Lippia alba é uma espécie nativa, conhecida como erva cidreira , utilizada na medicina popular para o tratamento de distintas afecções. Estudos farmacológicos atribuem a sua atividade antimicrobiana aos terpenóides presentes no óleo essencial. Considerando a variação na composição de óleos essenciais de L. alba, o presente trabalho teve por objetivo determinar a composição química do óleo essencial de sete acessos de L. alba coletados no sul do Brasil, e avaliar a atividade antimicrobiana dos mesmos sobre um painel de 22 espécies bacterianas e 9 espécies de leveduras. Os óleos essenciais foram extraídos por hidrodestilação e analisados por Cromatografia gasosa e Cromatografia gasosa acoplada a um detector seletivo de massas. A atividade antimicrobiana (Dose de inibição 50% e Concentração inibitória mínima) foi determinada através do método de diluição seriada (0 a 10 mg/ml) em microplacas, com avaliação do crescimento por absorbância a 595nm. Para a determinação de Dose de inibição 50% e Concentração inibitória mínima foram consideradas as concentrações de óleo essencial que reduzidam o crescimento em 50% e 90%, respectivamente. A atividade bactericida e bacteriostática foi deteminada após 3h de tratamento dos microrganismos com a Concentração inibitória mínima pré-determinada para cada óleo. Com base nos seus componentes majoritários, os resultados permitiram caracterizar os sete acessos de L. alba como pertencentes aos quimiotipos: citral, 1,8-cineol/cânfora, linalol, linalol/mirceno/cânfora, carvona/cariofileno, linalol/1,8-cineol/germacreno D, e limoneno. Entre os quimiotipos, o óleo essencial citral exibiu o espectro antimicrobiano mais amplo, inibindo tanto bactérias como leveduras. Óleos com alta concentração de carvona/cariofileno e limoneno foram particularmente eficientes sobre leveduras, enquanto os quimiotipos linalol e linalol/1,8-cineol/germacreno D exibiram atividade inibitória sobre várias bactérias Gram positivas e negativas. Na Concentração inibitória mínima, o quimiotipo citral mostrou ação bactericida sobre leveduras e Salmonella sp. Entretanto, outros óleos apresentaram apenas efeito bacteriostático sobre a maioria dos microrganismos avaliados. Em geral, nossos resultados confirmam a atividade dos óleos essenciais de L. alba sobre microrganismos de importância clínica e industrial, mas apontam a necessidade de avaliação adequada da composição dos mesmos afim de escolher aquele mais efetivo para o controle de um microrganismo específico. / Submitted by Marcelo Teixeira (mvteixeira@ucs.br) on 2014-06-04T18:06:34Z No. of bitstreams: 1 Dissertacao Fernanda Formolo.pdf: 1268224 bytes, checksum: 2cde3df8d5929f9192359c2c9beb98df (MD5) / Made available in DSpace on 2014-06-04T18:06:34Z (GMT). No. of bitstreams: 1 Dissertacao Fernanda Formolo.pdf: 1268224 bytes, checksum: 2cde3df8d5929f9192359c2c9beb98df (MD5) / Lippia alba is a native species, known as "erva cidreira", currently used in folk medicine for the treatment of several diseases. Pharmacologic studies attributed their antimicrobial activity to the terpenoids present in the essential oils. Considering the variation on essential oil composition among L. alba accessions, the present work aimed to determine the essential oil composition of seven accession of L. alba collected in South Brazil, and to evaluate their antimicrobial activity against a panel of 22 bacterial and 9 yeast species. Essential oils were extracted by hydrodistillation and analyzed by GC and GC/MS. Antimicrobial activity (ID50 and MIC) were determined by a serial dilution method (0 to 10 mg/ml) on microplates with growth evaluation by absorbance at 595nm. For ID50 and MIC determination were considered the essential oil concentrations that reduced microbial growth on 50% and 90%, respectively. Bactericidal and bacteriostatic activities were determined after 3h treatment of the microorganisms with their individual MIC of each essential oil. Based on their main constituents, the results allowed characterizing the seven accessions of L. alba as chemotypes: citral, 1,8-cineol/camphor, linalool, linalool/mircene/camphor, carvone/caryophyllene, linalool/1,8-cineol/germacrene D, and limonene. Among these chemotypes, citral essential oils showed the largest antimicrobial spectrum, inhibiting both bacteria and yeasts. Oils with high concentration of carvone/caryophyllene, and limonene, were particularly effective against yeasts, and oils from chemotypes linalool and linalool/1,8-cineol/germacrene D exhibited inhibitory activity against several Gram positive and negative bacteria. At the MIC, citral showed bactericide effect on yeasts and Salmolella. However, other oils showed just bacteriostatic effects on most microorganisms. In general, our results confirm the activity of L. alba essential oils on both clinical and industrial microorganisms, but point out the necessity to properly evaluate essential oil composition to choose those more effective for the control of a specific microorganism.

INTRODUÇÃO: A combinação de sepse e insuficiência renal com necessidade de diálise é bastante comum nas Unidades de Terapia Intensiva e esta situação tem elevada mortalidade. Um desafio neste cenário é prescrever a dose correta dos antimicrobianos para o tratamento destas infecções. Em pacientes críticos e hemodinamicamente instáveis que necessitam de terapia renal substitutiva, um dos métodos mais utilizados é a diálise contínua, entretanto, recentemente, tem-se utilizado a diálise de baixa eficiência - conhecida como SLED, da abreviação do inglês \"sustained low-efficiency dialysis\". Esta modalidade de terapia renal substitutiva combina características da hemodiálise contínua com a intermitente, utilizando o equipamento da diálise intermitente, com menores fluxos sanguíneos e de dialisato, e com vantagem de menor custo. Apesar do fluxo mais baixo, por ser utilizado tempo mais prolongado, a SLED frequentemente resulta em maior clearance e especula-se que a remoção dos fármacos seria maior. Há escassez de estudos que avaliaram a farmacocinética e farmacodinâmica de antimicrobianos em pacientes submetidos à SLED.OBJETIVOS: Avaliar adequação farmacodinâmica de meropenem e vancomicina em pacientes submetidos a diálise estendida de baixa eficiência. Avaliar a depuração paramêtros farmacocinéticos durante a sessão de SLED. MÉTODOS: Foi realizado estudo prospectivo descritivo observacional com coleta de material biológico julho de 2012 a julho de 2014 HC-FMUSP. Foram incluídos pacientes submetidos à SLED em uso de vancomicina e/ou meropenem. Foram coletadas amostras de sangue seriadas (tempos: imediatamente antes do início da sessão de diálise, 0,5h, 1h, 2h, 4h após o início do tratamento e ao final da sessão). A quantificação dos antimicrobianos foi realizada através dos métodos analíticos de quantificação em Cromatografia Líquida de Alta Eficiência (CLAE). Os parâmetros farmacocinéticos foram calculados apenas durante a sessão de diálise utilizando-se o software WinNonlin. A área sob a curva foi determinada para a vancomicina. Para o meropenem, calculou-se o tempo acima da MIC. Resultados: Foram incluídos 24 pacientes tratados com vancomicina e 21 com meropenem eforam obtidas 170 amostras de plasma. As concentrações médias de vancomicina sérica e meropenem: antes da sessão de SLED foram 24,5 e 28,0 ?g / ml, respectivamente; e após SLED 14 e 6 ?g / ml, respectivamente. A depuração média foi de 41% para a vancomicina e 78% para o meropenem. Para vancomicina, 22 (96%), 19 (83%) e 16 (70%) pacientes teriam atingido o alvo (AUC0-24 > 400) considerando-se MIC 0,5; <= 1mg/l e <= 2 mg/l respectivamente. Para meropenem, 19 (95%), 18 (90%) e 11 (55%) pacientes teriam atingido a meta (70% de tempo acima da CIM) se infectados com isolados com MIC <= 1, <= 4 e <= 8 mg/l, respectivamente. Conclusões: Em pacientes críticos, meropenem evancomicina foram removidas durante o SLED. Entretando, a maioria dos pacientes alcançaria alvo PK-PD, excepto para CIMs mais altas. Sugerimos doses de manutenção de 1g a cada 12 ou 8 horaspara meropenem. Para a vancomicina, deve-se utilizar abordagem mais individualizada com monitorização sérica, uma vez que ensaios comerciais são disponíveis / Background: Antibiotic dosing is a challenge in critically ill patients undergoing renal replacement therapy. Our aim was to evaluate pharmacokinetics and pharmacodynamics of meropenem and vancomycin in patients undergoing SLED.Methods: ICU patients undergoing SLED, receiving meropenem and/or vancomycin, were prospectively evaluated. Blood samples were collected at the start of SLED and 0.5; 1; 2; 4 and 6 hours later. Antimicrobial levels were determined by HPLC. Noncompartimental pharmacokinetic analysis was performed. Area under the curve was determined for vancomycin. For meropenem, time above MIC was calculated. Results: 24 patients receiving vancomycin and 21 receiving meropenem were included; 170 plasma samples were obtained. Median serum vancomycin and meropenem concentrations: before SLED were 24.5 and 28.0 ?g/ml, respectively; and after SLED 14 and 6 ?g/ml, respectively. Mean removal was 41% for vancomycin and 78% for meropenem. For vancomycin, 22 (96%), 19(83%) and 16(70%) patients would have achieved the target (AUC0-24>400) considering MIC 0.5; <= 1mg/l and <= 2 mg/l, respectively. For meropenem, 19 (95%), 18 (90%) and 11(55%) patients would have achieved the target (70% of time above MIC) if infected with isolates with MIC <= 1, <= 4 and <= 8mg/l, respectively. Conclusions: In critically ill patients, meropenem and vancomycin were removed during SLED. Despite this, overall high PK/PD target attainment was obtained, except for higher MICs. We suggest maintenance doses of 1g tid or bid for meropenem. For vancomycin, more individualized approach using therapeutic drug monitoring should be used, as commercial assays are available

Lippia alba é uma espécie nativa, conhecida como erva cidreira , utilizada na medicina popular para o tratamento de distintas afecções. Estudos farmacológicos atribuem a sua atividade antimicrobiana aos terpenóides presentes no óleo essencial. Considerando a variação na composição de óleos essenciais de L. alba, o presente trabalho teve por objetivo determinar a composição química do óleo essencial de sete acessos de L. alba coletados no sul do Brasil, e avaliar a atividade antimicrobiana dos mesmos sobre um painel de 22 espécies bacterianas e 9 espécies de leveduras. Os óleos essenciais foram extraídos por hidrodestilação e analisados por Cromatografia gasosa e Cromatografia gasosa acoplada a um detector seletivo de massas. A atividade antimicrobiana (Dose de inibição 50% e Concentração inibitória mínima) foi determinada através do método de diluição seriada (0 a 10 mg/ml) em microplacas, com avaliação do crescimento por absorbância a 595nm. Para a determinação de Dose de inibição 50% e Concentração inibitória mínima foram consideradas as concentrações de óleo essencial que reduzidam o crescimento em 50% e 90%, respectivamente. A atividade bactericida e bacteriostática foi deteminada após 3h de tratamento dos microrganismos com a Concentração inibitória mínima pré-determinada para cada óleo. Com base nos seus componentes majoritários, os resultados permitiram caracterizar os sete acessos de L. alba como pertencentes aos quimiotipos: citral, 1,8-cineol/cânfora, linalol, linalol/mirceno/cânfora, carvona/cariofileno, linalol/1,8-cineol/germacreno D, e limoneno. Entre os quimiotipos, o óleo essencial citral exibiu o espectro antimicrobiano mais amplo, inibindo tanto bactérias como leveduras. Óleos com alta concentração de carvona/cariofileno e limoneno foram particularmente eficientes sobre leveduras, enquanto os quimiotipos linalol e linalol/1,8-cineol/germacreno D exibiram atividade inibitória sobre várias bactérias Gram positivas e negativas. Na Concentração inibitória mínima, o quimiotipo citral mostrou ação bactericida sobre leveduras e Salmonella sp. Entretanto, outros óleos apresentaram apenas efeito bacteriostático sobre a maioria dos microrganismos avaliados. Em geral, nossos resultados confirmam a atividade dos óleos essenciais de L. alba sobre microrganismos de importância clínica e industrial, mas apontam a necessidade de avaliação adequada da composição dos mesmos afim de escolher aquele mais efetivo para o controle de um microrganismo específico. / Lippia alba is a native species, known as "erva cidreira", currently used in folk medicine for the treatment of several diseases. Pharmacologic studies attributed their antimicrobial activity to the terpenoids present in the essential oils. Considering the variation on essential oil composition among L. alba accessions, the present work aimed to determine the essential oil composition of seven accession of L. alba collected in South Brazil, and to evaluate their antimicrobial activity against a panel of 22 bacterial and 9 yeast species. Essential oils were extracted by hydrodistillation and analyzed by GC and GC/MS. Antimicrobial activity (ID50 and MIC) were determined by a serial dilution method (0 to 10 mg/ml) on microplates with growth evaluation by absorbance at 595nm. For ID50 and MIC determination were considered the essential oil concentrations that reduced microbial growth on 50% and 90%, respectively. Bactericidal and bacteriostatic activities were determined after 3h treatment of the microorganisms with their individual MIC of each essential oil. Based on their main constituents, the results allowed characterizing the seven accessions of L. alba as chemotypes: citral, 1,8-cineol/camphor, linalool, linalool/mircene/camphor, carvone/caryophyllene, linalool/1,8-cineol/germacrene D, and limonene. Among these chemotypes, citral essential oils showed the largest antimicrobial spectrum, inhibiting both bacteria and yeasts. Oils with high concentration of carvone/caryophyllene, and limonene, were particularly effective against yeasts, and oils from chemotypes linalool and linalool/1,8-cineol/germacrene D exhibited inhibitory activity against several Gram positive and negative bacteria. At the MIC, citral showed bactericide effect on yeasts and Salmolella. However, other oils showed just bacteriostatic effects on most microorganisms. In general, our results confirm the activity of L. alba essential oils on both clinical and industrial microorganisms, but point out the necessity to properly evaluate essential oil composition to choose those more effective for the control of a specific microorganism.

Seafood permits the transmission of many bacterial pathogens. In order to reconcile consumer demands with important safety standards, traditional means of regulating microbial spoilage and safety hazards in foods are combined with novel technologies. These include biological antimicrobial systems, such as the use of lactic acid bacteria (LAB) and/or their bacteriocins, such as Carnobacterium maltaromaticum CS526 and its bacteriocin piscicocin CS526. The aims of this study were to investigate the presence of Listeria monocytogenes in temperate seafood, namely fresh and smoked salmon, fresh and smoked haddock, and fresh mussels and oysters. Additionally, there was an aim to recover, characterise and use bacteriocin-like-substance to control Listeria monocytogenes in cold smoked haddock. Vibrio spp., Enterobacteriaceae representatives, total aerobic heterotrophic counts and Listeria monocytogenes were isolated from commercially prepared smoked and fresh Atlantic salmon, smoked and fresh haddock, live mussels and oysters using selective media and tryptone soya agar (TSA). Vibrio spp. occurred in high densities (>106 CFU gˉ1) in mussels and Enterobacteriaceae representatives were recorded at >106 CFU gˉ1 in fresh salmon. Total aerobic heterotrophic counts in fresh salmon, live mussels and oysters reached 107, > 107, and > 106 CFU gˉ1, respectively. Listeria monocytogenes was recorded at 5.0 x 104 CFU gˉ1 in mussels. In total sixty one bacterial isolates were recovered from the seafood examined. The results revealed 19 genera of bacteria, i.e. Acinetobacter, Aerococcus, Aeromonas, Bacillus, Brochothrix, Carnobacterium, Citrobacter, Corynebacterium, Enterobacter, Escherichia coli, Moraxella, Micrococcus, Pseudomonas, Psychrobacter, Serratia, Shewanella, Staphylococcus, Vibrio and Listeria. The prominent characteristics of fish spoilage isolates were demonstrated by the ability of the isolates to reduce trimethylamine oxide (TMAO) to trimethylamine, and to produce H₂S. Sh. baltica OS185, Aeromonas spp. HB-6, Sh. baltica, Sh. putrefaciens, A. hydrophila HX201006-3, A. salmonicida subsp. achromogenes, A. hydrophila, C. freundii, Enterobacter cloacae were strong producers of TMA and H₂S. The spoilage microorganisms were tested for potential pathogenicity. The result revealed that 6/15 of the spoilage microorganisms produced proteolytic, lecithinase, blood (β and α haemolysin) and elastinase activity, respectively, whereas 7/15 of the spoilage microorganisms showed lipolytic activity. Cell free supernatants, ammonium sulphate precipitated supernatants and semi-purified bacteriocin-like substances of Carnobacterium maltaromaticum MMF-32 and KOPRI 25789 producing strains isolated from commercially prepared smoked salmon were investigated for their potential antimicrobial activity against potentially pathogenic and food spoilage microorganisms. Generally, a broad spectrum of activity was revealed against potentially pathogenic and food spoilage microorganisms in vitro. Cold-smoked haddock treated with bacteriocin producing C. maltaromaticum MMF-32, C. piscicola A9b bacˉ phenotype nonbacteriocin producing strain a mutant of C. piscicola A9b bac+, cell free supernatants, ammonium sulphate precipitated supernatants and semi-purified bacteriocin-like substances was challenged with L. monocytogenes ATCC 19114 up to 103 CFU gˉ1, respectively. Samples were stored at 4 °C for 10 days. L. monocytogenes and total bacterial counts were determined along with changes in total volatile base nitrogen (TVBN) and biogenic amines production as well as texture, colour and odour. Although the study on anti-listerial effects of C. maltaromaticum MMF-32 was not successful, this organism did have a positive effect on retention of firmness and sensory perception in cold smoked haddock.

<p>Nesavesna upotreba antibiotika izazvala je razvoj antimikrobne rezistencije zabrinjavajućih razmera. Porast rezistencije prema konvencionalnim antimikrobicima nametnuo je potrebu pronalaska novih supstanci, koje će biti delotvorne protiv otpornih sojeva. U poslednje četiri decenije, dominira trend da se lekovi i lekovita sredstava prave na bazi prirodnih proizvoda, a naročito biljnog materijala. Hemijski sastav plodova porodice ruža (Rosaceae) upućuje na to da bi određene vrste ove porodice biljaka mogle ispoljavati antimikrobno dejstvo. Plodovi voća porodice Rosaceae istraživani u okviru ove disertacije su: malina (Rubus idaeus), kupina (Rubus fruticosus), tre&scaron;nja (Prunus avium), vi&scaron;nja (Prunus cerasus) i aronija (Aronia prunifolia). Predmet istraživanja ove doktorske disertacije je antimikrobno tj. antibakterijsko, antigljivično, antialgalno i antivirusno dejstvo sokova i ekstrakata plodova odabranog voća porodice Rosaceae gajenog i prikupljenog na teritoriji Srbije na &scaron;irok spektar humanih i animalnih patogenih mikroorganizama.Sokovi su dobijeni ceđenjem svežih plodova voća. Ekstrakti su dobijeni iz ostataka voća nakon ceđenja (tropa) metodom dvostruke etanolne ekstrakcije. Nakon ceđenja i ekstrakcije sprovođeno je uparavanje do suva sokova i ekstrakata. HPLC metodom je ispitano prisutstvo 23 hemijske komponente u svakom od sokova i ekstrakata. Antimikrobna analiza je sprovedena na 21 mikroorganizmu, od čega su 15 bile bakterije, 2 gljivice, 2 alge i 2 virusi. Od navedenih mikroorganizama 14 je bilo autohtonih izolata, a 7 referentnih sojeva. Ispitivanje antibakterijskog, antifungalnog i antialgalnog dejstva plodova odabranog voća je sprovedeno difuzionom i mikrodilucionom metodom. Analiza antivirusne aktivnosti plodova odabranog voća je sprovedena posmatranjem inhibicije citopatogenog efekta (CPE) virusnih čestica na ćelijsku kulturu. Sve studije su sprovedene u triplikatu. Rezultati hemijske analize su obrađeni statistikom metodom srednje vrednosti, uz računanje standardne devijacije. Eksperimentalno dobijeni podaci u okviru ispitivanja antibakterijskog, antigljivičnog i antialgalnog dejstva su uvr&scaron;teni u statističku analizu. Metodom aproksimacije je utvrđena tačna koncentracija soka ili ekstrakta koja daje željeni efekat na ispitivani mikroorganizam. Rezultati analize antivirusnog dejstva su statistički obrađeni metodom srednje vrednosti. Kako bi se utvrdio stepen povezanosti rezultata dobijenih u okviru antimikrobnih analiza sa hemijskim supstancama prisutnim u plodovima odabranog voća sprovedena je regresiona analiza za vaki pojedinačni slučaj. Antibakterisjko, antiungalno i antialgalno dejstvo sokova i ekstrakata je upoređeno sa istim dejstvima konvencionalnih antimikrobnih lekova. Najveću ukupnu količinu ispitivanih supstanci je sadržao ekstrakt kupine (3965,56 mg/ml), a najmanju ekstrakt tre&scaron;nje (161,44 mg/ml). Od svih ispitivanih hemijskih supstanci, samo su: antocijani, katehin, galna kiselina i vitamin C, prisutni u svim ispitivanim sokovima i ekstraktima. Sve ispitivane biljke sadrže veću količinu supstanci u ekstraktu, osim kod tre&scaron;nje, kod koje se veća količina supstanci nalazi u soku. U najvećoj meri su u sokovima i ekstraktima plodova biljaka prisutni antocijani i to najvi&scaron;e kod kupine od ekstrakata, a kod maline od sokova. Antimikrobno (antibakterijsko, antifungalno, antialgalno i antivirusno) dejstvo je dokazano za svaki od sokova i/ili ekstrakata plodova odabranog voća. Po prosečnim vrednostima rezultata dilucione analize antimikrobnog i dejstva na MDBK ćelije, protiv najvećeg broja mikroorganizama najdelotvorniji je bio ekstrakt kupine, a neaktivan ili najslabije aktivan (aktivan u najvećoj koncentraciji) sok tre&scaron;nje. Prosečne vrednosti antimikrobnog i dejstva na MDBK ćelije su bile u opsegu od 0,71 mg/ml do 100 mg/ml. Sve ispitivane supstance su pokazale antibakterijsku aktivnost u ispitivanim koncentracijama na sve ispitivane bakterije, osim soka maline na Escherichia-u coli i soka aronije na Klebsiella-u pneumoniae. Ekstrakti su bolje delovali u većini slučajeva, uz nekoliko izuzetaka. Sve ispitivane supstance su pokazale antfiungalnu aktivnost na Candida-u albicans ATCC 24433, osim soka aronije. Na autohtoni soj Candida-e albicans su delovali samo ekstrakti kupine i tre&scaron;nje. Ekstrakti su bolje delovali u većini slučajeva, osim kupine na Candida-u albicans ATCC 24433, u kojem je dejstvo soka bilo jednako sa dejstvom ekstrakta. Iako su skoro svi ispitivani sokvi i ekstrakti ispoljili antialgalno dejstvo, na Prototheca-u wickerhamii nisu delovali vi&scaron;nja i sok maline, dok sok aronije nije delovao antialgalno ni na jednu od ispitivanih Prototheca. Ekstrakti su bolje delovali u većini slučajeva, osim maline kod koje je, u oba slučaja, bolje delovao sok. Sve ispitivane supstance su pokazale antivirusnu aktivnost na Bovine herpesvirus i Bovine viral diarrhea virus, kao i toksičnost na MDBK ćelije, osim sokova maline i tre&scaron;nje, koji su bili neaktivni protiv Bovine herpesvirus-a. Ekstrakti su bolje delovali u većini slučajeva, osim tre&scaron;nje na MDBK ćelije i Bovine viral diarrhea virus, na koje je jače delovao sok. U potpunosti jednako delovanje soka i ekstrakta je uočeno kod aronije za sva tri dejstva i maline za toksičnost na MDBK ćelije. Pri ispitivanju difuzionom metodom antibiotik je ispoljio jače dejstvo u odnosu na sokove i ekstrakte, osim ekstrakata kupine, maline i vi&scaron;nje na Trueperella-u pyogenes. Pri ispitivanju dilucionom metodom, dejstvo ekstrakata i sokova na Gram pozitivne bakterije je bilo uporedivo sa dejstvom Streptomicina, dok je dejstvo antibiotika na Gram negativne bakterije bilo jače u odnosu na sokove i ekstrakte, izuzetev jačeg dejstva ekstrakta maline protiv Escherichia-e coli. Nistatin je sve antifungalne i antialgalne efekte na gljivice i alge obuhvaćene istraživanjem ispoljio u manjim koncentracijama u odnosu na ispitivane sokove i ekstrakte.Najpovezanije sa antibakterijskim dejstvom sokova i ekstrakata na odgovarajuće mikroorganizme su bile: kumarinska, ferulna i galna kiselina, katehin. Najveći procenat povezanosti sa antimikrobnom aktivnosti ploda imali su antocijani na Enterococcus faecalis (71,94%). Najmanje povezani sa antimikrobnom aktivnosti na odgovarajuće mikroorganizme su bili: hlorogenska kiselina, kempferol, kumarinska i siringinska kiselina, vitamin C, vanilinska kiselina, epikatehin, katehin, rutin, krisin, elaginska i p-hidroksibenzoeva kiselina. Saznanja dobijena u okviru ove disertacije pro&scaron;iruju obim znanja o antimikrobnom dejtvu voća i predstavljaju osnovicu za svrsishodniju primenu voća kako u ishrani tako i u terapiji. Međutim, dalja istraživanja u smislu pro&scaron;irenja spektra ispitivanih mikroorganizama, antimikrobnih studija pojedinačnih komponenti iz plodova i mehanizama dejstva su potrebna.</p> / <p>Nonconscientious use of antibiotics has caused the development of antimicrobial resistance of worrying proportions. The increase in resistance to conventional antimicrobials has imposed a need to find new substances that will be effective against resistant strains. Over the last four decades, the trend is to medicines and medicinal products be made on the basis of natural products, and especially plant material. The chemical composition of the Rose family (Rosaceae) fruits suggests that certain species of this family of plants may exhibit an antimicrobial effect. The fruits of the Rosaceae family studied in the framework of this dissertation are: Raspberry (Rubus idaeus), Blackberry (Rubus fruticosus), Cherry (Prunus avium), Cherry (Prunus cerasus) and Aronia (Aronia prunifolia). The subject of the research of this doctoral dissertation is antimicrobial ie. antibacterial, antifungal, antialgal and antiviral effects of juices and fruit extracts, of the selected fruit of the Rosaceae family collected on the territory of Serbia, on a wide range of human and animal pathogenic microorganisms. Juices were obtained by the process of squeezing fresh fruits. The extracts were obtained from residues of fruits after extraction (pomace) by the double ethanol extraction method. After squeezing and extraction, evaporation to dryness of juices and extracts was carried out. The presence of 23 chemical components in each of the juices and extracts was examined by the HPLC method. Antimicrobial analysis was carried out on 21 microorganisms, of which 15 were bacteria, 2 fungi, 2 algae and 2 viruses. 14 of these microorganisms were autochthonous isolates and 7 reference strains. The antibacterial, antifungal and antialgal effect of the selected fruit was tested by the diffusion and microdilution method. The analysis of the fruits antiviral activity was carried out by observing the inhibition of cytopathogenic effect (CPE) of viral particles per cell culture. All studies were conducted in triplicate. The results of the chemical analysis were processed by means of a mean value method, with calculation of the standard deviation. The experimentally obtained data within the antibacterial, antifungal and antialgal examination were included in the statistical analysis. The method of approximation was used in order to determine the exact concentration of the juice or extract, which gives the desired effect on the examined microorganisms. The results of the antiviral analysis were statistically processed using the mean value method. In order to determine the degree of correlation of the results obtained in the framework of antimicrobial analysis with the chemical substances present in the fruits of the selected fruit, regression analysis was carried out for each individual case. Antibacterial, antifungal and antialgic effects of juices and extracts are compared with he same effects of conventional antimicrobial drugs, Streptomycin and Nystatin. The largest total amount of test substances was contained by blackberry extract (3965.56 mg/ ml), and the smallest by extract of sweet cherry (161.44 mg/ ml). Of all the investigated chemicals, only: anthocyanins, catechins, gallic acids and vitamin C were present in all examined juices and extracts. All investigated plants contain a higher amount of substances in the extract, except for sweet cherries, in which a greater amount of substances is present in the juice. In the juices and extracts of fruits anthocyanins are present to the greatest extent, mostly in the blackberry extract, and in raspberry juices. According to the average values of the results of the dilution analysis of the antimicrobial and the effect on the MDBK cell, the most effective was the blackberry extract and the most active inactive (active in the highest concentration) cherry juice against the most numerous microorganisms. The average antimicrobial values and effects on the MDBK cells were in the range of 0.71 mg/ ml to 100 mg/ ml. All investigated substances showed antibacterial activity in the tested concentrations on all examined bacteria, except raspberry juice on Escherichia coli and aronia juice on Klebsiella pneumoniae. Extracts acted better in most cases, with few exceptions. All investigated substances showed antifungal activity on Candida albicans ATCC 24433, except aronia juice. On the autochthonous Candida albicans, only blackberry and sweet cherry extracts were active. Extracts worked better in most cases, except for blackberry on Candida albicans ATCC 24433, in which the effect of the juice was equally efficient as the extract. Although almost all of the examined juices and extracts exhibited an antialgal effect, the sour cherry and raspberry juice did not work on the Prototheca wickerhamii, while the aronia juice did not express antialgal activity to any of tested Prototheca isolates. Extracts worked better in most cases, apart from raspberries, wich juice was stronger in both cases. All investigated substances showed antiviral activity on Bovine herpesvirus and Bovine viral diarrhea virus, as well as the toxicity to MDBK cells, except for raspberry and cherry juices, that were inactive against Bovine herpesvirus. The extracts worked better in most cases, except sweet cherries on MDBK cells and Bovine viral diarrhea virus, whose juice had stronger effect. The equally effective action of juice and extract was observed in aronia for all three effects and raspberries for toxicity to MDBK cells. In the diffusion test, antibiotic showed a stronger effect than juices and extracts, except for extracts of blackberry, raspberry and sour cherries on Trueperella pyogenes. In a dilution study, the effect of extracts and juices on Gram-positive bacteria was comparable to that of Streptomycin, while the effect of the antibiotic on Gram negative bacteria was stronger than juices and extracts, with exception of the stronger effect of raspberry extract against Escherichia coli. Nystatin has expressed its all antifungal and antialgal effects on fungi and algae included in the study in smaller concentrations compared to the examined juices and extracts. The most consistent with the antibacterial effect of juices and extracts on the appropriate microorganisms were: coumarin, ferulan and gallic acid, catechin. The highest percentage of association with antimicrobial activity of the fruits was given by anthocyanins to Enterococcus faecalis (71.94%). The least related to the antimicrobial activity on the appropriate microorganisms were: chlorogenic acid, champferol, coumarin and saringic acid, vitamin C, vanillic acid, epicatechin, catechin, rutin, chrysin, elaginic and p-hydroxybenzoic acid. The knowledge gained through this dissertation expands the scope of knowledge on antimicrobial activity of fruits and forms the basis for more effective use of fruits in both therapy and nutrition. However, further research in terms of extending the range of examined microorganisms, antimicrobial studies of individual fruits components and mechanisms of action are required.</p>

Indiana University-Purdue University Indianapolis (IUPUI) / The purpose of this study was to investigate the antibacterial efficacy of 0.12-percent and 2.0-percent chlorhexidine gluconate (CHX) on eliminating Enterococcus faecalis from dentinal tubules, and whether this antibacterial effect was enhanced by heat. To date there have been no published articles that describe the heating of 2.0-percent CHX and its antimicrobial efficacy and clinical relevance towards E. faecalis within dentinal tubules in root canal systems. Ninety-five human extracted, single rooted, maxillary, anterior teeth were used to prepare dentin disk specimens. After proper sterilization, a 2.5-mm ISO-sized diameter lumen was prepared, and then the canals were filled with brain-heart infusion (BHI) broth infected with E. faecalis. The BHI was removed and the specimens in equally divided groups were rinsed with sterile saline and filled with saline, or 0.12 percent CHX or 2.0 percent CHX at ambient temperature (24°C) or experimental temperature (46°C) and incubated at oral temperature (37°C) or the experimental temperature (46°C), respectively. The specimens were frozen to -70˚C and pulverized in liquid nitrogen. Serial dilutions were prepared of 1:100 and 1:1000 and spiral plated on BHI agar plates in duplicate. They were incubated, and the number of bacterial colonies was recorded 24 hours later for data analysis. A two-way analysis of variance (ANOVA), with factors for solution, solution temperature, and the solution-by-temperature interaction was used to determine antibacterial efficacy. Pair-wise comparisons between groups were examined for significance using the Fisher’s Protected Least Significant Differences Method. The E. faecalis CFU were log-transformed to satisfy the assumptions required for the ANOVA. The results of this investigation demonstrated no statistically significant difference with the addition of heat to either test irrigation solution regarding the elimination of E. faecalis from dentinal tubules within the root canal system. There was a statistically significant difference in the antibacterial efficacy of CHX against E. faecalis in comparison with the concentration tested. A higher concentration of 2.0-percent CHX demonstrated a significantly higher antibacterial efficacy against E. faecalis compared with 0.12-percent CHX, and likewise with the saline control. It can be concluded that the use of a higher concentration of 2.0-percent CHX is advantageous as a final irrigation solution after copious amounts of NaOCl and EDTA have been utilized for effective antimicrobial efficacy and substantivity.

No / Infection is the major reason of GTR/GBR membrane failure in clinical application. In this work, we developed GTR/GBR nanofiber membranes with localized drug delivery function to prevent infection. Metronidazole (MNA), an antibiotic, was successfully incorporated into electrospun polycaprolactone (PCL) nanofibers at different concentrations (0, 1, 5, 10, 20, 30, and 40 wt% polymer). To obtain the optimum anti-infection membrane, we systematically investigated the physical-chemical and mechanical properties of the nanofiber membranes with different drug contents. The interaction between PCL and MNA was identified by molecular dynamics simulation. MNA released in a controlled, sustained manner over 2 weeks and the antibacterial activity of the released MNA remained. The incorporation of MNA improved the hydrophilicity and in vitro biodegradation rate of PCL nanofibers. The nanofiber membranes allowed cells to adhere to and proliferate on them and showed excellent barrier function. The membrane loaded with 30% MNA had the best comprehensive properties. Analysis of subcutaneous implants demonstrated that MNA-loaded nanofibers evoked a less severe inflammatory response than pure PCL nanofibers. These results demonstrate the potential of MNA-loaded nanofiber membranes as GTR/GBR membrane with antibacterial and anti-inflammatory function for extensive biomedical applications.

Yes / Infection is the major reason for guided tissue regeneration/guided bone regeneration (GTR/GBR) membrane failure in clinical application. In this work, we developed GTR/GBR membranes with localized drug delivery function to prevent infection by electrospinning of poly(ε-caprolactone) (PCL) and gelatin blended with metronidazole (MNA). Acetic acid (HAc) was introduced to improve the miscibility of PCL and gelatin to fabricate homogeneous hybrid nanofiber membranes. The effects of the addition of HAc and the MNA content (0, 1, 5, 10, 20, 30, and 40 wt.% of polymer) on the properties of the membranes were investigated. The membranes showed good mechanical properties, appropriate biodegradation rate and barrier function. The controlled and sustained release of MNA from the membranes significantly prevented the colonization of anaerobic bacteria. Cells could adhere to and proliferate on the membranes without cytotoxicity until the MNA content reached 30%. Subcutaneous implantation in rabbits for 8 months demonstrated that MNA-loaded membranes evoked a less severe inflammatory response depending on the dose of MNA than bare membranes. The biodegradation time of the membranes was appropriate for tissue regeneration. These results indicated the potential for using MNA-loaded PCL/gelatin electrospun membranes as anti-infective GTR/GBR membranes to optimize clinical application of GTR/GBR strategies.

<p>Najče&scaron;ća komplikacija nakon ugradnje silikonskih implantata za dojku je kontraktura fibrozne kapsule (KK), koja se normalno stvara oko implantata u sklopu reakcije oko stranog tela. Najozbiljnija komplikacija nakon ugradnje silikonskih implantata za dojku je anaplastični krupnoćelijski limfom koji se javlja isključivo kod pacijentkinja koje imaju ugraĎene implantate (eng. Breast-implant associated anaplastic large cell lymphoma &ndash; BIA ALCL). Uzrok nastanka ovih komplikacija ostaje nepoznat. Ustanovljeno je da se KK manje javlja kod implantata koji imaju makroteksturisanu povr&scaron;inu i kod onih koji su presvučeni poliuretanskom penom. S druge strane, BIA-ALCL se če&scaron;će dijagnostikuje kod pacijentkinja kojima su ugraĎeni upravo makroteksturisani implantati. Subklinička infekcija koja predstavlja odgovor organizma na postojanje biofilma na ugraĎenim implantatima, predstavlja jedan od najznačajnijih etiolo&scaron;kih faktora za nastanak KK i BIA-ALCL. Biofilm je konglomerat mirkoorganizama uronjenih u matriks koji ih &scaron;titi od dejstva antibiotika i antiseptika. Kako je nemoguće delovati medikamentozno na eradikaciju biofilma, brojni autori daju razne preporuke u cilju izbegavanja kontaminacije implantata tokom operativnog zahvata, a time i formiranja biofilma. Pored brojnih mera, savetuje se i ispiranje džepa u koji će se plasirati proteza kao i same proteze, nekim od antiseptičkih ili antibiotskih rastvora. Do sada ne postoje prihvaćene jasne preporuke o načinu ispiranja različitih implantata, objavljena su samo lična iskustva raznih autora. Ciljevi ovog istraživanja su bili da se ustanovi mogućnost formiranja biofilma četiri različite bakterije (Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa i Ralstonia pickettii) na tri različito teksturisana silikonska implantata za dojku (sa porama veličine 70-150 &mu;m, 50&ndash;900 &mu;m i 13 &mu;m) u in vitro uslovima; da se ispita da li ispiranje antisepticima (oktenidindihidrohloridom i povidon jodom), ili antibiotikom (cefuroksimom) ili istovremeno me&scaron;avinom povidon joda i antibiotika pre bakterijske kontaminacije sa četiri različite bakterije ima uticaja na formiranje biofilma na tri različito teksturisana implantata za dojke u in vitro uslovima; i da se ispita efekat antiseptika u odnosu na efekat antibiotika na formiranje bakterijskog biofilma na tri različito teksturisana silikonska implantata za dojku. Istraživanje je koncipirano kao prospektivna studija u vidu eksperimenta koji je izveden u Laboratoriji za mikrobiologiju, Instituta za javno zdravlje Vojvodine u Novom Sadu. Za izvoĎenje eksperimenta kori&scaron;ćeni su uzorci tri vrste silikonskih implantata za dojku sa različito teksturisanom povr&scaron;inom, odnosno porama različite veličine: 70-150 &mu;m, 50&ndash;900 &mu;m, i 13 &mu;m. Od svakog od navedenih implantata su pravljeni uzorci, sečenjem kapsula implantata na komadiće veličine 1x1 cm. Ukupno je bilo 1440 uzoraka. Na osnovu teksture uzorci su podeljeni u tri grupe: Grupa 1 (pore veličine 70-150 &mu;m), Grupa 2 (pore veličine 50&ndash;900 &mu;m) i Grupa 3 (pore veličine 13 &mu;m). Svaka od ovih grupa je dalje podeljena u jednu kontrolnu grupu i po četiri ispitivane grupe. Nakon sterilizacije uzoraka svaka kontrolna grupa je kontaminirana sa po 100&mu;l bakterijskog bujona Staphylococcus epidermidis (n=30), Staphylococcus aureus (n=30), Pseudomonas aeruginosa (n=30) i Ralstonia pickettii (n=30). Ispitivane grupe se bile podeljene prema načinima ispiranja na one u kojima su uzorci prvo ispirani: oktenidin &ndash; dihidrohloridom ili povidon jodom ili cefuroksimom ili kombinacijom povidon joda i dva antibiotika, pa potom kontaminirani sa po 100&mu;l bakterijskog bujona Staphylococcus epidermidis (n=30), Staphylococcus aureus (n=30), Pseudomonas aeruginosa (n=30) i Ralstonia pickettii (n=30). Po zavr&scaron;enoj kontaminaciji, uzorci su se inkubirali na temperaturi od 37&deg;C u trajanju od 96h, čime su stvoreni uslovi za formiranje biofilma. Nakon inkubacije, svaki pojedinačni uzorak je uronjen u sterilan tripton soja bujon, izlagan soničnoj energiji u trajanju od 1minuta i zatim vorteksiran 1 minut, čime je omogućeno odvajanje nastalog biofilma od implantata. Za ispitivanje sposobnosti formiranja biofilma kori&scaron;ćena je modifikovana tehnika sa mikrotitar pločom po Stepanoviću. Rezultati su pokazali da sve četiri ispitivane bakterije S. epidermidis, S. aureus, P. aeruginosa i Ralstonia pickettii statistički značajno vi&scaron;e stvaraju biofilm na implantatima sa porama veličine 50&ndash;900 &mu;m u odnosu na pore 70-150 &mu;m i u odnosu na pore veličine 13 &mu;m. Biofilm se statistički značajno vi&scaron;e stvara na porama veličine 70-150 &mu;m u odnosu na pore 13 &mu;m. Jedini izuzetak je Pseudomonas aeruginosa kod kojeg ne postoji statistični značajna razlika u produkciji biofilma na teksturisanim implantatima sa porama veličine 70-150 &mu;m u odnosu na one sa porama 13 &mu;m. TakoĎe, sve četiri ispitivane bakterije statistički značajano manje stvaraju biofilm nakon ispiranja povidon jodom, oktenidin-dihidrohloridom ili rastvorom antibiotika u sve tri grupe implantata, u odnosu na povr&scaron;ine koje nisu ispirane. Izuzetak je S. epidermidis u Grupi 3 kod kojeg nije utvrĎeno statistički značajno manje formiranje biofilma nakon ispiranja oktenidin dihidrohloridom u odnosu na neispiranje. Cefuroksim je bio efikasniji u sprečavanju formiranja biofilma sve četiri ispitivane bakterije u odnosu na neispiranje u Grupi 1, kao i za S. epidermidis i Ralstoniu Pickettii u Grupi 2. Cefuroksim se nije pokazao statistički značajno efikasnim u sprečavanju formiranja biofilma S. aureus i P. aeruginosa u Grupi 2, kao ni kod jedne bakterije u Grupi 3. Dalje je dokazano da su antiseptici (oktenidin-dihirohlorid i povidon jod) kao i me&scaron;avina povidon joda i dva antibiotika (cefuroksim i gentamicin), statistički značajno efikasnji od ispiranja samo antibiotikomcefuroksimom u smanjenju formiranja biofilma sve četiri ispitivane bakterije kod sva tri ispitivana, različito teksturisana silikonska implantata. Rezultati su pokazali da je ispiranje povidon jodom statistički značajno efikasnije u prevenciji stvaranja biofilma kod skoro svih ispitivanih bakterija od ispiranja oktenidin- dihidrohloridom u sve tri grupe implantata. Statistički značajna razlika nije utvrĎena u prevenciji stvaranja biofilma Staphylococcus aureusa kod sve tri grupe implantata prilikom ispiranja povidon jodom u odnosu na oktenidin- dihidrohlorid, kao i kod Ralsotnia pickettii u Grupi 2. Na osnovu rezultata ove studije, preporuka je da se koriste mikroteksturisani implantati kao i da se oni, pre ugradnje isperu povidon jodom ili me&scaron;avinom povidon jod i dva antibiotika (cefuroksim i gentamicin), u cilju prevencije stvaranja biofilma, a time i postoperativnih komplikacija koje mogu nastati nakon ugradnje implantata.</p> / <p>The most common complication after breast implant surgery is contracture of capsule, which is normally formed around implants as part of foreign body reaction. The most sincere complication after this kind of surgery is breast implant associated anaplastic large cell lymphoma (BIA-ALCL). The cause of these complications is still unknown. It is evident that capsular contracture (CC) is seen less frequently in patients with macro-textured implants and in those with implants covered with polyurethane foam. On the other hand, BIA-ALCL is diagnosed more frequently in patients with those, macro-textured implants. Subclinical infection, defined as an response of organism on presence of biofilm on the implant, is considered to be one of the most important etiologic factors for CC and BIA-ALCL. Biofilm is a conglomerate of microorganisms immersed into matrix, which protects them from influence of antibiotics and antiseptics. As it is impossible to eradicate biofilms with medicaments, many authors suggest different steps in order to avoid contamination of the implant during the operation and therefore, prevent the formation of biofilm. Among many tips, it is recommended to irrigate the pocket for breast implant and the implant itself, with some antiseptic or antibiotic solution. Up till now, there is no agreed consensus on the type of irrigation for different implants. Only personal experiences of a few authors have been published. Aims of this research were: to establish the possibility of biofilm formation of four different bacteria (Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa and Ralstonia pickettii) on three differently textured breast implants (with pore diameter of 70-150 &mu;m, 50&ndash;900 &mu;m and 13 &mu;m) in vitro; to examine whether the irrigation of implant with antiseptics (povidone iodine and octenidine dihydrochloride), antibiotics (cefuroxime) or mixture of povidone iodine and two antibiotics, before the contamination with bacteria, has an influence on the incidence on biofilm formation on three differently textured implants; and to examine the effect of antiseptics in contrast to the effect of antibiotics on biofilm formation on three differently textured breast implants. The study was conducted as a prospective research that took place at the Laboratory for microbiology, at the Institute of public health of Vojvodina in Novi Sad. For the experiment, three types of silicone breast implants were used with different pore sizes: 70-150 &mu;m, 50&ndash;900 &mu;m and 13 &mu;m. Samples were made by cutting each of these types of implants into pieces sized 1x1cm. There were 1440 samples in total. According to texture, samples were divided it three groups: Group 1 (pore size 70-150 &mu;m), Group 2 (pore size 50&ndash;900 &mu;m) and Group 3 (pore size 13 &mu;m). Furthermore, each of these groups was divided in one control and four test groups. After sterilisation of samples, every control group was contaminated with 100&mu;l of bacterial broth of Staphylococcus epidermidis (n=30), Staphylococcus aureus (n=30), Pseudomonas aeruginosa (n=30) and Ralstonia pickettii (n=30). Tested groups were divided according to type of irrigation into those where samples were firstly irrigated with either: octenidine dihydrochloride of povidone iodine or cefuroxime of mixture of povidone iodine with two antibiotics, and after the irrigation, contaminated with 100&mu;l bacterial broth of Staphylococcus epidermidis (n=30), Staphylococcus aureus (n=30), Pseudomonas aeruginosa (n=30) and Ralstonia pickettii (n=30). After contamination, samples were incubated on 37&deg;C for 96h, which created excellent conditions for biofilm formation. After incubation, each sample was dipped into sterile tripton soy broth, and then exposed to sonic energy for 1 minute and vortexed for 1 minute, which made biofilm separate from the implant. For testing the capability of biofilm formation, modified technique with microtitar plates described by Stepanović was used. Results show that all four examined bacteria S. epidermidis, S. aureus, P. aeruginosa and Ralstonia pickettii form more biofilm on implants with pore sizes 50&ndash;900 &mu;m compared to implants with pore size 70-150 &mu;m and those with 13 &mu;m. Statistical significance was found in biofilm formation on implants with pores 70-150 &mu;m compared to implants with pores 13 &mu;m. Furthermore, all four examined bacteria form statistically less biofilm after the irrigation with any of used solutions: povidone iodine, octenidine dihydrochloride, antibiotic solution of mixture of povidone iodine and two antibiotics, in all three groups of implants compared to surfaces that were not irrigated. The exception is S. epidermidis in Group 3, where no statistical significance was found on biofilm formation after the irrigation with octenidine dihydrochloride compared to non-irrigation. Cefuroxime was more efficient in biofilm prevention for all four tested bacteria compared to non-irrigation in Group 1 and for S. epidermidis and Ralstonia pickettii in Group 2. There was no statistical significance found in prevention of S. aureus i P. aeruginosa biofilms when irrigating with cefuroxime in Group 2, as well as for all tested bacteria in Group 3. Furthermore, it was verified that antiseptics (octenidin dihydrochloride and povidone iodine) and mixture of povidone iodine and two antibiotics (cefuroxime and gentamycin), were statistically more efficient in biofilm prevention of all four examined bacteria in all groups of implants, compared to irrigation with antibiotic-cefuroxime alone. Results show that irrigation with povidone iodine is statistically more efficient in biofilm prevention of almost all examined bacteria compared to irrigation with octenidine dihydrochloride in all groups of implants. There was not found any statistical significance in prevention of Staphylococcus aureus biofilm when irrigating with povidone iodine compared to octenidine dihydrochloride in all groups of implants, and also in biofilm prevention of Ralsotnia pickettii in Group 2. According to results of this research, it is recommended to use micro-textured implants and to irrigate them with povidone iodine or mixture of povidone iodine and two antibiotics (cefuroxime and gentamycin) prior the implementation, in order to prevent biofilm formation which is most probable cause of postoperative complications after implant surgery.</p>

El texto completo de este trabajo no está disponible en el Repositorio Académico UPC por restricciones de la casa editorial donde ha sido publicado. / PURPOSE: To assess the long-term effect of a 2% aqueous chlorhexidine (CHX) solution on bond strength to artificially eroded dentin compared to sound dentin. MATERIALS AND METHODS: Flat mid-coronal dentin surfaces of extracted third molars (n = 28) were subjected only to grinding with a 600-grit SiC paper for 1 min (sound dentin S, n = 14) or additionally to erosive pH cycling with a cola-based soft-drink (eroded dentin E, n = 14). After acid etching, rinsing, and air drying, S and E were rehydrated with 1.5 μl of 2% CHX (S2%, n = 7; E2%, n = 7) or of distilled water (control SC, n = 7; EC, n = 7). Composite buildups were incrementally constructed with Filtek Z350 following Adper Single Bond 2 application. Specimens were sectioned into beams, which were subjected to microtensile testing immediately or after 6 or 12 months of aging. Fractured surfaces were observed under a digital microscope (50X magnification). Microtensile bond strength (μTBS) (MPa) was analyzed by three-way ANOVA and Tukey's tests (α = 0.05) and failure mode by the Kruskal-Wallis test (α = 0.05). RESULTS: Compared to sound dentin, eroded dentin was consistently related to lower μTBS. Immediately and after 12-month aging, the effect of CHX was insignificant, but it was significant after 6-month aging, when it conserved the bond strength to both eroded and sound dentin. The percentage of adhesive and mixed failures were equivalent, and significantly more frequent than cohesive failures, whether in dentin or in composite. CONCLUSION: The 2% CHX effect on bond strength conservation to both eroded and sound dentin was not found to be persistent. / Revisión por pares