Evaluation of In Vitro Biofilm Formation of Helicobacter pylori in Different Culture Media

Year 2024, , 98 - 104, 01.11.2024

Sibel Gülseroğlu , Hale Akpınar Özlem Yılmaz , Müjde Soytürk

https://doi.org/10.38175/phnx.1455085

Abstract

Objective: Biofilms are surface-attached cell communities that play a role in the survival of bacteria. Helicobacter pylori, a gram-negative pathogen that colonizes the human gastric mucosa, forms biofilms, causing treatment failure and the risk of developing peptic ulcers, gastritis and gastric cancer in infected individuals. The aim of the study is to evaluate the biofilm formation abilities of H. pylori ATCC 26695 and three clinical strains in different culture media.
Material and Method: Biofilm formation characteristics of H. pylori strains using different culture media were evaluated, and the crystal violet (CV) staining method (measured at OD 595 ) was used. Various media were used for incubating H. pylori strains: Brucella broth (BB), TSB with 10% FBS, BHI with 10% FBS, BB with 10% FBS, BB with 10% FBS + 0.25% glucose, and BB with 10% FBS + 1% glucose (incubated for 3 days). Additionally, BB with 1% FBS, BB with 10% FBS, and BB with 5% inactivated human serum were incubated for 2 and 4 days at 37°C under microaerophilic conditions.
Results: It was observed that 5% inactived human serum was more effective in biofilm formation of H. pylori ATCC 26695 and three clinical strains. However, there was no biofilm production in the strains cultured with Brucella broth alone and that the strains cultured with TSB + 10% FBS could not form a strong biofilm compared to other media.
Conclusion: Different culture media used for H. pylori ATCC 26695 and three clinical strains affect biofilm formation. It is thought that in vitro experiments to prevent biofilm formation may provide a solution to the prevention of H. pylori infection.

Keywords

biofilm;, Helicobacter pylori, ATCC 26695;, infection;, crystal violet

Ethical Statement

Dokuz Eylül University Ethics Committee

Supporting Institution

Dokuz Eylül University Scientific Research Projects Commission Unit

Project Number

Decision No: 2022/04-16, Date 20.07.2022

Helicobacter Pylori Farklı Kültür Ortamlarında İn Vitro Biyofilm Oluşumunun Değerlendirilmesi

Abstract

Amaç: Biyofilmler, bakterilerin hayatta kalmasında rol oynayan yüzeye bağlı hücre topluluklarıdır. İnsan mide mukozasını kolonize eden gram negatif bir patojen olan Helicobacter pylori, biyofilm oluşturarak tedavi başarısızlığına ve enfekte bireylerde peptik ülser, gastrit ve mide kanseri gelişme riskine neden olur. H. pylori ATCC 26695 ve üç klinik suşun farklı kültür ortamlarında biyofilm oluşturma yeteneklerinin değerlendirilmesi amaçlanmıştır.
Gereç ve Yöntem: H. pylori suşlarının biyofilm oluşturma özellikleri farklı kültür ortamları kullanılarak biyofilm oluşturmaları Kristal viyole (CV) yöntemi ile spektrofotometrede (OD595) değerlendirildi. Kullanılan ortamlar; Brucella broth (BB) besiyeri, TSB + %10 FBS, BHI + %10 FBS, BB + %10 FBS, BB + %10 FBS + %0.25 glukoz ve BB + %10 FBS + %1 glukoz-(3 gün inkübasyon) ve BB + %1 FBS, BB + %10 FBS, BB + %5 inaktive insan serumu 2 ve 4 gün 37°C de mikroaerofilik koşullarda inkübasyon gerçekleştirildi.
Bulgular: H. pylori ATCC 26695 ve üç klinik suşun biyofilm oluşumunda %5 inaktive edilmiş insan serumunun daha etkili olduğu görüldü. Sadece Brucella besiyeri ile biyofilm üretiminin olmadığı TSB + %10 FBS ile suşların diğer ortamlara göre güçlü bir biyofilm oluşturmadığı gözlendi.
Sonuç: H. pylori ATCC 26695 ve üç klinik suş için kullanılan farklı kültür ortamları biyofilm oluşumunu etkilemektedir. Biyofilm gelişiminin önlenmesine yönelik in vitro deneylerin yapılmasının H. pylori enfeksiyonlarının önlenmesinde çözümler sağlayabileceği düşünülmektedir.

Keywords

ATCC 26695, Biyofilm, Kristal Viyole, Helicobacter pylori, Enfeksiyon

Project Number

Decision No: 2022/04-16, Date 20.07.2022

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