Induction of human-chorioamniotic-membrane remodeling in-vitro by the alpha hemolysin of Escherichia coli
Introduction: Almost 80% of urinary tract infections during pregnancy are caused by uropathogenic strains of Escherichia coli. Alpha-hemolysin, toxin secreted by them, has a fundamental role in this pathology development. Considering that urinary tract infections are related with premature rupture...
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| Formato: | Articulo Preprint |
| Lenguaje: | Inglés |
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2024
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| Acceso en línea: | http://sedici.unlp.edu.ar/handle/10915/162844 |
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I19-R120-10915-1628442024-02-19T20:09:07Z http://sedici.unlp.edu.ar/handle/10915/162844 Induction of human-chorioamniotic-membrane remodeling in-vitro by the alpha hemolysin of Escherichia coli Pucci Molineris, Melisa Eliana Schibert, Florencia Lima, María Silvia Accialini, Paula Cané, Lucía Pelinski, Pablo Farina, Mariana Herlax, Vanesa Silvana 2024-02-13 2024-02-19T14:57:41Z en Ciencias Médicas hemolysin fetal membranes. PROM urinary-tract infections uropathogenic strains virulence factor Introduction: Almost 80% of urinary tract infections during pregnancy are caused by uropathogenic strains of Escherichia coli. Alpha-hemolysin, toxin secreted by them, has a fundamental role in this pathology development. Considering that urinary tract infections are related with premature rupture of fetal membranes, we proposed to evaluate the effects that alpha-hemolysin induces on human-fetal-membranes. Methods: Thirteen fetal membranes obtained from elective cesarean sections (>37 weeks) were mounted in a transwell-device generating two independent chambers. To mimic an ascendant-urinary-tract infection, membranes were incubated with different concentrations of pure alpha-hemolysin from the choriodecidual side during 24h. Extensive histological analyses were performed and transepithelial electrical-resistance were determined. Cell viability, metalloproteinase activity and cyclooxygenase-2- gene expression was estimated by lactate-dehydrogenase–release assay, zymography and RT-qPCR, respectively. Finally, four fetal membranes were treated with hemolysin preincubated with polyclonal anti-hemolysin antibodies. Cell viability and metalloproteinase activity were monitored. Results: After 24 h of treatment, fetal membranes evidenced a structural damage and a decrease in membrane resistance that progressed as the concentration of alpha hemolysin increased. While the amniotic-epithelial-layer remained practically unaffected, the chorion cells manifested an increase in vacuolization and necrosis. In addition, the extracellular matrix exhibited collagen-fiber disorganization, a marked decrease in fiber content, and became thicker in presence of the toxin. Cyclooxigenase-2 expression and metalloproteinase activity were also higher in the treated groups than in untreated ones. Finally, a preincubation of hemolysin with specific antibodies prevented the cytotoxicity on the chorion cells and the increase in metalloproteinase activity. Discussion: Hemolysin induces structural and molecular changes associated with the remodeling of human-fetal-membranes in-vitro. Instituto de Investigaciones Bioquímicas de La Plata Facultad de Ciencias Médicas Articulo Preprint http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf |
| institution |
Universidad Nacional de La Plata |
| institution_str |
I-19 |
| repository_str |
R-120 |
| collection |
SEDICI (UNLP) |
| language |
Inglés |
| topic |
Ciencias Médicas hemolysin fetal membranes. PROM urinary-tract infections uropathogenic strains virulence factor |
| spellingShingle |
Ciencias Médicas hemolysin fetal membranes. PROM urinary-tract infections uropathogenic strains virulence factor Pucci Molineris, Melisa Eliana Schibert, Florencia Lima, María Silvia Accialini, Paula Cané, Lucía Pelinski, Pablo Farina, Mariana Herlax, Vanesa Silvana Induction of human-chorioamniotic-membrane remodeling in-vitro by the alpha hemolysin of Escherichia coli |
| topic_facet |
Ciencias Médicas hemolysin fetal membranes. PROM urinary-tract infections uropathogenic strains virulence factor |
| description |
Introduction: Almost 80% of urinary tract infections during pregnancy are caused by uropathogenic strains of Escherichia coli. Alpha-hemolysin, toxin secreted by them, has a fundamental role in this pathology development. Considering that urinary tract infections are related with premature rupture of fetal membranes, we proposed to evaluate the effects that alpha-hemolysin induces on human-fetal-membranes.
Methods: Thirteen fetal membranes obtained from elective cesarean sections (>37 weeks) were mounted in a transwell-device generating two independent chambers. To mimic an ascendant-urinary-tract infection, membranes were incubated with different concentrations of pure alpha-hemolysin from the choriodecidual side during 24h. Extensive histological analyses were performed and transepithelial electrical-resistance were determined. Cell viability, metalloproteinase activity and cyclooxygenase-2- gene expression was estimated by lactate-dehydrogenase–release assay, zymography and RT-qPCR, respectively. Finally, four fetal membranes were treated with hemolysin preincubated with polyclonal anti-hemolysin antibodies. Cell viability and metalloproteinase activity were monitored.
Results: After 24 h of treatment, fetal membranes evidenced a structural damage and a decrease in membrane resistance that progressed as the concentration of alpha hemolysin increased. While the amniotic-epithelial-layer remained practically unaffected, the chorion cells manifested an increase in vacuolization and necrosis. In addition, the extracellular matrix exhibited collagen-fiber disorganization, a marked decrease in fiber content, and became thicker in presence of the toxin. Cyclooxigenase-2 expression and metalloproteinase activity were also higher in the treated groups than in untreated ones. Finally, a preincubation of hemolysin with specific antibodies prevented the cytotoxicity on the chorion cells and the increase in metalloproteinase activity.
Discussion: Hemolysin induces structural and molecular changes associated with the remodeling of human-fetal-membranes in-vitro. |
| format |
Articulo Preprint |
| author |
Pucci Molineris, Melisa Eliana Schibert, Florencia Lima, María Silvia Accialini, Paula Cané, Lucía Pelinski, Pablo Farina, Mariana Herlax, Vanesa Silvana |
| author_facet |
Pucci Molineris, Melisa Eliana Schibert, Florencia Lima, María Silvia Accialini, Paula Cané, Lucía Pelinski, Pablo Farina, Mariana Herlax, Vanesa Silvana |
| author_sort |
Pucci Molineris, Melisa Eliana |
| title |
Induction of human-chorioamniotic-membrane remodeling in-vitro by the alpha hemolysin of Escherichia coli |
| title_short |
Induction of human-chorioamniotic-membrane remodeling in-vitro by the alpha hemolysin of Escherichia coli |
| title_full |
Induction of human-chorioamniotic-membrane remodeling in-vitro by the alpha hemolysin of Escherichia coli |
| title_fullStr |
Induction of human-chorioamniotic-membrane remodeling in-vitro by the alpha hemolysin of Escherichia coli |
| title_full_unstemmed |
Induction of human-chorioamniotic-membrane remodeling in-vitro by the alpha hemolysin of Escherichia coli |
| title_sort |
induction of human-chorioamniotic-membrane remodeling in-vitro by the alpha hemolysin of escherichia coli |
| publishDate |
2024 |
| url |
http://sedici.unlp.edu.ar/handle/10915/162844 |
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