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

Czajkowska, Joanna; Junka, Adam; Hoppe, Jakub; Toporkiewicz, Monika; Pawlak, A.L.; Migdał, Paweł; Oleksy-Wawrzyniak, Monika; Fijałkowski, Karol; Śmiglak, Marcin; Markowska‐Szczupak, Agata

doi:10.3390/ijms22115826

Cited by 8 publications

(6 citation statements)

References 47 publications

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“…They highlighted how these complementary analyses provided information on intracellular and extracellular targets, such as protein, DNA and cell wall. Czajkowska et al [ 36 ] employed 1 H NMR extracellular footprinting analysis to investigate in vitro interactions between S. aureus biofilm and fibroblasts. However, all these studies were performed as single measurements.…”

Section: Discussionmentioning

confidence: 99%

Determination of Intra- and Extracellular Metabolic Adaptations of 3D Cell Cultures upon Challenges in Real-Time by NMR

Urzì

Hertig

Meyer

et al. 2022

IJMS

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NMR flow devices provide longitudinal real-time quantitative metabolome characterisation of living cells. However, discrimination of intra- and extracellular contributions to the spectra represents a major challenge in metabolomic NMR studies. The present NMR study demonstrates the possibility to quantitatively measure both metabolic intracellular fingerprints and extracellular footprints on human control fibroblasts by using a commercially available flow tube system with a standard 5 mm NMR probe. We performed a comprehensive 3D cell culture system characterisation. Diffusion NMR was employed for intra- and extracellular metabolites separation. In addition, complementary extracellular footprints were determined. The implemented perfused NMR bioreactor system allowed the determination of 35 metabolites and intra- and extracellular separation of 19 metabolites based on diffusion rate differences. We show the reliability and sensitivity of NMR diffusion measurements to detect metabolite concentration changes in both intra- and extracellular compartments during perfusion with different selective culture media, and upon complex I inhibition with rotenone. We also demonstrate the sensitivity of extracellular footprints to determine metabolic variations at different flow rates. The current method is of potential use for the metabolomic characterisation of defect fibroblasts and for improving physiological comprehension.

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Section: Discussionmentioning

confidence: 99%

Determination of Intra- and Extracellular Metabolic Adaptations of 3D Cell Cultures upon Challenges in Real-Time by NMR

Urzì

Hertig

Meyer

et al. 2022

IJMS

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“…The viability and proliferation of the microbial cells were evaluated using quantitative culturing on the appropriate solid agar plates (Müller-Hinton agar for bacteria and Sabouraud agar for yeast (Biomaxima, Lublin, Poland)) and tetrazolium test every 24 hours for 3 days. The 24 h co-culture of microbes and fibroblasts was chosen as a model for antiseptic application, similarly as we have shown it earlier ( Czajkowska et al., 2021 ). Such prepared biofilm grown on the fibroblast-containing BC carrier was exposed to 2 mL of P, G or M antiseptics for 1 hour of contact time.…”

Section: Methodsmentioning

confidence: 99%

Comparison of antibiofilm activity of low-concentrated hypochlorites vs polyhexanide-containing antiseptic

Paleczny

Junka²,

Krzyżek³

et al. 2023

Front. Cell. Infect. Microbiol.

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Chronic wound infection is highly associated with morbidity and endangers the patient's life. Therefore, wound care products must have a potent antimicrobial and biofilm-eradicating effect. In this work, the antimicrobial/antibiofilm activity of two low-concentrated chlorine-based and releasing solutions was investigated on a total of 78 strains of methicillin-resistant Staphylococcus aureus, Pseudomonas aeruginosa, and Candida albicans, using the cohesive spectrum of in vitro settings, including microtiter plate models, biofilm-oriented antiseptic test, cellulose-based biofilm model, biofilm bioreactors and Bioflux model. The antiseptic containing polyhexamethylene biguanide was used in the character of usability control of performed tests. The results obtained by static biofilm models indicate that low-concentrated chlorine-based and releasing solutions display none to moderate antibiofilm activity, while data obtained by means of the Bioflux model, providing flow conditions, indicate the moderate antibiofilm activity of substances compared with the polyhexanide antiseptic. Considering in vitro data presented in this manuscript, the earlier reported favorable clinical results of low-concentrated hypochlorites should be considered rather an effect of their rinsing activity combined with low cytotoxicity but not the antimicrobial effect per se. For the treatment of heavily biofilm-infected wounds, polyhexanide should be considered the agent of choice because of its higher efficacy against pathogenic biofilms.

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“…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 [4] . The results of recent studies indicate that biofilm is the dominant form of microbial existence in wounds [5,6] . Biofilm is frequently defined based on in vitro observations.…”

Section: Introductionmentioning

confidence: 99%

Visual analysis of chronic wounds and its correlation with biofilm

Choudhary¹,

Pandey²,

Murty³

et al. 2022

Int. J. Surg. Sci.

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To evaluate the biofilm using visual indicators that were clinically observable to see if any significant method of clinically detecting or predicting the presence of a biofilm can be developed. Material and methods:The prospective study was conducted from late 2019 to 2021 in the Department of Surgery at Chatrapati Shivaji Subharti Hospital, a tertiary care super speciality institute of Swami Vivekanand Subharti University among patients with chronic wounds admitted in the department through surgery outpatient department/Emergency/transferred from other departments formed the study group. Samples were subjected for aerobic bacterial culture on blood agar, chocolate agar and MacConkey agar plates. The isolated bacterial pathogen was identified using standard bacteriological procedures which included colony morphology, gram stain and battery biochemical tests both for Gram positive and negative bacteria as per Gram stain. Efficacy of visual score was calculated using diagnostic tests (Sensitivity, Specificity, Diagnostic Accuracy) considering biofilm assessment by culture method as gold standard. Results: Visual score viz. 0 (None), ≤5 (Not Sure), 6-8 (Probable) and ≥9 (Predicted) was reported in 0%, 8%, 31% and 61% of the subjects respectively. Sensitivity, specificity, positive predictive value, negative predictive value and accuracy rate of visual score considering culture method as gold standard was 94.74%, 83.72%, 99.10%, 45.57% and 94.19% respectively. Conclusion:We concluded from our findings that clinical algorithm (visual score) may serve as a nearly signal to alert clinicians that the wound has diverted off its normal healing path. Detection of early (young) biofilms enable intervention before recalcitrance and/or infection become a problem.

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

Cited by 8 publications

References 47 publications

Determination of Intra- and Extracellular Metabolic Adaptations of 3D Cell Cultures upon Challenges in Real-Time by NMR

Determination of Intra- and Extracellular Metabolic Adaptations of 3D Cell Cultures upon Challenges in Real-Time by NMR

Comparison of antibiofilm activity of low-concentrated hypochlorites vs polyhexanide-containing antiseptic

Visual analysis of chronic wounds and its correlation with biofilm

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