Toxic products, such as endotoxin from the gram-negative and exotoxin from the gram-positive bacteria, are the most important initiators of the inflammatory host response in sepsis. In addition to antibacterial treatment, numerous attempts have been made to interfere with the exaggerated proinflammatory cascade initiated by the toxins. As most antitoxic and anti-inflammatory agents have shown no clear efficacy, an attractive alternative has been to prevent or minimise their release. Therefore, it was of interest to further study the antibiotic-induced release of toxins after exposure to antibiotics used for the treatment of the most severe infections, especially if protein synthesis inhibitors could reduce the release induced by PBP 3-specific β-lactam antibiotics. There were significant reductions in endotoxin release from gram-negative bacteria when the combination of the PBP 3-specific β-lactam antibiotic, cefuroxime, and the protein synthesis inhibitor, tobramycin, was compared with cefuroxime alone. Increasing doses of tobramycin reduced endotoxin release and increased the killing rate. In a kinetic in vitro model the endotoxin release from E.coli was higher after the second dose of cefuroxime. Nevertheless, it was reduced after addition of tobramycin. No binding of tobramycin to endotoxin was observed, either in vivo or in vitro. In a porcine sepsis model, a possible anti-inflammatory effect of ceftazidime and tobramycin, expressed as late cytokine inhibition, was seen. The protein synthesis inhibitor, clindamycin, released less streptococcal pyrogenic exotoxin A (SpeA) from a group A streptococcus strain than penicillin, and addition of clindamycin to penicillin resulted in less toxin production than penicillin alone. The SpeA production was dependent on the bacterial number at the start of treatment. Higher doses of penicillin also led to less SpeA. The choice of antibiotic class and dose may be important in the severely ill septic patient in whom an additional toxin release could be deleterious. A combination of a β-lactam antibiotic and a protein synthesis inhibitor seems beneficial but further investigations are needed.

Swine barn air is a heterogeneous mixture of dust, bacteria and irritant chemicals including ammonia and hydrogen sulphide. Gram-negative bacteria are commonly found in swine barn air and significantly contribute to pulmonary disease in unprotected swine barn workers, through the endotoxin moiety, lipopolysaccharide (LPS). Toll-like Receptor-4 is the ligand for LPS. It is found on many cell types including monocytes, macrophages, neutrophils, endothelial cells, and to a lesser extent, epithelial cells. The severity and outcome of acute lung injury following barn air exposures depends upon the balance between epithelial and vascular endothelial repair mechanisms, including angiogenesis. Vascular Endothelial Growth Factor (VEGF) is an endothelial mitogen produced by mesenchymal and alveolar Type II epithelial cells and by activated bronchial airway epithelial cells. Research investigating the role of cytokines in angiogenesis has shown that close proximity of immune cells and endothelial cells modulates the production of various compounds that regulate vascular function. Given that LPS is the ligand for TLR4 there appeared to be a role for TLR4 in angiogenesis, particularly following endotoxin exposure. To determine whether this was occurring, we examined whether exposure to swine barn air alters vascular density in the lungs and the role of TLR4 using a murine model. Toll-like Receptor-4 wild-type (C3HeB/FeJ) and TLR4 mutant (C3H/HeJ) mice were obtained and exposed to swine barn air for 1-, 5-, or 20-days for 8 hours/day. Wild-type animals showed a 127% increase in vascular density after 20-days barn air exposure. Vascular Endothelial Growth Factor-A protein levels were decreased by 0.62-fold after one-day swine barn air exposure in wild-type animals, indicating that VEGF-A is being used as a pro-angiogenic mitogen. Transcription of VEGF-A mRNA was increased in wild-type animals after all swine barn air exposure periods. The receptor VEGFR-1 showed increased mRNA transcription over all time points. These effects were only observed in TLR4 wild-type animals, indicating that these effects are mediated by TLR4. Further, VEGF-A and VEGFR-1 appear to be involved in the manifestation of TLR4-induced angiogenesis in the lung.

Swine barn air is a heterogeneous mixture of dust, bacteria and irritant chemicals including ammonia and hydrogen sulphide. Gram-negative bacteria are commonly found in swine barn air and significantly contribute to pulmonary disease in unprotected swine barn workers, through the endotoxin moiety, lipopolysaccharide (LPS). Toll-like Receptor-4 is the ligand for LPS. It is found on many cell types including monocytes, macrophages, neutrophils, endothelial cells, and to a lesser extent, epithelial cells. The severity and outcome of acute lung injury following barn air exposures depends upon the balance between epithelial and vascular endothelial repair mechanisms, including angiogenesis. Vascular Endothelial Growth Factor (VEGF) is an endothelial mitogen produced by mesenchymal and alveolar Type II epithelial cells and by activated bronchial airway epithelial cells. Research investigating the role of cytokines in angiogenesis has shown that close proximity of immune cells and endothelial cells modulates the production of various compounds that regulate vascular function. Given that LPS is the ligand for TLR4 there appeared to be a role for TLR4 in angiogenesis, particularly following endotoxin exposure. To determine whether this was occurring, we examined whether exposure to swine barn air alters vascular density in the lungs and the role of TLR4 using a murine model. Toll-like Receptor-4 wild-type (C3HeB/FeJ) and TLR4 mutant (C3H/HeJ) mice were obtained and exposed to swine barn air for 1-, 5-, or 20-days for 8 hours/day. Wild-type animals showed a 127% increase in vascular density after 20-days barn air exposure. Vascular Endothelial Growth Factor-A protein levels were decreased by 0.62-fold after one-day swine barn air exposure in wild-type animals, indicating that VEGF-A is being used as a pro-angiogenic mitogen. Transcription of VEGF-A mRNA was increased in wild-type animals after all swine barn air exposure periods. The receptor VEGFR-1 showed increased mRNA transcription over all time points. These effects were only observed in TLR4 wild-type animals, indicating that these effects are mediated by TLR4. Further, VEGF-A and VEGFR-1 appear to be involved in the manifestation of TLR4-induced angiogenesis in the lung.

No description available.

Patienten mit chronischem Nierenversagen leiden sowohl klinisch als auch subklinisch unter Entzündungsepisoden. Um einen Frühindikator der Mikroinflammation zu finden, wurde die Expression funktioneller monozytärer Oberflächenantigene (HLA-DR, CD14, CD16, TLR2 (extra-, intrazellulär), TLR4 (extra-, intrazellulär), CD80, CD86), das Zytokinexpressionsprofil (IL1, IL6, IL10, TNFa, TGFb) und der ultrastruktrurelle Phänotyp des Monozyten in vivo bzw. in vitro untersucht. Dabei wurde strikt zwischen der Membranproteinexpression auf antiinflammatorischen (CD14++CD16-), proinflammatorischen (CD14++CD16+, CD14dimCD16+) und CD14+ bzw. CD16+ Monozyten unterschieden und parallel die Serumspiegel von Parathormon (PTH), C-reaktivem Protein (CRP), Calcium und Phosphor untersucht. Inwieweit monozytäre Aktivierungsmarker zum immunologischen Monitoring geeignet sind, sollte vergleichend zwischen Gesunden, Hämodialyse-(HD)- und Peritonealdialyse-(CAPD)-Patienten untersucht werden. Zusätzlich wurde die Expression in einem in-vitro-Zellkulturmodell vergleichend betrachtet. Der Serum-PTH-Spiegel fiel nach Injektion des Vitamin D-Derivates Paricalcitol, der Serumcalciumspiegel stieg signifikant innerhalb des oberen Referenzbereiches bei Patienten mit sekundärem Hyperparathyroidismus drei Wochen nach Beginn der Therapie. Die HLA-DR, extrazelluläre TLR2, intrazelluläre TLR4, CD80, CD86 Expression fiel nach Paricalcitolinjektion. Paricalcitol erhöhte die Anzahl antiinflammatorischer und erniedrigte die Anzahl proinflammatorischer Monozyten. Beim ultrastrukturellen Vergleich zeigte sich eine deutliche Häufung von elektronendichten Granula bei Paricalcitol-inkubierten Zellen im in-vitro-Versuch. Hierbei handelt es sich mit an Sicherheit grenzender Wahrscheinlichkeit um Lysosomen, was die These einer erhöhten phagolysosomalen monozytären Aktivität unter Paricalcitol-Inkubation stützt. Sowohl bei 48- als auch bei 72-stündiger Inkubation in Primärzellkultur wirkte Paricalcitol antiinflammatorisch, indem es Aktivierungsmarker des Monozyten (HLA-DR, TLR2 (extrazellulär), TLR4 (extrazellulär)), die Anzahl proinflammatorischer Monozyten und die Synthese proinflammatorischer Zytokine (IL1, IL6, TNFa) supprimierte. Dialysepatienten unterliegen einem erhöhten Eintrag von Endotoxin (=LPS) über die Dialysemembran, das in der Regel durch Bindung an den LPS-Rezeptor (=CD14) detoxifiziert wird. Dieser wird nicht nur nach LPS Inkubation, sondern v.a. nach Paricalcitol Inkubation vermehrt exprimiert. LPS erhöhte die Anzahl der proinflammatorischen Monozyten in Zellkultur und reflektierte damit den steady state des HD-Patienten. Aktivierungsmarker von Monozyten unterschieden sich zudem bei vergleichender Betrachtung zwischen CAPD-, HD-Patienten und Gesunden und außerdem im Vergleich vor und nach HD. Die intrazelluläre TLR2 und TLR4 Expression von CAPD-Patienten und HD-Patienten war gegenüber Gesunden erniedrigt, während die CD14 Expression signifikant erhöht war. HD-Patienten zeigten einen erhöhten Anteil proinflammatorischer Monozyten vergleichend zu Gesunden aber auch zu Patienten unter CAPD-Substitutionstherapie. Unmittelbar nach HD fielen die proinflammatorischen Monozyten, während v.a. die Expression von extrazellulärem TLR2, intrazellulärem TLR2 und intrazellulärem TLR4 stieg. Somit unterliegt der HD-Patient einer stärkeren Mikroinflammation als der CAPD-Patient. CAPD-, HD- und auch Patienten mit sekundärem Hyperparathyroidismus zeigen Zeichen einer Mikroinflammation. Dabei war CRP (derzeitiger Routineparameter) kein probates diagnostisches Mittel der Entzündung bei CAPD- und HD-Patienten, des Weiteren nicht geeignet zwischen beiden Patientengruppen zu unterscheiden. Die Verteilung der monozytären Subpopulationen und Expression monozytärer Aktivierungsmarker unterschied sich hinreichend zwischen CAPD- und HD-Patienten. Paricalcitol moduliert funktionelle monozytäre Antigene und Zytokine in vivo und in vitro und wirkt damit der Mikroinflammation und dem Immundefekt des CNI-Patienten entgegen. Der Monozyt wirkt ambivalent und initiiert seine „eigene“ Gegenregulation zur Inflammation, die beim Patienten unter Nierenersatztherapie Endotoxin-vermittelt ist. Vitamin D-Derivate wie Paricalcitol wirken nicht nur auf die Calcium-Phosphat-Homöostase sondern auch immunmodulatorisch, indem sie auf monozytäre Antigene, wie Rezeptoren der angeborenen Immunabwehr, Einfluss nehmen.

Nitric oxide (NO) is an important regulator of pulmonary blood flow and attenuates hypoxic pulmonary vasoconstriction (HPV). Nitric oxide is synthesized enzymatically in a number of tissues, including the lungs, and can also be generated from reduction of nitrite during hypoxia and acidosis. Inhaled nitric oxide (INO) is a selective pulmonary vasodilator, with no effects on systemic arterial blood pressure due to inactivation by hemoglobin in the blood. INO has distant effects both within the lungs and in other organs, since NO can be transported to remote tissues bound to proteins, or as more stable molecules of nitrite and nitrate. In healthy pigs, INO causes vasoconstriction and down regulation of endogenous NO production in lung regions not reached by INO, and predominantly so in hypoxic lung regions, i.e. augmentation of HPV. In this thesis, distant effects of INO in pigs with endotoxemic- and lavage-induced lung injuries were studied. INO increased the NO production in lung regions not reached by INO in endotoxemic pigs, whereas endogenous NO production was unaffected in pigs with lavage-induced injury. Metabolic and/or hypercapnic acidosis frequently occurs in critically ill patients, but whether acidosis affects the endogenous pulmonary NO production is unclear. The regional NO production and blood flow in hyperoxic and hypoxic lung regions, were studied during metabolic and hypercapnic acidosis. Neither metabolic, nor hypercapnic acidosis changed the endogenous NO production in hyperoxic or hypoxic lung regions. Metabolic acidosis potentiated HPV, whereas hypercapnic acidosis transiently attenuated HPV. In conclusion, the present thesis has demonstrated that INO in experimental sepsis increases the endogenous NO production in lung regions not reached by INO, which may cause increased shunt and poor response to INO. This distant effect is not seen in lavage injuried lungs, an experimental model with less inflammation. Acidosis does not affect the endogenous pulmonary NO production in hyperoxic or hypoxic lung regions. Whereas metabolic acidosis potentiates HPV, hypercapnic acidosis transiently attenuates HPV, due to a combination of hypercapnia-induced increase in cardiac output and a probable vasodilating effect of the CO2-molecule.

Made available in DSpace on 2014-06-11T19:30:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-10-17Bitstream added on 2014-06-13T20:21:06Z : No. of bitstreams: 1 rodrigues_la_dr_arafcf.pdf: 878561 bytes, checksum: 1b0302d7b78c8400814eb99c68c33070 (MD5) / Universidade Estadual Paulista (UNESP) / O choque séptico, que é a maior causa de morte no Brasil e no Mundo, consequência de trauma infeccioso, a indução da óxido nítrico sintase (NOS) do endotélio vascular por produtos originados da parede de bactérias é considerado a causa, mas também parte da ativação de mecanismos de defesa do hospedeiro contra a infecção. Uma pronunciada hipotensão seguida de redução da reatividade vascular a mediadores vasoconstritores é determinada pela intensa liberação de mediadores relaxantes, como o próprio óxido nítrico (NO), que podem progredir para a morte do hospedeiro. Em ratos, este fenômeno é igualmente observado. O quadro de choque séptico ou Sepsis pode ser reproduzido pela injeção parenteral de endotoxina de várias bactérias Gram-negativas, como a endotoxina de E. coli (Etx), que produz intensa hipotensão aguda e prolongada, com pronunciada hiporreatividade a angiotensina II (AII)... / In septic schock that is major cause of death folowing in infeccious trauma, the induction of nitric oxide sinthase (NOS) of vascular endothelium by bacterial products is considered to be part of the defense mechanism of hosp against infection. But the hypotension and vascular hyporeactivity determinated by intense release of relaxing factors essentially from endothelial cells, in response to substances, released from walls bcteria which can progress to death of hosp. The hyporreactivity to angiotensin II (AII) is observed in Etx-induced hypotension. The current study show that the vasoconstriction response of AII in rats is reversed by an inhibitor of NOS (N NLA) in control rats, but in adrenalectomized rats these inhibitor did not reversed the hyporreactivity. The dexamethasone impaired the protective effect of N NLA against Etx-induced hyporreactivity . In adrenalectomized rats the N NLA not reverts vascular hyporreactivity to AII. The involvement of antiinflammatory nonsteroids drugs (diclofenac [Diclo] and nimesulide [Nime]) not was involved in reversion of vascular hyporreactivity to AII. Dextran injection, produced hypotension but not produced vascular hyporreactivity to AII. These mechanism is not clarified...(Complete abstract, access undermentioned eletronic address)

Made available in DSpace on 2016-06-02T19:22:12Z (GMT). No. of bitstreams: 1 6524.pdf: 836310 bytes, checksum: 7abc8c7790ee59b230df28eb660ac2b7 (MD5) Previous issue date: 2009-12-17 / Universidade Federal de Sao Carlos / Lipopolysaccharide (LPS), an endotoxin derived from gram-negative bacteria wall, triggers a conjunct of systemic and behavioral responses grouped under the name of sickness behavior. Several works show the inhibition of ingestive behavior in animals with activated immune system by systemic LPS. The aim of this work was demonstrate that LPS inhibit sodium appetite in lower doses than those that cause lethal effects and that LPS inhibitory action on sodium appetite involves α2 adrenergic mechanisms. Were produced three works where we show the LPS effects on sodium appetite, on arterial pressure and in different kinds of thirst. In the first work, we showed that LPS inhibited sodium appetite and only higher doses produced thirst inhibition, besides reduced urinary volume and natriuresis, without alterations on cardiovascular parameters in FURO/CAP-treated rats. LPS abolished intracellular thirst e reduced urinary volume and natriuresis in 2M NaCl gavage-treated animals. LPS also reduced thirst in FURO-treated animals. These results suggest that LPS has a preferential effect on sodium appetite and on thirst intracellular thirst. In the second work we showed that LPS reduce sodium appetite and that this inhibition is abolished by previous treatment with yohimbine ip, an α2 adrenoceptor antagonist in sodium depleted rats. Yohimbine when combined with LPS produced hypotension in sodium depleted rats. LPS reduced gastric empty in sodium depleted rats and this effect wasn‟t abolished by previous treatment with yohimbine. The systemic effects do not explain the inhibition on sodium appetite and we can suggest that LPS exert an inhibitory effect on sodium appetite probably mediated by central α2 adrenoceptors. In the third work we confirmed that LPS inhibit sodium appetite and that central α2 adrenergic ways mediated this response because RX-821002, an α2 adrenoceptor antagonist injected icv, abolished sodium appetite inhibition induced by systemic LPS in sodium depleted rats without produced alterations in cardiovascular parameters. All results exposed with these three works suggest that LPS inhibit sodium appetite and this inhibition is mediated by central α2 adrenoceptors. / Lipopolissacarídeo (LPS), uma endotoxina derivada da parede da bactéria gram-negativa, desencadeia um conjunto de respostas sistêmicas e comportamentais, que se agrupam sob a denominação de comportamento de doença (sickness behavior). Grande número de trabalhos mostra a inibição de comportamento ingestivo no animal que tem ativação do sistema imune induzido por injeção sistêmica de LPS. O objetivo deste trabalho foi demonstrar que o LPS inibe o apetite ao sódio em doses menores daquelas que causam os efeitos letais desta endotoxina e que a ação inibitória do LPS sobre o apetite ao sódio envolve mecanismos adrenérgicos α2. Foram produzidos três trabalhos onde mostramos os efeitos do LPS no apetite ao sódio, na pressão arterial e em diferentes tipos de sede. No primeiro trabalho mostramos que LPS inibiu o apetite ao sódio e apenas as doses mais altas produziram inibição na sede, além de reduzir volume urinário e natriurese, sem alterar os parâmetros cardiovasculares em ratos tratados com o protocolo FURO/CAP. LPS aboliu sede intracelular e reduziu volume urinário e natriurese em ratos que receberam gavagem de NaCl 2M. A sede também foi reduzida pelo tratamento com LPS em ratos tratados com FURO apenas. Esses resultados sugerem que o LPS tem efeito preferencial sobre o apetite ao sódio e sobre a sede intracelular. No segundo trabalho mostramos que LPS reduz o apetite ao sódio e que essa inibição é abolida se há tratamento prévio com ioimbina ip, um antagonista de receptores adrenérgicos α2 em ratos depletados de sódio. Esse mesmo antagonista quando associado com LPS produz hipotensão em ratos depletados de sódio. LPS diminuiu a taxa de esvaziamento gástrico em animais depletados de sódio e esse efeito não foi abolido pelo prévio tratamento com ioimbina. Devido ao fato desses efeitos sistêmicos do LPS não explicarem a inibição no apetite ao sódio podemos sugerir que o LPS exerce um efeito inibitório no apetite ao sódio provavelmente mediado por receptores adrenérgicos α2 centrais. No terceiro trabalho confirmamos que o LPS inibe o apetite ao sódio e que vias centrais noradrenérgicas α2 centrais medeiam essa inibição pois o RX-821002, um antagonista de receptores adrenérgicos α2, injetado icv, também aboliu a inibição no apetite ao sódio induzida pelo LPS sistêmico em ratos depletados de sódio sem produzir alterações nos parâmetros cardiovasculares. Esse conjunto de resultados expostos por meio destes três trabalhos sugere que o LPS inibe o apetite ao sódio e essa inibição é mediada por receptores adrenérgicos α2 de origem central.

Resumo: O choque séptico, que é a maior causa de morte no Brasil e no Mundo, consequência de trauma infeccioso, a indução da óxido nítrico sintase (NOS) do endotélio vascular por produtos originados da parede de bactérias é considerado a causa, mas também parte da ativação de mecanismos de defesa do hospedeiro contra a infecção. Uma pronunciada hipotensão seguida de redução da reatividade vascular a mediadores vasoconstritores é determinada pela intensa liberação de mediadores relaxantes, como o próprio óxido nítrico (NO), que podem progredir para a morte do hospedeiro. Em ratos, este fenômeno é igualmente observado. O quadro de choque séptico ou Sepsis pode ser reproduzido pela injeção parenteral de endotoxina de várias bactérias Gram-negativas, como a endotoxina de E. coli (Etx), que produz intensa hipotensão aguda e prolongada, com pronunciada hiporreatividade a angiotensina II (AII)...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: In septic schock that is major cause of death folowing in infeccious trauma, the induction of nitric oxide sinthase (NOS) of vascular endothelium by bacterial products is considered to be part of the defense mechanism of hosp against infection. But the hypotension and vascular hyporeactivity determinated by intense release of relaxing factors essentially from endothelial cells, in response to substances, released from walls bcteria which can progress to death of hosp. The hyporreactivity to angiotensin II (AII) is observed in Etx-induced hypotension. The current study show that the vasoconstriction response of AII in rats is reversed by an inhibitor of NOS (N NLA) in control rats, but in adrenalectomized rats these inhibitor did not reversed the hyporreactivity. The dexamethasone impaired the protective effect of N NLA against Etx-induced hyporreactivity . In adrenalectomized rats the N NLA not reverts vascular hyporreactivity to AII. The involvement of antiinflammatory nonsteroids drugs (diclofenac [Diclo] and nimesulide [Nime]) not was involved in reversion of vascular hyporreactivity to AII. Dextran injection, produced hypotension but not produced vascular hyporreactivity to AII. These mechanism is not clarified...(Complete abstract, access undermentioned eletronic address) / Orientador: Maria Teresa Pepato / Coorientador: José Francisco Fracasso / Banca: José Ranali / Banca: Neoclair Molina / Banca: Wilson Abrão Saad / Banca: Iguatemy Lourenço Brunetti / Doutor

O objetivo deste trabalho foi avaliar a capacidade da solução de nanopartículas de prata, utilizada como irrigante e medicação intracanal, em controlar os microrganismos e neutralizar endotoxinas no canal radicular. Para isso, foram utilizadas 48 raízes de dentes humanos padronizadas em 16 mm e com diâmetro apical correspondente a uma lima tipo Kerr no. 30. Os canais foram contaminados por 28 dias com E. coli e por 21 dias com E. faecalis e C. albicans. Os espécimes foram divididos em quatro grupos (n=12), de acordo com a substância utilizada (solução irrigadora e medicação intracanal): G1) solução salina e solução salina (grupo controle); G2) hipoclorito de sódio 1% associado ao creme Endo-PTC e hidróxido de cálcio - protocolo tradicional da FOUSP; G3) solução de nanopartículas de prata 50 ppm e hidróxido de cálcio associado à solução de nanopartículas de prata; e G4) solução de nanopartículas de prata 50 ppm e solução de nanopartículas de prata 50 ppm. Foram realizadas cinco coletas do conteúdo do canal radicular para avaliar a atividade antimicrobiana: coleta de confirmação, imediatamente após a instrumentação (1ª. coleta) e outra após sete dias (2ª. coleta); imediatamente após a remoção da medicação (3ª. coleta) e outra após sete dias (4ª. coleta). A neutralização da endotoxina foi avaliada apenas nas quatro últimas coletas. Os resultados obtidos foram submetidos a análise estatística (Kruskall-Wallis e Dunn). Todas as soluções irrigadoras promoveram redução significativa dos microrganismos após a instrumentação (1ª. coleta) (p<0,05). Após 7 dias houve aumento do número de microrganismos em todos os grupos, voltando ao número inicial nos grupos instrumentados com solução salina e hipoclorito de sódio. Entretanto, apesar de ter ocorrido recolonização dos microrganismos no canal radicular dos espécimes instrumentados com solução de nanopartículas de prata, manteve-se redução estatisticamente significantemente do número de UFC/mL quando comparada à coleta de confirmação (p<0,05), demonstrando efeito residual. A medicação de hidróxido de cálcio eliminou 100% dos microrganismos e manteve os resultados após 7 dias (4ª. coleta) no grupo 2. No grupo quatro houve recolonização de E. faecalis e C. albicans após 7 dias de incubação. O hidróxido de cálcio foi a única substância avaliada que promoveu redução significativa (p<0,05) das endotoxinas. Assim, pôde-se concluir que o hipoclorito de sódio e a solução de NP-Ag reduziram significativamente a microbiota do canal, no entanto somente com a associação ao hidróxido de cálcio houve eliminação dos microrganismos em profundidade nos túbulos dentinários e redução das endotoxinas presentes no canal radicular. / The objective of this study was to evaluate the antimicrobial effect and endotoxin detoxified of solution of silver nanoparticles used as irrigating and dressing in root canals. Forty-eigth single-root human teeth were used. All root canals sized 16 mm and were enlarged to a Kerr file number .30. Root canals were infected for 28 days with E. coli and for 21 days with E. faecalis and C. albicans. The specimens were divided into four groups (n=12), according to the substance used (irrigating solution and dressing): G1) saline and saline (control), G2) sodium hypochlorite 1% with Endo-PTC cream and calcium hydroxide; G3) solution of Ag-NP 50 ppm and calcium hydroxide with Ag-NP; and G4) solution of Ag-NP 50 ppm and solution of Ag-NP 50 ppm. Five samplings of root canal were accomplished to evaluate the antimicrobial activity: confirmation sample, immediately after instrumentation (1st. Sample), and other after 7 days (2 nd. Sample), immediately after dressing removed (3 rd. Sample) and other after 7 days (4 th. Sample). The endotoxina detoxified was evaluated only in the last four samples by Limulus assay. Results were analysed by Kruskal-Wallis and Dunn. All irrigation solutions caused significant reduction of microorganisms after instrumentation (1 st. Collection) (p <0.05). After 7 days there was increase of the number of bacteria in all groups, returning to the initial number of microorganisms in the groups prepared with saline and sodium hypochlorite, however, the recolonization of microorganisms in root canals of groups of solution silver nanoparticles was smaller than the number of confirmation sample (p <0.05), showing residual effect. The calcium hydroxide eliminated 100% of the microorganisms and the results remained after 7 days (4 th. Collection) for group 2. In group 4 there was recolonization of E. faecalis and C. albicans after 7 days. Calcium hydroxide was the only substance that measured a significant reduction of endotoxin (p <0.05). Thus, we concluded that the sodium hypochlorite and the NP-Ag significantly reduced the microorganisms of the root canal, but it is necessary the combination of calcium hydroxide to eliminated all microorganisms into dentinal tubules and detoxified endotoxins in the root canal.