acute cardiotoxicity and to study the morphology of cardiac tissue through the use of atomic force microscopy as a tool. Materials and Methods: C57BL/6 female mice were treated with doxorubicin (25mg/Kg i.p.) or saline (0,2mL i.p.) and sacrificed 96 hours after treatment. In another experimental setting, mice were given amifostine (AMF 25, 50 e 100 mg/Kg s.c.), glutathione (GLT 125, 250 e 500mg/Kg s.c.) or vehicle, 30 mim before the administration of DOX, except glutathione that was injected daily. Analitical parameters included: electrocardiograms, cardiac and spleen indices, non protein suphidrils groups levels, CK and CK-MB cardiac enzymes levels, histological analysis, cardiac levels of (TNF-, IL-1, iNOS) determined by ELISA, immunohistochemistry for TNF-, IL-1, iNOS, apoptosis and atomic force microscopy tissue analysis. Results: AMF (100mg/Kg) and GLT (250 e 500 mg/Kg) treatments were able of improve the survival rate of animals in spite of the injection of DOX (25 mg/Kg) in comparison to DOX-treated only group (p<0.05). A AMF e GLT were also able to prevent electrocardiographic changes (rising of QRS e QTc and reduced R amplitude), changes in the cardiac and spleen indices, the augmentation of blood levels of CK e CK-MB, reduction of non proteic suphidrils groups levels, and histological changes induced by DOX (25mg/Kg). DOX induced the augmentation of the immunostaining for apoptotic cells and iNOS what was prevented by the administration of amifostine. The atomic force microscopy reveals morphological changes on the tissue organizational structure which is not possible to be observed through optical microscopy. Conclusion: Our results suggest that the amifostine citoprotective effect on DOX-induced acute cardiotoxicity is due the rising of glutathione peroxidase activity in the cardiac tissue. The citoprotective effect of amifostine is as efficient as the reference drug dexrazoxane. The use of atomic force microscopy as a new pharmacological tool for comparative analysis in nanometric scale allow us to observe DOX-induced membrane destruction what is suggestive of oxidative stress process.IntroduĂĂo: A Doxorubicina (DOX), antineoplĂsico antracĂclico, Ă largamente utilizada no tratamento dos mais diversos tipos de tumores sĂlidos e neoplasias hematolĂgicas. A Cardiotoxicidade provocada pelo uso de antibiĂticos antracĂclicos tem sido observada hĂ algumas dĂcadas como fator agravante e limitante do uso terapĂutico da DOX. Apesar do conhecimento de inĂmeros fatores que podem mediar a induĂĂo da cardiotoxicidade pela DOX, os mecanismos fisiopatolĂgicos continuam nĂo esclarecidos. Objetivo: Avaliar o efeito da amifostina e glutationa na cardiotoxicidade aguda induzida por doxorubicina e estudar a morfologia do tecido cardĂaco de camundongos tratados com doxorubicina por microscopia de forĂa atĂmica. Material e MĂtodos: Camundongos C57BL/6 fĂmeas (n=8) foram tratados com doxorubicina (25mg/Kg i.p.) ou salina (0,2mL i.p.) e sacrificados 96 horas apĂs tratamento. Outro grupo experimental foi tratado com amifostina (AMF 25, 50 e 100 mg/Kg s.c.), glutationa (GLT 125, 250 e 500mg/Kg s.c.) ou salina 30 mim antes da injeĂĂo de doxorubicina, no caso da glutationa a administraĂĂo foi diĂria atĂ o dia do sacrifĂcio. Os parĂmetros analisados foram: eletrocardiograma, Ăndices cardĂaco e esplĂnico, dosagem de grupos sulfidrilas nĂo protĂicos, dosagem de CK e CK-MB, parĂmetros histolĂgicos, dosagem por ELISA de TNF-, IL-1 expressĂo por imunohistoquĂmica de TNF-, IL-1, iNOS, apoptose e anĂlise por microscopia de forĂa atĂmica. Resultados: O tratamento com AMF nas doses de 50 e 100mg/Kg e GLT 250 e 500 mg/Kg foi capaz de aumentar a percentagem de sobrevivĂncia dos animais que foram submetidos a cardiotoxicidade aguda induzida por DOX (25 mg/Kg) quando comparados com o grupo injetado somente com DOX. A AMF e GLT tambĂm foram capazes de prevenir, em comparaĂĂo ao grupo DOX (p<0,05), as alteraĂĂes nos valores eletrocardiogrĂficos, (aumento do QRS e QTc e diminuiĂĂo da amplitude de R), as alteraĂĂes nos Ăndices cardĂacos e esplĂnicos, a elevaĂĂo dos nĂveis sĂricos das enzimas CK e CK-MB, a reduĂĂo dos nĂveis de grupos sulfidrilas nĂo protĂicos no tecido cardĂaco e as alteraĂĂes histolĂgicas (degeneraĂĂo hidrĂpica e vacuolizaĂĂo, focos de hialinizaĂĂo de fibras cardĂacas, picnose e necrose) induzidas pela DOX (25mg/Kg). A DOX induziu aumento da marcaĂĂo imunohistoquĂmica para cĂlulas apoptĂticas e expressĂo de iNOS e diminuiu a expressĂo de TNF-. A AMF foi capaz de prevenir estas alteraĂĂes, sendo esta prevenĂĂo apenas discreta para a expressĂo de TNF-. A microscopia de forĂa atĂmica revelou alteraĂĂes morfolĂgicas nĂo vistas pela microscopia Ăptica e mostrou ser uma ferramenta valiosa na avaliaĂĂo de efeitos de drogas. ConclusĂo: Nossos resultados sugerem o efeito citoprotetor da amifostina pelo aumento da atividade da glutationa peroxidase no tecido cardĂaco e que esta se mostra tĂo eficiente quanto a droga de referencia dexrazoxane. A utilizaĂĂo da microscopia atĂmica introduz uma ferramenta de anĂlise comparativa em escala nanomĂtrica, tornando possĂvel observar a destruiĂĂo membranar cardĂaco condizente com dano oxidativo.Introduction: Doxorubicin (DOX) is an antineoplasic anthracyclic agent used on the treatment of several solid tumors and hematological cancers. DOX-induced cardiotoxicity has been studied for decades as a limiting factor on the anticancer therapy with this drug. Despite the current knowledge concerning the mechanisms of DOX-induced cardotoxicity, its pathophysiology is still not clear. Purpose: To evaluate of the amifostine and glutathione citoprotective effect on the DOX-induced acute cardiotoxicity and to study the morphology of cardiac tissue through the use of atomic force microscopy as a tool. Materials and Methods: C57BL/6 female mice were treated with doxorubicin (25mg/Kg i.p.) or saline (0,2mL i.p.) and sacrificed 96 hours after treatment. In another experimental setting, mice were given amifostine (AMF 25, 50 e 100 mg/Kg s.c.), glutathione (GLT 125, 250 e 500mg/Kg s.c.) or vehicle, 30 mim before the administration of DOX, except glutathione that was injected daily. Analitical parameters included: electrocardiograms, cardiac and spleen indices, non protein suphidrils groups levels, CK and CK-MB cardiac enzymes levels, histological analysis, cardiac levels of (TNF-, IL-1, iNOS) determined by ELISA, immunohistochemistry for TNF-, IL-1, iNOS, apoptosis and atomic force microscopy tissue analysis. Results: AMF (100mg/Kg) and GLT (250 e 500 mg/Kg) treatments were able of improve the survival rate of animals in spite of the injection of DOX (25 mg/Kg) in comparison to DOX-treated only group (p<0.05). A AMF e GLT were also able to prevent electrocardiographic changes (rising of QRS e QTc and reduced R amplitude), changes in the cardiac and spleen indices, the augmentation of blood levels of CK e CK-MB, reduction of non proteic suphidrils groups levels, and histological changes induced by DOX (25mg/Kg). DOX induced the augmentation of the immunostaining for apoptotic cells and iNOS what was prevented by the administration of amifostine. The atomic force microscopy reveals morphological changes on the tissue organizational structure which is not possible to be observed through optical microscopy. Conclusion: Our results suggest that the amifostine citoprotective effect on DOX-induced acute cardiotoxicity is due the rising of glutathione peroxidase activity in the cardiac tissue. The citoprotective effect of amifostine is as efficient as the reference drug dexrazoxane. The use of atomic force microscopy as a new pharmacological tool for comparative analysis in nanometric scale allow us to observe DOX-induced membrane destruction what is suggestive of oxidative stress process