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Negative Pressure Level and Effects on Bacterial Growth Kinetics in an in vitro Wound Model

Adam Bobkiewicz

Adam Bobkiewicz

Department of General, Endocrinological Surgery and Gastroenterological Oncology Poznan, University of Medical SciencesPoznan, Poland
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Bobkiewicz, Adam
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Wojciech Francuzik

Wojciech Francuzik

Department of Dermatology, Venerology and Allergology. Charité – Universitätsmedizin BerlinBerlin, Germany
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Francuzik, Wojciech
, 
Amy Martinkosky

Amy Martinkosky

Department of General, Endocrinological Surgery and Gastroenterological Oncology Poznan, University of Medical SciencesPoznan, Poland
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Martinkosky, Amy
, 
Maciej Borejsza-Wysocki

Maciej Borejsza-Wysocki

Department of General, Endocrinological Surgery and Gastroenterological Oncology Poznan, University of Medical SciencesPoznan, Poland
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Borejsza-Wysocki, Maciej
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Witold Ledwosinski

Witold Ledwosinski

Department of General, Endocrinological Surgery and Gastroenterological Oncology Poznan, University of Medical SciencesPoznan, Poland
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Ledwosinski, Witold
, 
Krzysztof Szmyt

Krzysztof Szmyt

Department of General, Endocrinological Surgery and Gastroenterological Oncology Poznan, University of Medical SciencesPoznan, Poland
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Szmyt, Krzysztof
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Tomasz Banasiewicz

Tomasz Banasiewicz

Department of General, Endocrinological Surgery and Gastroenterological Oncology Poznan, University of Medical SciencesPoznan, Poland
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Banasiewicz, Tomasz
 e 
Lukasz Krokowicz

Lukasz Krokowicz

Department of General, Endocrinological Surgery and Gastroenterological Oncology Poznan, University of Medical SciencesPoznan, Poland
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Krokowicz, Lukasz
  
20 giu 2024
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Categoria dell'articolo: Original Paper
Pubblicato online: 20 giu 2024
Pagine: 199 - 206
Ricevuto: 18 feb 2024
Accettato: 20 apr 2024
DOI: https://doi.org/10.33073/pjm-2024-018
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© 2024 Adam Bobkiewicz et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
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Fig. 1.

Detailed representation of a custom-engineered acrylic negative pressure chamber for skin wound model examination. The figure depicts an overhead view of an individually numbered negative pressure chamber designated for studying the impacts of vacuum conditions on bacterial colonization in a skin wound model. The principal compartment, ‘E’, serves as the receptacle for the skin wound model. The application of negative pressure is facilitated through the portal marked ‘C’, hermetically sealed with a silicone gasket ‘B’. The electronic pressure sensor ‘D’ relays the internal pressure reading of −0.26 atm, signifying an operational negative pressure state. This sensor is linked via cable ‘H’ to an external device for continuous pressure monitoring. Port ‘F’ is utilized for instillation, permitting the administration or removal of substances without nullifying the vacuum environment. Access knobs ‘A’ allow for the manipulation of the internal contents. The device’s identification is presented on the label ‘G’, indicating the unique chamber number ranging from 01 to 24, essential for distinguishing among multiple units in use during parallel experiments. This system is fundamental for the precise study of bacterial dynamics under negative pressure, a critical factor in optimizing NPWT protocols for enhanced wound healing outcomes.
Detailed representation of a custom-engineered acrylic negative pressure chamber for skin wound model examination. The figure depicts an overhead view of an individually numbered negative pressure chamber designated for studying the impacts of vacuum conditions on bacterial colonization in a skin wound model. The principal compartment, ‘E’, serves as the receptacle for the skin wound model. The application of negative pressure is facilitated through the portal marked ‘C’, hermetically sealed with a silicone gasket ‘B’. The electronic pressure sensor ‘D’ relays the internal pressure reading of −0.26 atm, signifying an operational negative pressure state. This sensor is linked via cable ‘H’ to an external device for continuous pressure monitoring. Port ‘F’ is utilized for instillation, permitting the administration or removal of substances without nullifying the vacuum environment. Access knobs ‘A’ allow for the manipulation of the internal contents. The device’s identification is presented on the label ‘G’, indicating the unique chamber number ranging from 01 to 24, essential for distinguishing among multiple units in use during parallel experiments. This system is fundamental for the precise study of bacterial dynamics under negative pressure, a critical factor in optimizing NPWT protocols for enhanced wound healing outcomes.

Fig. 2.

Influence of negative pressure on Staphylococcus aureus and Staphylococcus epidermidis growth at different negative pressure levels. This composite figure presents a series of box-and-whisker plots alongside density plots demonstrating the effects of varying levels of negative pressure on the growth characteristics of S. aureus and S. epidermidis. Each row represents a different combination of bacterial species and culture duration, with panel A corresponding to S. aureus at 120 hours culture, panel B to S. epidermidis at 120 hours culture. The first column of box plots in each panel depicts the colony-forming units (CFU), the second shows the mean area per colony, and the third illustrates the total growth area across a range of negative pressures from −50 to −250 mmHg. The accompanying density plots visualize the distribution of colony areas, with vertical dashed lines marking the cut-off values for debris.
Influence of negative pressure on Staphylococcus aureus and Staphylococcus epidermidis growth at different negative pressure levels. This composite figure presents a series of box-and-whisker plots alongside density plots demonstrating the effects of varying levels of negative pressure on the growth characteristics of S. aureus and S. epidermidis. Each row represents a different combination of bacterial species and culture duration, with panel A corresponding to S. aureus at 120 hours culture, panel B to S. epidermidis at 120 hours culture. The first column of box plots in each panel depicts the colony-forming units (CFU), the second shows the mean area per colony, and the third illustrates the total growth area across a range of negative pressures from −50 to −250 mmHg. The accompanying density plots visualize the distribution of colony areas, with vertical dashed lines marking the cut-off values for debris.

Fig. 3.

Influence of negative pressure on Staphylococcus aureus growth at different negative pressure intervals. This composite figure presents a series of box-and-whisker plots alongside density plots demonstrating the effects of varying levels of negative pressure on the growth characteristics of Staphylococcus aureus using different modes of intermittent negative pressure intervals. The first column of box plots in each panel depicts the colony-forming units (CFU), and the second illustrates the total growth area, across an intermittent negative pressure intervals of 1–3 h and the second illustrates the total growth area across intermittent negative pressure intervals of 1–3 h. The pressure used was 250 mmHg. The pressure used was 250 mmHg. The accompanying density plots visualize the distribution of colony areas, with vertical dashed lines marking the cut-off values for debris.
Influence of negative pressure on Staphylococcus aureus growth at different negative pressure intervals. This composite figure presents a series of box-and-whisker plots alongside density plots demonstrating the effects of varying levels of negative pressure on the growth characteristics of Staphylococcus aureus using different modes of intermittent negative pressure intervals. The first column of box plots in each panel depicts the colony-forming units (CFU), and the second illustrates the total growth area, across an intermittent negative pressure intervals of 1–3 h and the second illustrates the total growth area across intermittent negative pressure intervals of 1–3 h. The pressure used was 250 mmHg. The pressure used was 250 mmHg. The accompanying density plots visualize the distribution of colony areas, with vertical dashed lines marking the cut-off values for debris.

Inverse Gaussian multiple regression analysis for the growth of Staphylococcus aureus and Staphylococcus epidermidis regarding negative pressure in an in vitro wound model_

Dependent variable: CFU/ml
Pressure 9.21-15 (4.19-15)p = 0.0356
Species (S. epidermidis as reference) -3.06-12 (8.13-13)p = 0.0007
Pressure: species interaction -7.85-15 (4.27-15)p = 0.0761
Constant 3.55-12 (7.96-13)p = 0.0001
Observations 35
Log Likelihood -502.335
Akaike Inf. Crit. 1,012.670
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