RESUMO
INTRODUCTION: The use of contact lenses has progressively increased around the world, thereby increasing the risk of complications. The most serious complication is microbial keratitis (corneal infection) that can progress to a corneal ulcer. METHODS: Fourteen multipurpose contact lens solutions were tested on mature biofilms comprising Staphylococcus aureus, Pseudomonas aeruginosa, Serratia marcescens and Candida albicans, using the minimum disinfection times recommended by the manufacturers. The biofilm was induced in the lens case, and 24 h later, the solutions were added. Activity against planktonic and sessile cells was evaluated and quantified as colony forming units per millilitre. The minimum concentration for biofilm eradication was defined as a 99.9% reduction in viable cells. RESULTS: Although most solutions exhibited activity against planktonic cells, only five of the 14 solutions produced a significant reduction in the S. marcescens biofilm. No solution achieved the minimal biofilm eradication of S. aureus, P. aeruginosa and C. albicans. CONCLUSION: Multipurpose contact lens solutions provide greater bactericidal and/or fungicidal activity on planktonic cells than biofilms. The minimal eradication biofilm concentration was only achieved for S. marcescens.
Assuntos
Soluções para Lentes de Contato , Staphylococcus aureus , Humanos , Soluções para Lentes de Contato/farmacologia , Pseudomonas aeruginosa , Candida albicans , Serratia marcescens , BiofilmesRESUMO
Infectious keratitis is a vision-threatening microbial infection. The increasing antimicrobial resistance and the fact that severe cases often evolve into corneal perforation necessitate the development of alternative therapeutics for effective medical management. Genipin, a natural crosslinker, was recently shown to exert antimicrobial effects in an ex vivo model of microbial keratitis, highlighting its potential to serve as a novel treatment for infectious keratitis. This study aimed to evaluate the antimicrobial and anti-inflammatory effects of genipin in an in vivo model of Staphylococcus aureus (S. aureus) and Pseudomonas aeruginosa (P. aeruginosa) keratitis. Clinical scores, confocal microscopy, plate count, and histology were carried out to evaluate the severity of keratitis. To assess the effect of genipin on inflammation, the gene expression of pro- and anti-inflammatory factors, including matrix metalloproteinases (MMPs), were evaluated. Genipin treatment alleviated the severity of bacterial keratitis by reducing bacterial load and repressing neutrophil infiltration. The expression of interleukin 1B (IL1B), interleukin 6 (IL6), interleukin 8 (IL8), interleukin 15 (IL15), tumor necrosis factor-α (TNF-α), and interferon γ (IFNγ), as well as MMP2 and MMP9, were significantly reduced in genipin-treated corneas. Genipin promoted corneal proteolysis and host resistance to S. aureus and P. aeruginosa infection by suppressing inflammatory cell infiltration, regulating inflammatory mediators, and downregulating the expression of MMP2 and MMP9.
Assuntos
Ceratite , Infecções por Pseudomonas , Humanos , Animais , Camundongos , Citocinas/metabolismo , Pseudomonas aeruginosa , Staphylococcus aureus/metabolismo , Projetos Piloto , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Ceratite/microbiologia , Córnea/metabolismo , Infecções por Pseudomonas/microbiologia , Camundongos Endogâmicos C57BLRESUMO
The purpose of this study was to investigate whether myofibroblast-related fibrosis (scarring) after microbial keratitis was modulated by the epithelial basement membrane (EBM) injury and regeneration. Rabbits were infected with Pseudomonas aeruginosa after epithelial scrape injury and the resultant severe keratitis was treated with topical tobramycin. Corneas were analyzed from one to four months after keratitis with slit lamp photos, immunohistochemistry for alpha-smooth muscle actin (α-SMA) and monocyte lineage marker CD11b, and transmission electron microscopy. At one month after keratitis, corneas had no detectible EBM lamina lucida or lamina densa, and the central stroma was packed with myofibroblasts that in some eyes extended to the posterior corneal surface with damage to Descemet's membrane and the endothelium. At one month, a nest of stromal cells in the midst of the SMA + myofibroblasts in the stroma that were CD11b+ may be fibrocyte precursors to myofibroblasts. At two to four months after keratitis, the EBM fully-regenerated and myofibroblasts disappeared from the anterior 60-90% of the stroma of all corneas, except for one four-month post-keratitis cornea where anterior myofibroblasts were still present in one localized pocket in the cornea. The organization of the stromal extracellular matrix also became less disorganized from two to four months after keratitis but remained abnormal compared to controls at the last time point. Myofibroblasts persisted in the posterior 10%-20% of posterior stroma even at four months after keratitis in the central cornea where Descemet's membrane and the endothelium were damaged. This study suggests that the EBM has a critical role in modulating myofibroblast development and fibrosis after keratitis-similar to the role of EBM in fibrosis after photorefractive keratectomy. Damage to EBM likely allows epithelium-derived transforming growth factor beta (TGFß) to penetrate the stroma and drive development and persistence of myofibroblasts. Eventual repair of EBM leads to myofibroblast apoptosis when the cells are deprived of requisite TGFß to maintain viability. The endothelium and Descemet's membrane may serve a similar function modulating TGFß penetration into the posterior stroma-with the source of TGFß likely being the aqueous humor.
Assuntos
Substância Própria/patologia , Úlcera da Córnea/patologia , Lâmina Limitante Posterior/fisiologia , Epitélio Corneano/fisiologia , Infecções Oculares Bacterianas/patologia , Infecções por Pseudomonas/patologia , Regeneração/fisiologia , Actinas/metabolismo , Animais , Biomarcadores/metabolismo , Antígeno CD11b/metabolismo , Lesões da Córnea/metabolismo , Lesões da Córnea/fisiopatologia , Substância Própria/metabolismo , Úlcera da Córnea/metabolismo , Modelos Animais de Doenças , Infecções Oculares Bacterianas/metabolismo , Feminino , Fibrose/patologia , Imuno-Histoquímica , Miofibroblastos/patologia , Infecções por Pseudomonas/metabolismo , CoelhosRESUMO
OBJETIVO: evaluar el comportamiento de las queratitis bacterianas con el tratamiento coadyuvante de suero autólogo tópico al 50 %. MÉTODOS: estudio comparativo, longitudinal y prospectivo. La muestra estuvo constituida por 60 pacientes, divididos de forma aleatoria en dos grupos: A) utilizó tratamiento antibiótico convencional y suero autólogo y B)utilizó solo tratamiento antibiótico convencional (cefazolina y amikacina). Los datos almacenados se procesaron utilizando el paquete estadístico SPSS 15. Las variables se expresaron según sus respectivas medidas de resumen y para la comparación de las terapias se aplicaron pruebas de hipótesis, con un nivel de confianza del 95 % y de error inferior al 0,05 %. RESULTADOS: predominaron en los aislamientos microbiológicos Staphylococcus y Pseudomonas; seguidas por los Streptococcus, gonococos y enterobacterias. En cuanto al tiempo de aparición de los signos que favorecen la cicatrización corneal y la respuesta terapéutica, encontramos que con la aplicación tópica del suero autólogo, en casi dos tercios de los pacientes, estos se manifestaron a partir de la segunda semana de tratamiento y se obtuvo una respuesta terapéutica favorable. CONCLUSIONES: el suero autólogo al 50 % resulta ser un complemento terapéutico efectivo en el manejo de las queratitis infecciosas de etiología bacteriana.
OBJECTIVE: to evaluate the behavior of bacterial keratitis with the adjuvant 50% topical autologous serum treatment. METHODS: comparative, longitudinal and prospective study of 60 patients, randomly distributed into two groups. Group A used the conventional antibiotics treatment plus autologous serum and Group B used only conventional antibiotics treatment (cephazolin and amykacin). The collected data were processed by the statistical package SPSS15. The variables were expressed according to their respective summary measures and hypothesis tests were applied to compare the two therapies; a confidence level of 95 % and error level less than 0,05 % were used. RESULTS: in the microbiological isolates, Staphylococcus and Pseudomonas prevailed, followed by Streptococcus, gonococci and Enterobacteriaceae. Regarding the time of onset of the signs that favor corneal healing and the therapeutic response, it was found that with the topical application of the autologous serum, almost two thirds of the patients showed healing signs since the second week of treatment, revealing a favorable therapeutic response. CONCLUSIONS: the 50 % autologous serum proves to be an effective therapeutic complement in the management of the infectious keratitis of bacterial etiology.
Assuntos
Humanos , Soluções Oftálmicas , Úlcera da Córnea/diagnóstico , Úlcera da Córnea/terapia , Soro/efeitos dos fármacos , Ceratite/terapia , Estudos de Casos e Controles , Estudos Prospectivos , Estudos LongitudinaisRESUMO
OBJETIVO: Avaliar o efeito do crosslinking (CXL) no tratamento de ceratite infecciosa, resistente ao tratamento clínico, e investigar a relação com o agente etiológico. MÉTODOS: Foram incluídos 11 pacientes com diagnóstico de ceratite infecciosa de etiologia bacteriana (sete olhos) e fúngica (quatro olhos) na Fundação Altino Ventura (FAV) no período de outubro de 2011 a maio de 2012. Os pacientes incluídos estavam em uso de colírios há pelo menos sete dias e não apresentavam melhora da infecção. Estes foram avaliados antes da realização do CXL e no período pós-operatório até cicatrização da úlcera. Para realização do CXL foram instiladas gotas de riboflavina a 0,1% e dextrano a 20%, a cada cinco minutos em um período de 30 minutos antes do procedimento, e durante a aplicação da luz ultravioleta A (UVA). A córnea foi exposta à UVA com comprimento de onda de 370ηm ± 5ηm e uma irradiância de 3mW/cm2. RESULTADOS: Os pacientes com infecção bacteriana obtiveram cura do processo infeccioso após o CXL e nenhum paciente com ceratite fúngica apresentou cicatrização. Observou-se associação significante (p = 0,003) entre o agente etiológico e a cicatrização. CONCLUSÃO: O CXL mostrou-se eficaz no tratamento da ceratite bacteriana resistente ao tratamento clínico, evitando a realização de transplante tectônico. Em relação à ceratite fúngica, este procedimento não influenciou na melhora do processo infeccioso.
PURPOSE: To evaluate the effect of corneal crosslinking (CXL) in the treatment of infectious keratitis resistant to medical treatment, and investigate the relation with the CXL outcome to the etiologic agent. METHODS: The study included 11 patients who were diagnosed with bacterial (seven eyes) or fungal keratitis (four eyes) at Altino Ventura Foundation from october 2011 to may 2012. All patients were using antibiotic eye drops for at least 7 days and have had no infection improvement. Patients were evaluated prior to CXL and the postoperative period until healing of the keratitis. For CXL, eyes were first instilled with a solution containing 0.1% riboflavin and 20% dextran for 30 min at a 5-minutes interval. Riboflavin-soaked eyes were then irradiated with UVA light (370ηm ± 5ηm) at 3mW/cm2 for 30 minutes. RESULTS: Eyes with bacterial infection exhibited improvement of infectious symptoms after CXL whereas eyes with fungal keratitis showed no improvement. Thus, there was a statistically significant correlation (p = 0.003) between the etiologic agent and the effectiveness of healing. CONCLUSION: CXL was effective in the treatment of bacterial keratitis resistant to clinical treatment, eliminating the need for surgery. However, CXL was not effective in managing fungal keratitis.
Assuntos
Humanos , Masculino , Feminino , Adolescente , Adulto Jovem , Pessoa de Meia-Idade , Ceratite/tratamento farmacológico , Infecções Oculares Bacterianas/tratamento farmacológico , Infecções Oculares Fúngicas/tratamento farmacológico , Reagentes de Ligações Cruzadas/uso terapêutico , Úlcera da Córnea/diagnóstico , Úlcera da Córnea/tratamento farmacológico , Antibacterianos/uso terapêutico , Antifúngicos/uso terapêutico , Acuidade VisualRESUMO
Avaliou-se a concentração de ofloxacina liberada por uma lente de contato de membrana de celulose biossintética, para tratamento de ceratite bacteriana experimental em cães, pela inoculação de Staphylococcus aureus intraestromal. Comparou-se o tratamento com a lente de contato biossintética impregnada com ofloxacina à terapia tópica convencional. Realizou-se avaliação microbiológica e dosagem de ofloxacina no humor aquoso por meio do método de cromatografia líquida de alto rendimento (HPLC). Houve diferença estatística na contagem de colônias bacterianas entre os olhos com ceratite e os demais grupos, no primeiro dia de coleta. O biomaterial, impregnado com ofloxacina, promoveu liberação gradual durante o período de avaliação, aos três e sete dias; no terceiro dia, o grupo tratado com a lente de contato obteve mediana de 3,72μg/mL, enquanto o grupo tratado com colírio resultou em 49,56μg/mL. Apesar do valor inferior, o grupo com lente de contato atingiu a concentração inibitória mínima, sendo eficaz no controle da infecção bacteriana.(AU)
The concentration of ofloxacin released by contact lens made of biosynthetic cellulose membrane was evaluated for the treatment of experimental bacterial keratitis in dogs by intrastromal inoculation of Staphylococcus aureus. The biosynthetic contact lens impregnated with ofloxacin was compared with the conventional topical therapy. The microbiological evaluation and the determination of ofloxacin in aqueous humor were evaluated by high performance liquid chromatography (HPLC). There was not statistical difference in the counting of bacterial colonies among the eyes with keratitis and other groups, on the first day of collection. The biomaterial, impregnated with ofloxacin, promoted gradual release during the evaluation period, at three and seven days; on the third day, the group treated with the contact lens obtained a median of 3.72μg/mL, while the group treated with eye drops resulted in 49.56μg/mL. Despite the lower value, the group with contact lens reached the minimum inhibitory concentration, which was effective in controlling the bacterial infection.(AU)
Assuntos
Animais , Cães , Fluoroquinolonas/administração & dosagem , Materiais Biocompatíveis , Lentes de Contato , Fluoroquinolonas/análise , Ceratite/tratamento farmacológico , Ceratite/veterinária , Cromatografia Líquida de Alta PressãoRESUMO
Avaliou-se a concentração de ofloxacina liberada por uma lente de contato de membrana de celulose biossintética, para tratamento de ceratite bacteriana experimental em cães, pela inoculação de Staphylococcus aureus intraestromal. Comparou-se o tratamento com a lente de contato biossintética impregnada com ofloxacina à terapia tópica convencional. Realizou-se avaliação microbiológica e dosagem de ofloxacina no humor aquoso por meio do método de cromatografia líquida de alto rendimento (HPLC). Houve diferença estatística na contagem de colônias bacterianas entre os olhos com ceratite e os demais grupos, no primeiro dia de coleta. O biomaterial, impregnado com ofloxacina, promoveu liberação gradual durante o período de avaliação, aos três e sete dias; no terceiro dia, o grupo tratado com a lente de contato obteve mediana de 3,72μg/mL, enquanto o grupo tratado com colírio resultou em 49,56μg/mL. Apesar do valor inferior, o grupo com lente de contato atingiu a concentração inibitória mínima, sendo eficaz no controle da infecção bacteriana.
The concentration of ofloxacin released by contact lens made of biosynthetic cellulose membrane was evaluated for the treatment of experimental bacterial keratitis in dogs by intrastromal inoculation of Staphylococcus aureus. The biosynthetic contact lens impregnated with ofloxacin was compared with the conventional topical therapy. The microbiological evaluation and the determination of ofloxacin in aqueous humor were evaluated by high performance liquid chromatography (HPLC). There was not statistical difference in the counting of bacterial colonies among the eyes with keratitis and other groups, on the first day of collection. The biomaterial, impregnated with ofloxacin, promoted gradual release during the evaluation period, at three and seven days; on the third day, the group treated with the contact lens obtained a median of 3.72μg/mL, while the group treated with eye drops resulted in 49.56μg/mL. Despite the lower value, the group with contact lens reached the minimum inhibitory concentration, which was effective in controlling the bacterial infection.