RESUMO
PURPOSE: To assess the distribution of microorganisms isolated from patients with bacterial endophthalmitis and their antimicrobial susceptibility. METHODS: Retrospective analysis of medical and microbiological records of patients with suspected diagnosis of endophthalmitis and bacterial culture-proven at the Department of Ophthalmology, UNIFESP, between January 1 2000 and December 31 2005. RESULTS: 153 (33.9%) of 451 patients showed positive bacterial culture. A total of 155 microorganisms were isolated, 79.35% were gram-positive and 20.65% gram-negative. Staphylococcus (CoNS) (41.94%) were the most frequently isolated. The antimicrobial susceptibility for gram-negative microorganisms was as follows: amikacin 87.10%, tobramycin 80.65%, ciprofloxacin 96.67%, levofloxacin, gatifloxacin and moxifloxacin 100%, ceftazidime 85.0%, and gentamicin 80.65%. Vancomycin sensitivity among gram-positive microorganisms was 100%. S. aureus and CoNS showed 83.33% of susceptibility to oxacillin, 89.61% to ciprofloxacin and 100% to gatifloxacin and moxifloxacin. The main acquisition mechanism was postoperative (60.65%). CONCLUSION: We detected a low sensitivity of vitreous/aqueous culture for the etiologic diagnosis of endophthalmitis. The empiric antimicrobial therapy or prophylaxis should be active against gram-positive bacteria, particularly staphylococci. Surveillance studies of bacterial resistance are important for a better utilization of antimicrobials in this clinical setting.
Assuntos
Antibacterianos/farmacologia , Endoftalmite/microbiologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Antibacterianos/uso terapêutico , Endoftalmite/tratamento farmacológico , Bactérias Gram-Negativas/classificação , Bactérias Gram-Negativas/isolamento & purificação , Bactérias Gram-Positivas/classificação , Bactérias Gram-Positivas/isolamento & purificação , Humanos , Testes de Sensibilidade Microbiana , Estudos RetrospectivosRESUMO
PURPOSE: To determine the efficacy of freeze-dried amniotic membrane (AM) for reconstruction of the ocular surface in rabbit eyes. METHODS: The sterilized, freeze-dried amniotic membrane (lyophilized or FD-AM) is a preservative method that uses the drying by freezing process to maintain the AM well preserved for a long time even at room temperature. This paper is an experimental animal interventional study. One eye of each of 15 male New Zealand rabbits (1.5 - 3.0 kg) had the central cornea marked with a 6.0 mm trephine. The marked area was deepithelialized with a No.15 blade. The denuded corneal surface was covered as follows: Group 1: cryopreserved AM (n=6); Group 2: freeze-dried AM (n=6); and Group 3: not covered (control group, n=3). The AM in group 1 and 2 and the periphery of the denuded area in group 3 were secured with continuous 10-0 nylon sutures. The clinical evaluation was made by a blinded observer and graded on a four-point scale (1= minimal, 4= marked) for conjunctival and ciliary hyperemia, eyelid edema, corneal neovascularization, corneal opacity and reepithelialization on postoperative (PO) days 1, 7 and 30 . After PO day 30, the rabbits were euthanized and their corneas were sent for histopathological and ultrastructural analysis to evaluate tissue inflammation, reepithelialization, and basement membrane integrity. RESULTS: Two eyes in group 2 had a corneal infection and were excluded from the analysis. No statistically significant differences among the three groups were found (p>0.05) regarding the clinical evaluation on 1st, 7th and 30th PO days. On transmission electron microscopy, the basement membrane in lyophilized and control groups was more continuous and homogeneous than in the glycerol group. CONCLUSIONS: The freeze-drying method seems to be a good option to preserve human amniotic membrane to be used in ocular surface reconstruction. This preservative method reduces the preservation costs and may enhance the use of AM, facilitating its storage and transport.
Assuntos
Âmnio/transplante , Córnea/cirurgia , Criopreservação/métodos , Âmnio/ultraestrutura , Animais , Córnea/ultraestrutura , Liofilização , Masculino , CoelhosRESUMO
OBJETIVO: Determinar a distribuição dos microrganismos isolados de pacientes com endoftalmite bacteriana e sua sensibilidade a antimicrobianos. MÉTODOS: Foram analisados retrospectivamente os dados clínicos e microbiológicos dos pacientes com hipótese diagnóstica de endoftalmite e cultura bacteriana positiva, atendidos no Departamento de Oftalmologia da UNIFESP de 1º de janeiro de 2000 a 31 de dezembro de 2005. RESULTADOS: De 451 pacientes, 153 (33,9 por cento) apresentaram cultura bacteriana positiva. Foram isolados 155 microrganismos, sendo 79,35 por cento gram-positivos e 20,65 por cento gram-negativos. Os Staphylococcus coagulase-negativos (SCoN) (41,94 por cento) foram os mais freqüentemente isolados. A sensibilidade aos antimicrobianos entre os gram-negativos foi: amicacina 87,10 por cento, tobramicina 80,65 por cento, ciprofloxacina 96,67 por cento, levofloxacina, gatifloxacina e moxifloxacina 100 por cento, ceftazidima 85 por cento, e gentamicina 80,65 por cento. A sensibilidade à vancomicina entre os gram-positivos foi de 100 por cento. S. aureus e SCoN apresentaram 83,33 por cento de sensibilidade à oxacilina, 89,61 por cento à ciprofloxacina e 100 por cento à gatifloxacina e moxifloxacina. A forma de aquisição predominante foi a pós-operatória (60,65 por cento). CONCLUSÃO: Observamos baixa sensibilidade da cultura para o diagnóstico etiológico das endoftalmites. Uma terapia antimicrobiana ou profilaxia empírica deve ser ativa contra os microrganismos gram-positivos, particularmente contra estafilococos. Estudos de vigilância de resistência bacteriana são importantes para adequação desses esquemas.
PURPOSE: To assess the distribution of microorganisms isolated from patients with bacterial endophthalmitis and their antimicrobial susceptibility. METHODS: Retrospective analysis of medical and microbiological records of patients with suspected diagnosis of endophthalmitis and bacterial culture-proven at the Department of Ophthalmology, UNIFESP, between January 1 2000 and December 31 2005. RESULTS: 153 (33.9 percent) of 451 patients showed positive bacterial culture. A total of 155 microorganisms were isolated, 79.35 percent were gram-positive and 20.65 percent gram-negative. Staphylococcus (CoNS) (41.94 percent) were the most frequently isolated. The antimicrobial susceptibility for gram-negative microorganisms was as follows: amikacin 87.10 percent, tobramycin 80.65 percent, ciprofloxacin 96.67 percent, levofloxacin, gatifloxacin and moxifloxacin 100 percent, ceftazidime 85.0 percent, and gentamicin 80.65 percent. Vancomycin sensitivity among gram-positive microorganisms was 100 percent. S. aureus and CoNS showed 83.33 percent of susceptibility to oxacillin, 89.61 percent to ciprofloxacin and 100 percent to gatifloxacin and moxifloxacin. The main acquisition mechanism was postoperative (60.65 percent). CONCLUSION: We detected a low sensitivity of vitreous/aqueous culture for the etiologic diagnosis of endophthalmitis. The empiric antimicrobial therapy or prophylaxis should be active against gram-positive bacteria, particularly staphylococci. Surveillance studies of bacterial resistance are important for a better utilization of antimicrobials in this clinical setting.
Assuntos
Humanos , Antibacterianos/farmacologia , Endoftalmite/microbiologia , Bactérias Gram-Negativas/efeitos dos fármacos , Bactérias Gram-Positivas/efeitos dos fármacos , Antibacterianos/uso terapêutico , Endoftalmite/tratamento farmacológico , Bactérias Gram-Negativas/classificação , Bactérias Gram-Negativas/isolamento & purificação , Bactérias Gram-Positivas/classificação , Bactérias Gram-Positivas/isolamento & purificação , Testes de Sensibilidade Microbiana , Estudos RetrospectivosRESUMO
PURPOSE: To determine the efficacy of freeze-dried amniotic membrane (AM) for reconstruction of the ocular surface in rabbit eyes. METHODS: The sterilized, freeze-dried amniotic membrane (lyophilized or FD-AM) is a preservative method that uses the drying by freezing process to maintain the AM well preserved for a long time even at room temperature. This paper is an experimental animal interventional study. One eye of each of 15 male New Zealand rabbits (1.5 - 3.0 kg) had the central cornea marked with a 6.0 mm trephine. The marked area was deepithelialized with a No.15 blade. The denuded corneal surface was covered as follows: Group 1: cryopreserved AM (n=6); Group 2: freeze-dried AM (n=6); and Group 3: not covered (control group, n=3). The AM in group 1 and 2 and the periphery of the denuded area in group 3 were secured with continuous 10-0 nylon sutures. The clinical evaluation was made by a blinded observer and graded on a four-point scale (1= minimal, 4= marked) for conjunctival and ciliary hyperemia, eyelid edema, corneal neovascularization, corneal opacity and reepithelialization on postoperative (PO) days 1, 7 and 30 . After PO day 30, the rabbits were euthanized and their corneas were sent for histopathological and ultrastructural analysis to evaluate tissue inflammation, reepithelialization, and basement membrane integrity. RESULTS: Two eyes in group 2 had a corneal infection and were excluded from the analysis. No statistically significant differences among the three groups were found (p>0.05) regarding the clinical evaluation on 1st, 7th and 30th PO days. On transmission electron microscopy, the basement membrane in lyophilized and control groups was more continuous and homogeneous than in the glycerol group. CONCLUSIONS: The freeze-drying method seems to be a good option to preserve human amniotic membrane to be used in ocular surface reconstruction. This preservative method reduces the preservation costs and may enhance...
OBJETIVO: Avaliar a eficácia da liofilização da membrana amniótica (MA) para a reconstrução da superfície ocular em coelhos. MÉTODOS: A liofilização é processo de preservação que mantém a MA estável durante longo tempo mesmo em temperatura ambiente. A córnea de um olho de cada coelho macho da raça Nova Zelândia foi marcada e desepitelizada. Essa área desepitelizada foi coberta com: Grupo 1: MA criopreservada (n=6); Grupo 2: MA liofilizada (n=6) e Grupo 3: Não coberta (n=3). A MA nos grupos 1 e 2 e a periferia da córnea no grupo 3 foram suturadas com nylon 10-0. A avaliação clínica foi realizada por um observador cego em relação à hiperemia, neovascularização e edema de córnea e reepitelização nos dia 1, 7 e 30 pós-operatórios. Após o dia 30 os ratos foram eutanizados e suas córneas enviadas para análise histopatológica e ultra-estrutural. RESULTADOS: Dois olhos no grupo 2 foram excluídos da análise devido à infecção. Não foi encontrada diferença estatisticamente significante entre os grupos em relação à avaliação clínica. Na microscopia eletrônica de transmissão, a membrana basal nos grupos de MA liofilizada e controle foi mais contínua e homogênea em relação ao grupo da MA criopreservada. CONCLUSÕES: O processo de liofilização parece ser boa opção para a preservação da membrana amniótica humana para utilização na reconstrução da superfície ocular.
Assuntos
Animais , Masculino , Coelhos , Âmnio/transplante , Córnea/cirurgia , Criopreservação/métodos , Âmnio/ultraestrutura , Córnea/ultraestrutura , LiofilizaçãoRESUMO
PURPOSE: To evaluate the epidemiology of superficial corneal foreign body. METHODS: Patients who were seen at the Emergency Service of the Federal University of São Paulo, from April/05 to June/05, were screened and those with superficial corneal foreign body were interviewed. Data regarding gender, age, occupation, employment status, availability and use of protective devices and supervision of their use were collected. Awareness of the possible complications was also assessed. Results were analyzed using chi-square or Fisher exact test. RESULTS: One hundred twenty-three patients were interviewed. Only 3 patients were female. The mean age was 36 years. Most injuries occurred at the workplace (86.2%), and 58.4% of the patients did not have a legal employment registration. The occupational activities most frequently reported were construction related activities (44.3%) and welding/soldering (11.3%). In most workplaces (79.8%) protective devices were available and 85.3% of the patients were instructed to use them. A safety device was being used during the accident in 34.2% of the cases and this was more frequent among patients that had legal employment registry (p=0.008) and among those under supervision (p=0.0415). The majority of the patients (68.9%) were aware of the risk of severe complications. CONCLUSIONS: Most patients with superficial corneal foreign body are aware of its severe complications and injuries usually occur in places where safety devices are available and often during their use. Our findings suggest that prevention should focus on supervision and correct use of safety devices.
Assuntos
Acidentes de Trabalho/estatística & dados numéricos , Corpos Estranhos no Olho/epidemiologia , Ocupações , Soldagem , Acidentes de Trabalho/prevenção & controle , Adolescente , Adulto , Idoso , Conscientização , Brasil/epidemiologia , Distribuição de Qui-Quadrado , Materiais de Construção/efeitos adversos , Emprego , Corpos Estranhos no Olho/complicações , Corpos Estranhos no Olho/prevenção & controle , Dispositivos de Proteção dos Olhos/estatística & dados numéricos , Feminino , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Masculino , Pessoa de Meia-Idade , Sistema de Registros , Local de TrabalhoRESUMO
OBJETIVO: Avaliar a epidemiologia do trauma ocular por corpo estranho superficial de córnea. MÉTODOS: Os pacientes atendidos no Pronto-Socorro da Universidade Federal de São Paulo entre abril e junho de 2005 que apresentaram corpo estranho superficial de córnea foram entrevistados. Foram avaliados: sexo, idade, profissão, registro legal do emprego, uso, disponibilidade e tipo de equipamentos de proteção utilizados e a fiscalização do seu uso. O conhecimento das complicações deste tipo de acidente também foi avaliado. Os resultados foram analisados com teste do qui quadrado ou teste de Fisher. RESULTADOS: Foram entrevistados 123 pacientes. Apenas 3 eram do sexo feminino e a idade média foi de 36 anos. A maioria destes traumas ocorreu no ambiente de trabalho (86,2 por cento) e 58,4 por cento não possuíam registro legal do emprego. As profissões mais freqüentemente envolvidas foram serralheiro, pedreiro e metalúrgico. Em 79,8 por cento dos locais de trabalho havia equipamentos de proteção e 85,3 por cento dos pacientes eram orientados a usá-los. Em 52,4 por cento dos locais sua utilização era fiscalizada, mas apenas 34,2 por cento usavam no momento do trauma. A utilização foi mais freqüente (p=0,008) e fiscalização mais presente (p=0,0415) entre pacientes com registro legal de emprego. Questionados sobre os riscos, 68,9 por cento dos pacientes tinham consciência das complicações graves deste tipo de acidente. CONCLUSÃO: A maioria dos pacientes tem conhecimento sobre a gravidade do trauma ocular e este tipo de lesão ocorre mesmo em locais com equipamentos de proteção disponíveis, alguns deles até durante o seu uso. Os dados sugerem que enfoque maior da prevenção deve ser na fiscalização e utilização de equipamentos adequados.
PURPOSE: To evaluate the epidemiology of superficial corneal foreign body. METHODS: Patients who were seen at the Emergency Service of the Federal University of São Paulo, from April/05 to June/05, were screened and those with superficial corneal foreign body were interviewed. Data regarding gender, age, occupation, employment status, availability and use of protective devices and supervision of their use were collected. Awareness of the possible complications was also assessed. Results were analyzed using chi-square or Fisher exact test. RESULTS: One hundred twenty-three patients were interviewed. Only 3 patients were female. The mean age was 36 years. Most injuries occurred at the workplace (86.2 percent), and 58.4 percent of the patients did not have a legal employment registration. The occupational activities most frequently reported were construction related activities (44.3 percent) and welding/soldering (11.3 percent). In most workplaces (79.8 percent) protective devices were available and 85.3 percent of the patients were instructed to use them. A safety device was being used during the accident in 34.2 percent of the cases and this was more frequent among patients that had legal employment registry (p=0.008) and among those under supervision (p=0.0415). The majority of the patients (68.9 percent) were aware of the risk of severe complications. CONCLUSIONS: Most patients with superficial corneal foreign body are aware of its severe complications and injuries usually occur in places where safety devices are available and often during their use. Our findings suggest that prevention should focus on supervision and correct use of safety devices.
Assuntos
Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Acidentes de Trabalho/estatística & dados numéricos , Corpos Estranhos no Olho/epidemiologia , Ocupações , Soldagem , Conscientização , Acidentes de Trabalho/prevenção & controle , Brasil/epidemiologia , Distribuição de Qui-Quadrado , Materiais de Construção/efeitos adversos , Emprego , Corpos Estranhos no Olho/complicações , Corpos Estranhos no Olho/prevenção & controle , Dispositivos de Proteção dos Olhos , Conhecimentos, Atitudes e Prática em Saúde , Sistema de Registros , Local de TrabalhoRESUMO
PURPOSE: To compare the anatomical structure and the presence of growth factors and cytokines of amniotic membrane preserved in glycerol/MEM (1:1) or undiluted dimethyl sulfoxide through electron microscopy. METHODS: Amniotic membrane preserved in glycerol/MEM (1:1) or undiluted dimethyl sulfoxide were processed for transmission and scanning electron microscopy. As control, freshly collected amniotic membrane was fixed and processed for electron microscopy. The cytokines and growth factors assessed were: TGF-beta (transforming growth factor beta); TGF-beta activ (activated transforming growth factor beta); EGF (epidermal growth factor); FGF-4 (fibroblast growth factor 4); bFGF (basic fibroblast growth factor); IL-4 (interleukin 4); PGE2 (prostaglandin E2); IL-10 (interleukin 10); KGF (keratinocyte growth factor); HGF (hepatocyte growth factor). RESULTS: Amniotic membrane from the control group showed intact epithelium, with surface microvilli and junctional complexes between the cells and the basal membrane. Glycerol/MEM preserved amniotic membrane had similar aspect to the control, with higher epithelial cells. Those amniotic membranes preserved in dimethyl sulfoxide disclosed less intercellular junction and detachment of the epithelium from the basal membrane. The cytokines and growth factors did not disclose significant differences, except for FGF-4, bFGF, PGE2 and KGF. CONCLUSIONS: Amniotic membrane preserved in glycerol/MEM showed a better tissue structure, with less detachment of the epithelium from the basal membrane, in comparison to undiluted dimethyl sulfoxide. The majority of the growth factors and cytokines were kept with both techniques of preservation.
Assuntos
Âmnio/metabolismo , Âmnio/ultraestrutura , Criopreservação/métodos , Crioprotetores/farmacologia , Dimetil Sulfóxido/farmacologia , Glicerol/farmacologia , Âmnio/efeitos dos fármacos , Proliferação de Células , Células Cultivadas , Citocinas/metabolismo , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/ultraestrutura , Humanos , Microscopia Eletrônica de Transmissão , Fator de Crescimento Transformador beta/metabolismoRESUMO
OBJETIVO: Comparar, por microscopia eletrônica, a integridade anatômica e a presença de fatores de crescimento e citocinas da membrana amniótica preservada com glicerol/MEM (1:1) e dimetilsulfóxido puro. MÉTODOS: As membranas amnióticas preservadas em glicerol/MEM (1:1) ou dimetilsulfóxido puro foram processadas para microscopia eletrônica de transmissão e varredura. Como controle, membrana amniótica fresca foi imediatamente fixada após coleta e processada para microscopia eletrônica. As citocinas e os fatores de crescimento avaliados foram: TGF-beta- fator transformador de crescimento beta; TGF-beta ativ- fator transformador de crescimento beta ativado; EGF- fator recombinante de crescimento epitelial humano; FGF-4- fator de crescimento fibroblástico 4; FGF-beta- fator de crescimento fibroblástico básico; IL-4- interleucina 4; PGE2- prostaglandina E2; IL-10- interleucina 10; KGF- fator de crescimento de queratinócito; HGF- fator de crescimento de hepatócito. RESULTADOS: As membranas amnióticas do grupo controle apresentavam epitélio íntegro, com microvilos na superfície e complexos juncionais entre as células e a membrana basal. As membranas amnióticas preservadas em glicerol/MEM tinham aspecto semelhante às do controle, com maior altura das células epiteliais. Já as membranas amnióticas preservadas em dimetilsulfóxido mostraram redução das junções intercelulares e destacamento do epitélio da membrana basal. As citocinas e fatores de crescimento não apresentaram diferenças entre os grupos, exceto FGF-4, FGF-beta, PGE2 e KGF. CONCLUSÕES: A membrana amniótica preservada em meio glicerol/MEM apresentou melhor integridade tecidual, com menor desprendimento do epitélio da membrana basal, em comparação com a preservada no dimetilsulfóxido puro. Os fatores de crescimento e citocinas estavam, em sua maior parte, preservados com as duas técnicas de preservação.
PURPOSE: To compare the anatomical structure and the presence of growth factors and cytokines of amniotic membrane preserved in glycerol/MEM (1:1) or undiluted dimethyl sulfoxide through electron microscopy. METHODS: Amniotic membrane preserved in glycerol/MEM (1:1) or undiluted dimethyl sulfoxide were processed for transmission and scaning electron microscopy. As control, freshly collected amniotic membrane was fixed and processed for electron microscopy. The cytokines and growth factors assessed were: TGF-beta (transforming growth factor beta); TGF-b activ (activated transforming growth factor beta); EGF (epidermal growth factor); FGF-4 (fibroblast growth factor 4); bFGF (basic fibroblast growth factor); IL-4 (interleukin 4); PGE2 (prostaglandin E2); IL-10 (interleukin 10); KGF (keratinocyte growth factor); HGF (hepatocyte growth factor). RESULTS: Amniotic membrane from the control group showed intact epithelium, with surface microvilli and junctional complexes between the cells and the basal membrane. Glycerol/MEM preserved amniotic membrane had similar aspect to the control, with higher epithelial cells. Those amniotic membranes preserved in dimethyl sulfoxide disclosed less intercellular junction and detachment of the epithelium from the basal membrane. The cytokines and growth factors did not disclose significant differences, except for FGF-4, bFGF, PGE2 and KGF. CONCLUSIONS: Amniotic membrane preserved in glycerol/MEM showed a better tissue structure, with less detachment of the epithelium from the basal membrane, in comparison to undiluted dimethyl sulfoxide. The majority of the growth factors and cytokines were kept with both techniques of preservation.
Assuntos
Humanos , Âmnio/metabolismo , Âmnio/ultraestrutura , Criopreservação/métodos , Crioprotetores/farmacologia , Dimetil Sulfóxido/farmacologia , Glicerol/farmacologia , Âmnio/efeitos dos fármacos , Proliferação de Células , Células Cultivadas , Citocinas/metabolismo , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/ultraestrutura , Microscopia Eletrônica de Transmissão , Fator de Crescimento Transformador beta/metabolismoRESUMO
OBJETIVO: Avaliar as características morfológicas da membrana amniótica desepitelizada por diferentes técnicas. MÉTODOS: A membrana amniótica humana foi coletada no momento do parto, fixada em concentrações crescentes de glicerol (0-50 por cento em DMEM) e preservada a 80°C até a hora de ser usada. O estudo consistiu de 4 grupos: epitélio intacto (controle) e membranas desepitelizadas pela tripsina (2 mg/mL a 1:250), dispase (1,2 U/mL em solução salina balanceada de Hank livre de Mg2+ e Ca2+) e ácido etilenodiaminotetra-acético (EDTA), 0,02 por cento). As amostras foram submetidas à análise por microscopia eletrônica (de varredura e de transmissão). RESULTADOS: A microscopia eletrônica de varredura mostrou epitélio intacto no grupo controle e sua ausência nas membranas amnióticas desepitelizadas pela tripsina e pela dispase. Naquelas tratadas com o ácido etilenodiaminotetra-acético, havia áreas com e sem epitélio. Quando avaliadas pela microscopia eletrônica de transmissão, o epitélio estava intacto e firmemente aderido à membrana basal através de hemidesmossomos nos grupos controle e em parte do ácido etilenodiaminotetra-acético. Havia apenas fibras colágenas nas membranas tratadas com dispase e tripsina. CONCLUSÕES: O tratamento da membrana amniótica com tripsina e dispase pode causar completa retirada do epitélio e da membrana basal, ao passo que o ácido etileno- diaminotetra-acético pode preservar áreas com epitélio intacto e parcialmente destruir a membrana basal em outras.
PURPOSE: To evaluate the morphological features of the amniotic membrane denuded by different techniques. METHODS: Human amniotic membrane was collected at the time of delivery, fixed in increasing concentrations of glycerol (0-50 percent in DMEM) and preserved at -80°C until the time of use. The study consisted of 4 groups: intact epithelium (control) and denuded by trypsin (2 mg/mL at 1:250), dispase (1.2 U/mL in Mg2+ and Ca2+ free Hank's balanced salt solution) or ethylenediaminetetraacetic acid (EDTA), 0.02 percent. Specimens were submitted to electron (scanning and transmission) microscopy analysis. RESULTS: Scanning electron microscopy disclosed intact epithelium in the control group and its absence in the amniotic membranes denuded by trypsin and dispase. In those denuded by ethylenediaminetetraacetic acid there were areas with and without epithelium. When assessed by transmission electron microscopy, the epithelium was intact and firmly adhered to the basement membrane by hemidesmossomes in controls and in parts of ethylenediaminetetraacetic acid group. There were only collagen fibers in the dispase- and trypsin-treated groups. CONCLUSIONS: Trypsin and dispase treatment of the amniotic membrane may cause complete denuding of the epithelium and basement membrane whereas ethylenediaminetetraacetic acid may leave some intact epithelium-areas and partially destroy the basement membrane in others.
Assuntos
Humanos , Âmnio/ultraestrutura , Células Epiteliais/ultraestrutura , Âmnio/efeitos dos fármacos , Técnicas de Cultura de Células , Ácido Edético/farmacologia , Endopeptidases/farmacologia , Microscopia Eletrônica , Tripsina/farmacologiaRESUMO
PURPOSE: The use of medicinal plants for the treatment of human diseases has increased worldwide. Many of them are used by oral administration and, after absorption, may affect many organs. Therefore, this study aimed at assessing the effects of the aqueous extract of Sida cordifolia leaves, popularly known in Brazil as "malva-branca", on liver regeneration. METHODS: Twenty rats were divided into four groups: control, Sida100, Sida200 and Sida400 groups. All animals were submitted to oral administration of distilled water, 100, 200 and 400 mg/kg of the aqueous extract of Sida cordifolia, respectively. Immediately after this, they underwent 67% partial hepatectomy. Twenty four hours later, their livers were removed. Hepatic regeneration was assessed by immunohistochemical staining for proliferating cell nuclear antigen (PCNA) using the PC-10 monoclonal antibody. RESULTS: Sida100 and Sida200 groups disclosed higher liver regeneration indices than control group (p<0.001 and p<0.05, respectively). CONCLUSION: The aqueous extract of Sida cordifolia stimulates liver regeneration after 67% partial hepatectomy in rats.