title: Effect of gastric environment on Helicobacter pylori adhesion to a mucoadhesive polymer.
abstract: Helicobacter pylori infection has been associated with several gastric diseases. This bacterium colonizes the gastric mucosa of half of the world's population, and available treatments are unsuccessful in practically one in every five patients. Mucoadhesive polymers, such as chitosan, are being investigated as gastric drug delivery systems. However, since chitosan is also known for its antimicrobial properties, this work aims to evaluate H. pylori interactions with chitosan under simulated gastric environments, namely using various pHs (2.6, 4 and 6), pepsin and urea. To enable the visualization of adherent bacteria, ultrathin chitosan films were produced by spin-coating on gold/glass surfaces, cross-linked with genipin and characterized by Fourier transform infrared reflection absorption spectroscopy, ellipsometry and electrokinetic analysis. Films with homogeneous thickness of 11.7±0.6 nm were produced, and were stable and protonated at all the pHs used. Furthermore, they adsorbed pepsin in all these pHs, in contrast to urea, of which a small adsorption was only observed at pH 6. H. pylori binding to chitosan was higher at pH2.6 although most of adherent bacteria were dead. The presence of pepsin decreased bacterial adhesion, but increased its viability while in a more stressed morphology (coccoid form). The presence of urea did not affect the amount, morphology or viability of chitosan-adherent bacteria. In suspension, the decrease in pH changed H. pylori zeta potential from negative to positive. Moreover, bacteria were only culturable when incubated in pH 6 with and without urea (without pepsin). This work demonstrates that chitosan has the capacity to bind and kill H. pylori in a range of pHs independently of urea. This opens new perspectives for the application of chitosan-based materials to the elimination of H. pylori gastric colonization, though pepsin might appear to be an obstacle.
authors.affiliation: INEB-Instituto de Engenharia Biomédica, Universidade do Porto, Rua do Campo Alegre 823, 4150-180 Porto, Portugal.
authors.forename: Frederico
authors.initials: F
authors.lastname: Nogueira
authors.forename: Inês C
authors.initials: IC
authors.lastname: Gonçalves
authors.forename: M Cristina L
authors.initials: MC
authors.lastname: Martins
mesh_terms: D000268:Adhesiveness / Q000187:drug effects; D001422:Bacterial Adhesion / Q000187:drug effects*; D002021:Buffers; D048271:Chitosan / Q000494:pharmacology*; D016480:Helicobacter pylori / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D006863:Hydrogen-Ion Concentration; D050296:Microbial Viability / Q000187:drug effects; D009093:Mucus / Q000737:chemistry*; D010434:Pepsin A / Q000494:pharmacology; D017550:Spectroscopy, Fourier Transform Infrared; D013270:Stomach / Q000382:microbiology*; D014508:Urea / Q000494:pharmacology
chemical_list: D002021:Buffers; D014508:Urea; D048271:Chitosan; D010434:Pepsin A