P.11.4 ADHESION AND MUCOSAL EFFECT OF LACTOBACILLUS RHAMNOSUS GG IN INFLAMED AND NOT INFLAMED COLONIC MUCOSA EVALUATED BY AN EX-VIVO ORGAN CULTURE TECHNIQUE: A PROOF THE CONCEPT STUDY

ABSTRACT BACKGROUND The diarrheal syndrome is considered a serious public health problem all over the world and is considered a major cause of morbidity and mortality in developing countries. The high incidence of enteroaggregative Escherichia coli in diarrheal syndromes classified as an emerging pathogen of gastrointestinal infections. After decades of study, your pathogenesis remains uncertain and has been investigated mainly using in vitro models of adhesion in cellular lines. OBJECTIVE The present study investigated the interaction of enteroaggregative Escherichia coli strains isolated from childhood diarrhea with rabbit ileal and colonic mucosa ex vivo, using the in vitro organ culture model. METHODS The in vitro adhesion assays using cultured tissue were performed with the strains co-incubated with intestinal fragments of ileum and colon over a period of 6 hours. Each strain was tested with three intestinal fragments for each region. The fragments were analysed by scanning electron microscopy. RESULTS Through scanning electron microscopy we observed that all strains adhered to rabbit ileal and colonic mucosa, with the typical aggregative adherence pattern of “stacked bricks” on the epithelium. However, the highest degree of adherence was observed on colonic mucosa. Threadlike structures were found in greater numbers in the ileum compared to the colon. CONCLUSION These data showed that enteroaggregative Escherichia coli may have a high tropism for the human colon, which was ratified by the higher degree of adherence on the rabbit colonic mucosa. Finally, data indicated that in vitro organ culture of intestinal mucosa from rabbit may be used to elucidate the enteroaggregative Escherichia coli pathogenesis.

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Lactobacillus rhamnosus GG culture supernatant ameliorates acute alcohol-induced intestinal permeability and liver injury

AJP Gastrointestinal and Liver Physiology ◽

10.1152/ajpgi.00024.2012 ◽

2012 ◽

Vol 303 (1) ◽

pp. G32-G41 ◽

Cited By ~ 114

Author(s):

Yuhua Wang ◽

Yanlong Liu ◽

Anju Sidhu ◽

Zhenhua Ma ◽

Craig McClain ◽

...

Keyword(s):

Liver Injury ◽

Intestinal Permeability ◽

Culture Supernatant ◽

Ex Vivo ◽

Liver Steatosis ◽

Lactobacillus Rhamnosus ◽

Intestinal Barrier ◽

Chow Diet ◽

Mrna Levels ◽

Lactobacillus Rhamnosus Gg

Endotoxemia is a contributing cofactor to alcoholic liver disease (ALD), and alcohol-induced increased intestinal permeability is one of the mechanisms of endotoxin absorption. Probiotic bacteria have been shown to promote intestinal epithelial integrity and protect barrier function in inflammatory bowel disease (IBD) and in ALD. Although it is highly possible that some common molecules secreted by probiotics contribute to this action in IBD, the effect of probiotic culture supernatant has not yet been studied in ALD. We examined the effects of Lactobacillus rhamnosus GG culture supernatant (LGG-s) on the acute alcohol-induced intestinal integrity and liver injury in a mouse model. Mice on standard chow diet were supplemented with supernatant from LGG culture (109 colony-forming unit/mouse) for 5 days, and one dose of alcohol at 6 g/kg body wt was administered via gavage. Intestinal permeability was measured by FITC-FD-4 ex vivo. Alcohol-induced liver injury was examined by measuring the activity of alanine aminotransferase (ALT) in plasma, and liver steatosis was evaluated by triglyceride content and Oil Red O staining of the liver sections. LGG-s pretreatment restored alcohol-induced reduction in ileum mRNA levels of claudin-1, intestine trefoil factor (ITF), P-glycoprotein (P-gp), and cathelin-related antimicrobial peptide (CRAMP), which play important roles on intestinal barrier integrity. As a result, LGG-s pretreatment significantly inhibited the alcohol-induced intestinal permeability, endotoxemia and subsequently liver injury. Interestingly, LGG-s pretreatment increased ileum mRNA expression of hypoxia-inducible factor (HIF)-2α, an important transcription factor of ITF, P-gp, and CRAMP. These results suggest that LGG-s ameliorates the acute alcohol-induced liver injury by promoting HIF signaling, leading to the suppression of alcohol-induced increased intestinal permeability and endotoxemia. The use of bacteria-free LGG culture supernatant provides a novel strategy for prevention of acute alcohol-induced liver injury.

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P.12.8 EVALUATION OF PROBIOTIC BACTERIA ADHESION TO NORMAL AND DYSPLASTIC COLONIC MUCOSA BY AN EX-VIVO ORGAN CULTURE EXPERIMENTAL MODEL

Digestive and Liver Disease ◽

10.1016/s1590-8658(13)60497-1 ◽

2013 ◽

Vol 45 ◽

pp. S175

Author(s):

C. Pagnini ◽

G. Rizzatti ◽

V. Corleto ◽

S. Angeletti ◽

E. Di Giulio ◽

...

Keyword(s):

Organ Culture ◽

Experimental Model ◽

Colonic Mucosa ◽

Ex Vivo ◽

Probiotic Bacteria ◽

Bacteria Adhesion

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Physicochemical Investigation of Biosynthesis of a Protein Coating on Glass That Promotes Mammalian Cell Growth Using Lactobacillus rhamnosus GG Bacteria

Coatings ◽

10.3390/coatings11111410 ◽

2021 ◽

Vol 11 (11) ◽

pp. 1410

Author(s):

Kamil Kaminski ◽

Karolina Syrek ◽

Joanna Grudzień ◽

Magdalena Obloza ◽

Monika Adamczyk ◽

...

Keyword(s):

Cell Growth ◽

Mammalian Cell ◽

Mammalian Cells ◽

Ex Vivo ◽

Lactobacillus Rhamnosus ◽

Vero Cells ◽

Lactobacillus Rhamnosus Gg ◽

Composition And Structure ◽

Mammalian Cell Growth ◽

Protein Components

Glass surfaces, although the first to be used for culturing ex vivo adherent cells, are not the perfect substrates for this purpose. Today, plastics dominate these applications, but in light of the global trend to reduce the use of synthetic polymers, it is reasonable to consider a return to glass vessels with coatings for these purposes. The ideal surface for cell growth is one that simulates the composition and structure of the mainly protein-based intercellular matrix. The work presented here shows a new idea of preparing porous protein coatings on glass using biosynthesis. The process utilizes the colonization of the gold nanoparticle-coated glass surface with Lactobacillus rhamnosus GG bacteria, followed by permeabilization (using ethanol) of their membrane and partial thermal degradation (at 160 °C in vacuum) of the surface-bound protein components of these microorganisms. It results in a development of coating on the glass that promotes mammalian cell growth, which has been preliminary confirmed using Vero cells. Subsequent steps in the formation of coating components were documented by reflectance ultraviolet and visible spectra and infrared spectroscopy. The presence of microorganisms and mammalian cells was confirmed using scanning electron and optical microscopy and crystalline violet staining.

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