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

Polish journal of microbiology Pub Date : 2024-06-20 eCollection Date: 2024-06-01 DOI:10.33073/pjm-2024-018
Adam Bobkiewicz, Wojciech Francuzik, Amy Martinkosky, Maciej Borejsza-Wysocki, Witold Ledwosinski, Krzysztof Szmyt, Tomasz Banasiewicz, Lukasz Krokowicz
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Abstract

Negative Pressure Wound Therapy (NPWT) has been widely adopted in wound healing strategies due to its multimodal mechanism of action. While NPWT's positive impression on wound healing is well-established, its effect on bacterial load reduction remains equivocal. This study investigates NPWT's efficacy in reducing bioburden using an in vitro porcine skin model, focusing on the impact of Staphylococcus aureus and Staphylococcus epidermidis. Custom-made negative pressure chambers were employed to apply varying negative pressures. Porcine skin was cut into 5 × 5 cm squares and three standardized wounds of 6 mm each were created using a biopsy punch. Then, wounds were infected with S. aureus and S. epidermidis bacterial suspensions diluted 1:10,000 to obtain a final concentration of 1.5 × 104 CFU/ml and were placed in negative pressure chambers. After incubation, bacterial counts were expressed as colony-forming units (CFU) per ml. For S. aureus at 120 hours, the median CFU, mean area per colony, and total growth area were notably lower at -80 mmHg when compared to -250 mmHg and -50 mmHg, suggesting an optimal negative pressure for the pressure-dependent inhibition of the bacterial proliferation. While analyzing S. epidermidis at 120 hours, the response to the negative pressure was similar but less clear, with the minor CFU at -100 mmHg. The influence of intermittent negative pressure on the S. epidermidis growth showed notably lower median CFU with the interval therapy every hour compared to the S. aureus control group. This study contributes valuable insights into NPWT's influence on the bacterial load, emphasizing the need for further research to reformulate its role in managing contaminated wounds.

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负压水平及其对体外伤口模型中细菌生长动力学的影响
负压伤口疗法(NPWT)因其多模式的作用机制而在伤口愈合策略中被广泛采用。虽然 NPWT 对伤口愈合的积极作用已得到证实,但其对减少细菌负荷的效果仍不明确。本研究使用体外猪皮肤模型研究了 NPWT 在减少生物负载方面的功效,重点关注金黄色葡萄球菌和表皮葡萄球菌的影响。采用定制的负压室来施加不同的负压。猪皮肤被切割成 5 × 5 厘米的正方形,并用活检打孔器造成三个各 6 毫米的标准化伤口。然后,将金黄色葡萄球菌和表皮葡萄球菌细菌悬浮液按 1:10,000 的比例稀释至最终浓度为 1.5 × 104 CFU/ml,并将其置于负压室中。培养后,细菌计数以每毫升菌落形成单位(CFU)表示。对于 120 小时的金黄色葡萄球菌,与 -250 mmHg 和 -50 mmHg 相比,-80 mmHg 下的 CFU 中位数、每个菌落的平均面积和总生长面积都明显较低,这表明压力依赖性抑制细菌增殖的最佳负压值。在分析 120 小时的表皮葡萄球菌时,对负压的反应类似,但不太明显,在-100 mmHg 时的 CFU 较少。间歇负压对表皮葡萄球菌生长的影响显示,与金黄色葡萄球菌对照组相比,每隔一小时治疗一次,表皮葡萄球菌的 CFU 中位数明显降低。这项研究为了解 NPWT 对细菌负荷的影响提供了宝贵的见解,强调了进一步研究其在管理污染伤口中的作用的必要性。
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