Related Experiment Video
Updated: Aug 12, 2025
Development of a Benchtop Model for Evaluating the Compatibility of Wound Dressing Materials with Negative Pressure Wound Therapy Systems
Published on: May 2, 2025
Novel cell culture system for monitoring cells during continuous and variable negative-pressure wound therapy
Toshifumi Yamashiro1, Toshihiro Kushibiki2, Yoshine Mayumi2
1Department of Plastic and Reconstructive Surgery, National Defense Medical College, Tokorozawa, Saitama, Japan.
Background:
Although the clinical efficacy of negative-pressure wound therapy (NPWT) is well known, many of its molecular biological mechanisms remain unresolved, mainly due to the difficulty and paucity of relevant in vitro studies. We attempted to develop an in vitro cell culture system capable of real-time monitoring of cells during NPWT treatment.
Materials And Methods:
A novel negative-pressure cell culture system was developed by combining an inverted microscope, a stage-top incubator, a sealed metal chamber for cell culture, and an NPWT treatment device. Human keratinocytes, PSVK-1, were divided into ambient pressure (AP), continuous negative-pressure (NPc), and intermittent negative-pressure (NPi) groups and cultured for 24 h with scratch assay using our real-time monitoring system and device. Pressure inside the device, medium evaporation rate, and the residual wound area were compared across the groups.
Results:
Pressure in the device was maintained at almost the same value as set in all groups. Medium evaporation rate was significantly higher in the NPi group than in the other two groups; however, it had negligible effect on cell culture. Residual wound area after 9 h evaluated by the scratch assay was significantly smaller in the NPc and NPi groups than in the AP group.
Conclusion:
We developed a negative-pressure cell culture device that enables negative-pressure cell culture under conditions similar to those used in clinical practice and is able to monitor cells under NPWT. Further experiments using this device would provide high-quality molecular biological evidence for NPWT.
Insights
A new in vitro system allows real-time monitoring of cells during negative-pressure wound therapy (NPWT). This system demonstrated that NPWT accelerates wound healing in keratinocytes, providing a platform for further NPWT research.
Area of Science:
- Biomedical Engineering
- Cell Biology
- Wound Healing Research
Background:
- Clinical efficacy of negative-pressure wound therapy (NPWT) is established, but underlying molecular mechanisms are poorly understood.
- Existing in vitro studies are limited, hindering mechanistic investigations of NPWT.
Purpose of the Study:
- To develop an in vitro cell culture system for real-time monitoring of cellular responses to NPWT.
- To investigate the effects of continuous and intermittent negative pressure on keratinocyte wound closure.
Main Methods:
- A novel system integrated an inverted microscope, incubator, sealed chamber, and NPWT device.
- Human keratinocytes (PSVK-1) were cultured under ambient pressure (AP), continuous NPWT (NPc), and intermittent NPWT (NPi) for 24 hours.
- Scratch assays were used to evaluate residual wound area, with pressure and medium evaporation monitored.
Main Results:
- The NPWT system maintained stable negative pressure across groups.
- Intermittent NPWT (NPi) showed a higher medium evaporation rate, with negligible impact on cell culture.
- Significant reduction in residual wound area was observed in both continuous NPWT (NPc) and NPi groups compared to ambient pressure (AP) after 9 hours.
Conclusions:
- A functional in vitro negative-pressure cell culture device enabling real-time monitoring was successfully developed.
- This system mimics clinical NPWT conditions and facilitates cellular investigation.
- The device provides a valuable tool for generating molecular evidence to elucidate NPWT mechanisms.
