Negative pressure accelerated monolayer keratinocyte healing involves Cdc42 mediated cell podia formation

Area of Science:

• Cell biology
• Wound healing research
• Biomedical engineering

Background:

• Negative-pressure wound therapy (NPWT) is a clinical technique for wound healing.
• The precise molecular mechanisms underlying NPWT's efficacy remain largely undefined.

Purpose of the Study:

• To investigate the role and localization of cell division control protein 42 (Cdc42) in keratinocytes under various negative pressures.
• To elucidate the time-course of Cdc42 pathway activation during NPWT.

Main Methods:

• Culturing keratinocytes in a negative pressure incubator at different pressures (AP, NP75, NP125, NP175).
• Evaluating wound closure time, cell morphology, and proliferation markers (PCNA).
• Analyzing Cdc42 and related protein expression in cell membranes, including experiments with Cdc42 knockdown.

Main Results:

• Optimal wound closure and cell podia formation were observed at NP125.
• Cdc42, N-WASP, and actin expression significantly increased in plasma membranes at NP125 after 12 hours.
• Cdc42 knockdown inhibited its expression at the cell leading edge.

Conclusions:

• Cdc42 pathway activation and localization are crucial for cell podia formation in keratinocytes under NPWT.
• NPWT likely accelerates wound healing by promoting cell migration through Cdc42-mediated mechanisms.