Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da Universidade Federal do Ceará (UFC) |
Texto Completo: | http://www.repositorio.ufc.br/handle/riufc/56218 |
Resumo: | Gastroesophageal reflux disease (GERD) is a prevalent condition that affects about 13% of the world population. Approximately 60-70% of patients do not present macroscopic changes in the esophagus, while about 20-30% have erosions on endoscopy. Although PPI therapy is the treatment of choice, about a third of patients do not respond adequately to treatment, especially those with Non-Erosive Reflux Disease (NERD). Therefore, alternative forms of treatment should be considered. We aimed to evaluate the integrity of the esophageal mucosa in patients with gastroesophageal reflux disease and to investigate the topical protective effect of polysaccharide Anadenanthera colubrina (Angico Gum, AG in acronym). Six biopsies of the distal esophageal mucosa were collected from 35 patients (24 without erosion or NERD and 11 with erosive esophagitis or EE) and mounted in Ussing chambers. AG was applied topically and in different concentrations (1%, 3% and 10%, w/v), followed by exposure with pH 2.0 acid solution containing pepsin and taurodeoxycholic acid (TDCA) for 30 min, where Transepithelial Electrical Resistance (TER) was continuously measured. Control biopsies (no coated) were also exposed to acid solution. Then, the acid solution was replaced by the fluorescein solution, to assess transepithelial permeability for 90 min. In another experiment, the control and pretreated biopsies with AG at the concentration of 10% were exposed to acid solution pH 2.0 containing pepsin and TDCA for 120 min. The TER of the control biopsies and pretreated with AG-10% or sodium alginate was also evaluated after exposure to acid solution pH 1.0 containing pepsin and TDCA for 30 minutes, and then the acid solution was replaced by the fluorescein solution, for evaluation of the transepithelial permeability for 90 min. We performed an epifluorescence microscopy to localize on the oesopageal epithelium the biopolymer previously labeled with FITC. The pH 2.0 acid solution containing pepsin and TDCA promoted a gradual drop in TER in control biopsies of NERD patients and EE at the end of 30 min Pretreatment with AG-10% was able to prevent the drop in TER (P<0,05) compared to control biopsies. AG-1% and AG-3% did not protect the esophageal mucosa against the refluxate-like solution (P> 0.05). Acid exposure (pH 2.0 + TDCA + pepsin) on the luminal side of the esophageal mucosa of NERD and EE patients promoted a gradual increase in fluorescein permeability, with maximum effect over 90 min. Topical application of AG-10% significantly decreases (P <0.05) the fluorescein permeability in NERD patients, however, this reduction was not significant in EE patients (P> 0.05). The protective effect of AG-10% was sustained for up to 120 min both in biopsies of NERD and EE (P <0.05), when compared to control biopsies. AG-10% and sodium alginate were not able to protect biopsies against more acidic solution (pH 1.0) containing pepsin and TDCA when compared to control biopsies (P> 0.05). AG-10% and alginate were not able to decrease the fluorescein permeability when compared to the control group. Epifluorescent microscope images showed mucosal luminal adherence of FITC- labeled AG-10%. We concluded that AG-10% applied to mucosal biopsies of patients with NERD and EE has a prolonged topical protective effect against exposure to acid solution (pH 2.0 + TDCA + pepsin). However, this effect was not observed with the exposure of biopsies to more acidic solutions (pH 1.0 + TDCA + pepsin). These findings should be further explored in the development of new topical treatments for GERD patients. |
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Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrinaIn vitro topical protection of esophageal mucosa from patients with gerd using “angico gum” a biopolymer from Anadenanthera colubrinaMucosa EsofágicaRefluxo GastroesofágicoGomas VegetaisGastroesophageal reflux disease (GERD) is a prevalent condition that affects about 13% of the world population. Approximately 60-70% of patients do not present macroscopic changes in the esophagus, while about 20-30% have erosions on endoscopy. Although PPI therapy is the treatment of choice, about a third of patients do not respond adequately to treatment, especially those with Non-Erosive Reflux Disease (NERD). Therefore, alternative forms of treatment should be considered. We aimed to evaluate the integrity of the esophageal mucosa in patients with gastroesophageal reflux disease and to investigate the topical protective effect of polysaccharide Anadenanthera colubrina (Angico Gum, AG in acronym). Six biopsies of the distal esophageal mucosa were collected from 35 patients (24 without erosion or NERD and 11 with erosive esophagitis or EE) and mounted in Ussing chambers. AG was applied topically and in different concentrations (1%, 3% and 10%, w/v), followed by exposure with pH 2.0 acid solution containing pepsin and taurodeoxycholic acid (TDCA) for 30 min, where Transepithelial Electrical Resistance (TER) was continuously measured. Control biopsies (no coated) were also exposed to acid solution. Then, the acid solution was replaced by the fluorescein solution, to assess transepithelial permeability for 90 min. In another experiment, the control and pretreated biopsies with AG at the concentration of 10% were exposed to acid solution pH 2.0 containing pepsin and TDCA for 120 min. The TER of the control biopsies and pretreated with AG-10% or sodium alginate was also evaluated after exposure to acid solution pH 1.0 containing pepsin and TDCA for 30 minutes, and then the acid solution was replaced by the fluorescein solution, for evaluation of the transepithelial permeability for 90 min. We performed an epifluorescence microscopy to localize on the oesopageal epithelium the biopolymer previously labeled with FITC. The pH 2.0 acid solution containing pepsin and TDCA promoted a gradual drop in TER in control biopsies of NERD patients and EE at the end of 30 min Pretreatment with AG-10% was able to prevent the drop in TER (P<0,05) compared to control biopsies. AG-1% and AG-3% did not protect the esophageal mucosa against the refluxate-like solution (P> 0.05). Acid exposure (pH 2.0 + TDCA + pepsin) on the luminal side of the esophageal mucosa of NERD and EE patients promoted a gradual increase in fluorescein permeability, with maximum effect over 90 min. Topical application of AG-10% significantly decreases (P <0.05) the fluorescein permeability in NERD patients, however, this reduction was not significant in EE patients (P> 0.05). The protective effect of AG-10% was sustained for up to 120 min both in biopsies of NERD and EE (P <0.05), when compared to control biopsies. AG-10% and sodium alginate were not able to protect biopsies against more acidic solution (pH 1.0) containing pepsin and TDCA when compared to control biopsies (P> 0.05). AG-10% and alginate were not able to decrease the fluorescein permeability when compared to the control group. Epifluorescent microscope images showed mucosal luminal adherence of FITC- labeled AG-10%. We concluded that AG-10% applied to mucosal biopsies of patients with NERD and EE has a prolonged topical protective effect against exposure to acid solution (pH 2.0 + TDCA + pepsin). However, this effect was not observed with the exposure of biopsies to more acidic solutions (pH 1.0 + TDCA + pepsin). These findings should be further explored in the development of new topical treatments for GERD patients.A doença do refluxo gastroesofágico (DRGE) é uma condição prevalente que afeta cerca de 13% da população mundial. Aproximadamente 60-70% dos pacientes não apresentam alterações macroscópicas no esófago, enquanto cerca de 20-30% apresentam erosões à endoscopia. Apesar da terapia com IBPs ser o tratamento de escolha, cerca de um terço dos pacientes não responde adequadamente ao tratamento, principalmente aqueles com doença não erosiva (NERD). Por isso formas alternativas de tratamento devem ser consideradas. Nosso objetivo foi avaliar a integridade da mucosa esofágica em pacientes com doença do refluxo gastroesofágico e investigar o efeito protetor tópico do polissacarídeo Anadenanthera colubrina (goma do angico, GA em sigla). Seis biópsias da mucosa do esôfago distal foram coletadas de 35 pacientes (24 sem erosão ou NERD e 11 com esofagite erosiva ou EE) e montadas em câmaras de Ussing. GA foi aplicada topicamente e em diferentes concentrações (1%, 3% e 10%, p/v), seguida de exposição com solução ácida pH 2,0 contendo pepsina e ácido taurodeoxicólico (TDCA) durante 30 min, onde a Resistência Elétrica Transepitelial (RET) foi continuamente medida. Biópsias-controle foram submetidas apenas a solução ácida. Em seguida, a solução ácida foi substituída pela solução de fluoresceína, para avaliação da permeabilidade transepitelial durante 90 min. Em outro experimento, as biopsias controle e pretratadas com GA na concentração de 10% foram desafiadas com solução ácida pH 2.0 contendo pepsina e TDCA durante 120 min. Também foi avaliada a RET das biopsias-controle e prétratadas com GA 10% ou com alginato de sódio após desafio com solução ácida pH 1.0 contendo pepsina e TDCA durante 30min, e em seguida a solução ácida foi substituída pela solução de fluoresceína, para avaliação da permeabilidade transepitelial durante 90 min. Realizamos um experimento utilizando técnicas de microscopia confocal para localizar no epitélio esofágico o biopolímero GA previamente marcado com FITC. A solução ácida pH 2.0 contendo pepsina e TDCA promoveu uma queda gradual da RET em biopsias-controle dos pacientes NERD e com EE ao final de 30 min. O pretratamento com GA a 10% foi capaz de prevenir a queda da RET (P<0,05) em comparação às biopsias-controle. GA 1% e GA 3% não foram capazes de proteger a mucosa esofágica contra a solução desafio (P > 0,05). A exposição ácida (pH 2.0 + TDCA + pepsina) no lado luminal da mucosa esofágica de pacientes NERD e EE promoveu um aumento gradual na permeabilidade à fluoresceína, com efeito máximo no tempo 90 min. A aplicação tópica da GA10% diminui significativamente (P<0,05) a passagem de fluoresceína nos pacientes NERD, porém essa redução não foi significativa nos pacientes EE (P > 0,05). O efeito protetor da GA 10% foi sustentado até 120 min tanto em biopsias dos pacientes NERD como em biopsias dos pacientes com EE (P < 0,05), quando comparadas às biopsias-controle. GA 10% e alginato de sódio não foram capazes de proteger as biopsias contra a solução ácida pH 1.0 contendo pepsina e TDCA em relação às biopsias-controle (P >0,05). GA10% e alginato não foram capazes de diminuir a passagem de fluoresceína (P >0,05) quando comparado ao grupo controle. As imagens do microscópio epifluorescente mostraram a aderência luminal à mucosa esofágica exercida pela GA10% marcado com FITC. Concluiu-se que GA10% aplicada a biópsias de mucosa de pacientes com NERD e EE tem um efeito tópico protetor in vitro prolongado contra a exposição à solução ácida (pH 2.0 + TDCA + pepsina). No entanto, esse efeito não foi observado com a exposição de biópsias a soluções muito ácidas (pH 1.0 + TDCA + pepsina). Esses achados devem ser mais explorados no desenvolvimento de novos tratamentos tópicos para pacientes com DRGE.Souza, Marcellus Henrique Loiola Ponte deBingana, Rudy Diavila2021-01-27T15:27:50Z2021-01-27T15:27:50Z2020-12-10info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfBINGANA, R. D. Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina. 2020. 80 f. Dissertação (Mestrado em Farmacologia) – Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2020.http://www.repositorio.ufc.br/handle/riufc/56218porreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccess2021-01-27T15:27:50Zoai:repositorio.ufc.br:riufc/56218Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:43:40.315395Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false |
dc.title.none.fl_str_mv |
Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina In vitro topical protection of esophageal mucosa from patients with gerd using “angico gum” a biopolymer from Anadenanthera colubrina |
title |
Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina |
spellingShingle |
Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina Bingana, Rudy Diavila Mucosa Esofágica Refluxo Gastroesofágico Gomas Vegetais |
title_short |
Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina |
title_full |
Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina |
title_fullStr |
Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina |
title_full_unstemmed |
Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina |
title_sort |
Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina |
author |
Bingana, Rudy Diavila |
author_facet |
Bingana, Rudy Diavila |
author_role |
author |
dc.contributor.none.fl_str_mv |
Souza, Marcellus Henrique Loiola Ponte de |
dc.contributor.author.fl_str_mv |
Bingana, Rudy Diavila |
dc.subject.por.fl_str_mv |
Mucosa Esofágica Refluxo Gastroesofágico Gomas Vegetais |
topic |
Mucosa Esofágica Refluxo Gastroesofágico Gomas Vegetais |
description |
Gastroesophageal reflux disease (GERD) is a prevalent condition that affects about 13% of the world population. Approximately 60-70% of patients do not present macroscopic changes in the esophagus, while about 20-30% have erosions on endoscopy. Although PPI therapy is the treatment of choice, about a third of patients do not respond adequately to treatment, especially those with Non-Erosive Reflux Disease (NERD). Therefore, alternative forms of treatment should be considered. We aimed to evaluate the integrity of the esophageal mucosa in patients with gastroesophageal reflux disease and to investigate the topical protective effect of polysaccharide Anadenanthera colubrina (Angico Gum, AG in acronym). Six biopsies of the distal esophageal mucosa were collected from 35 patients (24 without erosion or NERD and 11 with erosive esophagitis or EE) and mounted in Ussing chambers. AG was applied topically and in different concentrations (1%, 3% and 10%, w/v), followed by exposure with pH 2.0 acid solution containing pepsin and taurodeoxycholic acid (TDCA) for 30 min, where Transepithelial Electrical Resistance (TER) was continuously measured. Control biopsies (no coated) were also exposed to acid solution. Then, the acid solution was replaced by the fluorescein solution, to assess transepithelial permeability for 90 min. In another experiment, the control and pretreated biopsies with AG at the concentration of 10% were exposed to acid solution pH 2.0 containing pepsin and TDCA for 120 min. The TER of the control biopsies and pretreated with AG-10% or sodium alginate was also evaluated after exposure to acid solution pH 1.0 containing pepsin and TDCA for 30 minutes, and then the acid solution was replaced by the fluorescein solution, for evaluation of the transepithelial permeability for 90 min. We performed an epifluorescence microscopy to localize on the oesopageal epithelium the biopolymer previously labeled with FITC. The pH 2.0 acid solution containing pepsin and TDCA promoted a gradual drop in TER in control biopsies of NERD patients and EE at the end of 30 min Pretreatment with AG-10% was able to prevent the drop in TER (P<0,05) compared to control biopsies. AG-1% and AG-3% did not protect the esophageal mucosa against the refluxate-like solution (P> 0.05). Acid exposure (pH 2.0 + TDCA + pepsin) on the luminal side of the esophageal mucosa of NERD and EE patients promoted a gradual increase in fluorescein permeability, with maximum effect over 90 min. Topical application of AG-10% significantly decreases (P <0.05) the fluorescein permeability in NERD patients, however, this reduction was not significant in EE patients (P> 0.05). The protective effect of AG-10% was sustained for up to 120 min both in biopsies of NERD and EE (P <0.05), when compared to control biopsies. AG-10% and sodium alginate were not able to protect biopsies against more acidic solution (pH 1.0) containing pepsin and TDCA when compared to control biopsies (P> 0.05). AG-10% and alginate were not able to decrease the fluorescein permeability when compared to the control group. Epifluorescent microscope images showed mucosal luminal adherence of FITC- labeled AG-10%. We concluded that AG-10% applied to mucosal biopsies of patients with NERD and EE has a prolonged topical protective effect against exposure to acid solution (pH 2.0 + TDCA + pepsin). However, this effect was not observed with the exposure of biopsies to more acidic solutions (pH 1.0 + TDCA + pepsin). These findings should be further explored in the development of new topical treatments for GERD patients. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-12-10 2021-01-27T15:27:50Z 2021-01-27T15:27:50Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
BINGANA, R. D. Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina. 2020. 80 f. Dissertação (Mestrado em Farmacologia) – Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2020. http://www.repositorio.ufc.br/handle/riufc/56218 |
identifier_str_mv |
BINGANA, R. D. Proteção tópica in vitro da mucosa esofágica de pacientes com doença do refluxo gastroesofágico usando a “goma do angico”, um biopolímero de Anadenanthera colubrina. 2020. 80 f. Dissertação (Mestrado em Farmacologia) – Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2020. |
url |
http://www.repositorio.ufc.br/handle/riufc/56218 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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application/pdf |
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reponame:Repositório Institucional da Universidade Federal do Ceará (UFC) instname:Universidade Federal do Ceará (UFC) instacron:UFC |
instname_str |
Universidade Federal do Ceará (UFC) |
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UFC |
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UFC |
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Repositório Institucional da Universidade Federal do Ceará (UFC) |
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Repositório Institucional da Universidade Federal do Ceará (UFC) |
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Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC) |
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bu@ufc.br || repositorio@ufc.br |
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