Caracterização fenotípica e genotípica de bactérias recuperadas de implante ortopédico de conjunto placa-parafuso e parafuso de aço inoxidável austenítico após remoção cirúrgica
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Data
2011-08-29
Autores
Santos, Cássio Antonio Lanfredi dos [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
A maioria dos casos de fraturas ósseas é utilizada nas cirurgias, implantes ortopédicos de osteossíntese (placa-parafuso e parafusos) confeccionados em aço inoxidável austeníticos, devido à sua baixa relação custo-benefício. As infecções associadas aos implantes de osteossintese estão relacionadas com o crescimento de microrganismos em biofilmes, resultando em um processo infeccioso de difícil erradicação. O objetivo do estudo foi identificar os microrganismos recuperados do conjunto metálico placaparafuso e parafusos após remoção cirúrgica, avaliar a capacidade de aderência dos microrganismos, verificar a viabilidade celular, caracterizar genotipicamente os isolados Gram-positivos e detectar a resistência aos antimicrobianos. Os conjuntos placa-parafuso e parafusos de aço inoxidável austenítico ASTM F138/F139 e ISO NBR 5832-1/9 foram transportados em bolsa de polietileno para o Laboratório de Microbiologia Clínica. Os implantes foram lavados em solução tampão fosfato-salino, armazenados em bolsa contendo solução Ringer e submetidos ao banho ultrassônico em freqüência de 40±2 kHz por 5 minutos. O fluido sonicado foi transferido para tubos Falcon e centrifugado a 3.000g durante 20 minutos. O sedimento foi resuspendido em nova solução Ringer, homogeneizado por vortex e 10μL semeados em ágar Sangue de carneiro 5%, MacConkey e Sabouraud. Os meios foram incubados em diferentes condições de temperatura por 7 a 14 dias para recuperação de microrganismos. O perfil de resistência dos isolados foi obtido de acordo com CLSI 2011. Para análise da viabilidade celular foi utilizado o kit Live/Dead e microscópio de fluorescência...
The majority of cases of bone fractures is used in surgery, internal fixation of orthopedic implants (screw-plate and screws) manufactured in austenitic stainless steel due to its low cost-benefit. Infections associated with implant fixation are related to the growth of microorganisms in biofilms, resulting in an infection difficult to eradicate. The objective of study was to identify the microorganisms recovered from the set screwplate and screws metallic after surgical removal, evaluate the capacity of adherence of microorganisms, check the cell viability and characterize the isolates genotypically Gram-positive and detect antimicrobial resistance. The sets screw-plate and screws of austenitic stainless steel ASTM F138/F139 and ISO NBR 5832-1/9 were transported in polyethylene bag to the Laboratory of Clinical Microbiology. The implants were washed in phosphate buffered saline solution, stored in bags containing Ringer's solution and submitted to ultrasonic bath at a frequency of 40 kHz ± 2 for 5 minutes. The sonicate fluid was transferred to falcon tubes and centrifuged at 3.000g for 20 minutes The new sediment was re-suspended in Ringer's solution, homogenized by vortex and 10μL seeded agar 5% sheep blood, MacConkey and Sabouraud. The media were incubated at different temperatures for up to 7 days for growth of CFU mL-1. The resistance profile of the isolates was obtained according to CLSI 2011. For analysis of cell viability kit was used for Live/Dead and fluorescence microscope. After microbiological analysis the isolation was observed in some samples of polymicrobial implants. The data obtained showed that bacteria were the most isolated coagulase-negative... (Complete abstract click electronic access below)
The majority of cases of bone fractures is used in surgery, internal fixation of orthopedic implants (screw-plate and screws) manufactured in austenitic stainless steel due to its low cost-benefit. Infections associated with implant fixation are related to the growth of microorganisms in biofilms, resulting in an infection difficult to eradicate. The objective of study was to identify the microorganisms recovered from the set screwplate and screws metallic after surgical removal, evaluate the capacity of adherence of microorganisms, check the cell viability and characterize the isolates genotypically Gram-positive and detect antimicrobial resistance. The sets screw-plate and screws of austenitic stainless steel ASTM F138/F139 and ISO NBR 5832-1/9 were transported in polyethylene bag to the Laboratory of Clinical Microbiology. The implants were washed in phosphate buffered saline solution, stored in bags containing Ringer's solution and submitted to ultrasonic bath at a frequency of 40 kHz ± 2 for 5 minutes. The sonicate fluid was transferred to falcon tubes and centrifuged at 3.000g for 20 minutes The new sediment was re-suspended in Ringer's solution, homogenized by vortex and 10μL seeded agar 5% sheep blood, MacConkey and Sabouraud. The media were incubated at different temperatures for up to 7 days for growth of CFU mL-1. The resistance profile of the isolates was obtained according to CLSI 2011. For analysis of cell viability kit was used for Live/Dead and fluorescence microscope. After microbiological analysis the isolation was observed in some samples of polymicrobial implants. The data obtained showed that bacteria were the most isolated coagulase-negative... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Fraturas - Fixação interna, Biofilme, Infecção, Gene amplification icaA, Ultrasonic bath, Biofilm, Implant of osteosynthesis, Bacterial resistance
Como citar
SANTOS, Cássio Antonio Lanfredi dos. Caracterização fenotípica e genotípica de bactérias recuperadas de implante ortopédico de conjunto placa-parafuso e parafuso de aço inoxidável austenítico após remoção cirúrgica. 2011. 218 f. Dissertação (mestrado) - Universidade Estadual Paulista, Faculdade de Ciências Farmacêuticas, 2011.