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

Hsu, Chih‐Chin; Chow, Shu‐Er; Chen, Carl Pai-Chu; Tsai, Wen–Chung; Yu, Shin-Ying; Lee, Sheng-Chi

doi:10.1016/j.jdermsci.2013.03.007

Cited by 22 publications

(20 citation statements)

References 32 publications

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“…In particular, NPWT has been shown to promote the healing rates and prevent wound infections by multiple mechanisms, including decreasing edema, removal of wound exudates, and translating physical stimulation to signal transduction in cells [26, 27]. Previous studies indicated that negative pressure conditions caused by NPWT could alter the gene expression and the function of host cells in vitro, such as bone marrow MSCs and keratinocytes [19, 20, 28]. However, its potential effects on P. aeruginosa and virulence factors have not been studied yet.…”

Section: Discussionmentioning

confidence: 99%

Effect of Negative Pressure on Proliferation, Virulence Factor Secretion, Biofilm Formation, and Virulence-Regulated Gene Expression ofPseudomonas aeruginosaIn Vitro

Wang

et al. 2016

BioMed Research International

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Objective. To investigate the effect of negative pressure conditions induced by NPWT on P. aeruginosa. Methods. P. aeruginosa was cultured in a Luria–Bertani medium at negative pressure of −125 mmHg for 24 h in the experimental group and at atmospheric pressure in the control group. The diameters of the colonies of P. aeruginosa were measured after 24 h. ELISA kit, orcinol method, and elastin-Congo red assay were used to quantify the virulence factors. Biofilm formation was observed by staining with Alexa Fluor® 647 conjugate of concanavalin A (Con A). Virulence-regulated genes were determined by quantitative RT-PCR. Results. As compared with the control group, growth of P. aeruginosa was inhibited by negative pressure. The colony size under negative pressure was significantly smaller in the experimental group than that in the controls (p < 0.01). Besides, reductions in the total amount of virulence factors were observed in the negative pressure group, including exotoxin A, rhamnolipid, and elastase. RT-PCR results revealed a significant inhibition in the expression level of virulence-regulated genes. Conclusion. Negative pressure could significantly inhibit the growth of P. aeruginosa. It led to a decrease in the virulence factor secretion, biofilm formation, and a reduction in the expression level of virulence-regulated genes.

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Section: Discussionmentioning

confidence: 99%

Effect of Negative Pressure on Proliferation, Virulence Factor Secretion, Biofilm Formation, and Virulence-Regulated Gene Expression ofPseudomonas aeruginosaIn Vitro

Wang

et al. 2016

BioMed Research International

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“…Activation and localization of Cdc42 proteins on cell membrane are involved in the cell podia formation in keratinocytes, which may facilitate cell migration to accelerate wound healing [46]. Thus, keratinocyte migration requires spatiotemporal integration of signaling molecules that regulate cell body motility.…”

Section: Discussionmentioning

confidence: 99%

β1 integrin signaling in asymmetric migration of keratinocytes under mechanical stretch in a co-cultured wound repair model

et al. 2016

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BackgroundKeratinocyte (KC) migration in re-epithelization is crucial in repairing injured skin. But the mechanisms of how mechanical stimuli regulate the migration of keratinocytes have been poorly understood.MethodsHuman immortalized keratinocyte HaCaT cells were co-cultured with skin fibroblasts on PDMS membranes and transferred to the static stretch device developed in-house for additional 6 day culture under mechanical stretch to mimic surface tension in skin. To detect the expression of proteins on different position at different time points and the effect of β1 integrin mechanotransduction on HaCaT migration, Immunofluorescence, Reverse transcription-polymerase chain reaction, Flow cytometry, Western blotting assays were applied.ResultsMechanical receptor of β1 integrin that recognizes its ligand of collagen I was found to be strongly associated with migration of HaCaT cells since the knockdown of β1 integrin via RNA silence eliminated the key protein expression dynamically. Here the expression of vinculin was lower but that of Cdc42 was higher for the cells at outward edge than those at inward edge, respectively, supporting that the migration capability of keratinocytes is inversely correlated with the formation of focal adhesion complexes but positively related to the lamellipodia formation. This asymmetric expression feature was further confirmed by high or low expression of PI3K for outward- or inward-migrating cells. And ERK1/2 phosphorylation was up-regulated by mechanical stretch.ConclusionWe reported here, a novel mechanotransduction signaling pathways were β1 integrin-dependent pattern of keratinocytes migration under static stretch in an in vitro co-culture model. These results provided an insight into underlying molecular mechanisms of keratinocyte migration under mechanical stimuli.

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“…Furthermore, application of NPWT can result in a significant rise of local vascular endothelial growth factor expression in both acute and chronic wounds, and NPWT also results in increased expression of platelet-derived growth factor and transforming growth factor-b1 in a vitro fibroblast model [30]. The activation of cell division control protein 42 (Cdc42) in the cell membrane is involved in the cell podia formation in keratinocytes, negative pressure at 125 mm Hg stimulates the appearance of Cdc42 in the leading edge cell membrane and activates downstream proteins to facilitate actin polymerization [19]. Above all, although using the molecular mechanism of NPWT on wounds remains undefined, refinements to achieve the desired outcomes have been determined largely by our clinical observation and empirical use.…”

Section: Discussionmentioning

confidence: 99%

“…5). The drainage tube of VSD was connected to a microcomputer electric drainage device for obtaining constant negative pressure, and the negative pressure was set at 125 mm Hg according to clinical practice and experiment in vitro, which demonstrated that negative pressure at 125 mm Hg stimulated wound healing significantly [18,19]. As our Burn Center has made several operations of microskin autografts covered by VSD before, we found that the time interval between skin grafting and first postoperative change should be 5e7 d according to patients' systemic and local situation (Figs.…”

Section: Treatment Protocolmentioning

confidence: 99%

Using negative pressure wound therapy on microskin autograft wounds

Zhang

Qiu

et al. 2015

Journal of Surgical Research

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

Cited by 22 publications

References 32 publications

Effect of Negative Pressure on Proliferation, Virulence Factor Secretion, Biofilm Formation, and Virulence-Regulated Gene Expression ofPseudomonas aeruginosaIn Vitro

Effect of Negative Pressure on Proliferation, Virulence Factor Secretion, Biofilm Formation, and Virulence-Regulated Gene Expression ofPseudomonas aeruginosaIn Vitro

β1 integrin signaling in asymmetric migration of keratinocytes under mechanical stretch in a co-cultured wound repair model

Using negative pressure wound therapy on microskin autograft wounds

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