The Co-Culture of Staphylococcal Biofilm and Fibroblast Cell Line: The Correlation of Biological Phenomena with Metabolic NMR1 Footprint.

Joanna Czajkowska,Andrzej Pawlak,Monika Toporkiewicz,Adam Junka,Marcin Śmiglak,Jakub Hoppe,Karol Fijałkowski,Paweł Migdał,Monika Oleksy-Wawrzyniak,Agata Markowska-Szczupak

doi:10.3390/ijms22115826

Abstract

Staphylococcus aureus is one of the most prevalent pathogens associated with several types of biofilm-based infections, including infections of chronic wounds. Mature staphylococcal biofilm is extremely hard to eradicate from a wound and displays a high tendency to induce recurring infections. Therefore, in the present study, we aimed to investigate in vitro the interaction between S. aureus biofilm and fibroblast cells searching for metabolites that could be considered as potential biomarkers of critical colonization and infection. Utilizing advanced microscopy and microbiological methods to examine biofilm formation and the staphylococcal infection process, we were able to distinguish 4 phases of biofilm development. The analysis of staphylococcal biofilm influence on the viability of fibroblasts allowed us to pinpoint the moment of critical colonization—12 h post contamination. Based on the obtained model we performed a metabolomics analysis by 1H NMR spectroscopy to provide new insights into the pathophysiology of infection. We identified a set of metabolites related to the switch to anaerobic metabolism that was characteristic for staphylococcal biofilm co-cultured with fibroblast cells. The data presented in this study may be thus considered a noteworthy but preliminary step in the direction of developing a new, NMR-based tool for rapid diagnosing of infection in a chronic wound.

Highlights

The treatment of infected chronic wounds remains a global challenge for health care systems and consumes about 1–3% of European Union public health budgets [1].Regardless of the type, if managed inadequately, chronic wounds cause a dramatic deterioration of patients’ life quality [2].the complications caused by microbial biofilm settled within chronic wounds pose a risk of amputation, development of a systemic infection and, as a consequence, may lead to a deterioration of a patient’s health or even their death [3].Devoid of the skin’s protective barrier and repeatedly flooded with nutrient-rich exudate, chronic wounds are attractive niches for microbial colonization and formation of biofilm
In the first step of the investigation, we have conducted a series of experiments to analyze biofilm process of aureus
Chronic wounds, which are evident examples of such diseases, are an enormous challenge for medical care because they frequently occur secondary to such other diseases as diabetes, cardiovascular diseases, obesity or cancers [15,16,17]

Summary

Introduction

The treatment of infected chronic wounds remains a global challenge for health care systems and consumes about 1–3% of European Union public health budgets [1].Regardless of the type (diabetic foot, burns, venous leg ulcers, pressure ulcers), if managed inadequately, chronic wounds cause a dramatic deterioration of patients’ life quality [2].the complications caused by microbial biofilm settled within chronic wounds pose a risk of amputation, development of a systemic infection and, as a consequence, may lead to a deterioration of a patient’s health or even their death [3].Devoid of the skin’s protective barrier and repeatedly flooded with nutrient-rich exudate, chronic wounds are attractive niches for microbial colonization and formation of biofilm. The treatment of infected chronic wounds remains a global challenge for health care systems and consumes about 1–3% of European Union public health budgets [1]. Regardless of the type (diabetic foot, burns, venous leg ulcers, pressure ulcers), if managed inadequately, chronic wounds cause a dramatic deterioration of patients’ life quality [2]. The complications caused by microbial biofilm settled within chronic wounds pose a risk of amputation, development of a systemic infection and, as a consequence, may lead to a deterioration of a patient’s health or even their death [3]. Devoid of the skin’s protective barrier and repeatedly flooded with nutrient-rich exudate, chronic wounds are attractive niches for microbial colonization and formation of biofilm. The results of recent studies indicate that biofilm is the dominant form of microbial existence in wounds [5]

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Conclusion