The wound healing assay revisited: A transport phenomena approach

Ascione, Flora; Caserta, Sergio; Guido, Stefano

doi:10.1016/j.ces.2016.11.014

Cited by 23 publications

(31 citation statements)

References 55 publications

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“…Since the evolution of cell speed was comparable between the wound healing and the randomly migrating hdFbs, we concluded that the highly directional migration is the key mediator of early dermal wound closure. This finding is in agreement with recent publications by Wickert et al and Ascione et al, which showed the rate of epithelial wound healing to be strongly correlated to directionality [40,47]. Furthermore, both studies also found the contribution of cell proliferation toward wound healing to be minimal.…”

Section: Resultssupporting

confidence: 93%

“…The relative contibution proliferation and migration on wound recovery can be estimated based on Thiele’s modulus [40,47], which is the ratio obtained by dividing the time scale of proliferation by the time scale of migration. Given the long duplication time of more than 48 h and a wound closure time of 33 h in our wound healing samples, the Thiele’s modulus is at least ~1.4.…”

Section: Resultsmentioning

confidence: 99%

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Extended live-tracking and quantitative characterization of wound healing and cell migration with SiR-Hoechst

Chung

Bellefeuille

Miller

et al. 2018

Experimental Cell Research

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Cell migration is essential to many life processes, including immune response, tissue repair, and cancer progression. A reliable quantitative characterization of the cell migration can therefore aid in the high throughput screening of drug efficacy in wound healing and cancer treatments. In this work, we report what we believe is the first use of SiR-Hoechst for extended live tracking and automated analysis of cell migration and wound healing. We showed through rigorous statistical comparisons that this far-red label does not affect migratory behavior. We observed excellent automated tracking of random cell migration, in which the motility parameters (speed, displacement, path length, directionality ratio, persistence time, and direction autocorrelation) obtained closely match those obtained from manual tracking. We also present an analysis framework to characterize the healing of a scratch wound from the perspective of single cells. The use of SiR-Hoechst is advantageous for the crowded environments in wound healing assays because as long as cell nuclei do not overlap, continuous tracking can be maintained even if there is cell-cell contact. In this paper, we report wound recovery based on the number of cells migrating into the wound over time, normalized by the initial cell count prior to the infliction of the wound. This normalized cell count approach is impervious to operator bias during the arbitration of wound edges and is also robust against variability that arises due to differences in the cell density of different samples. Additional wound healing characteristics were also defined based on the evolution of cell speed and directionality during healing. Not unexpected, the wound healing cells exhibited much higher tendency to maintain the same migratory direction in comparison to the randomly migrating cells. The use of SiR-Hoechst thus greatly simplified the automation of single cell and whole population analysis with high spatial and temporal resolution over extended periods of time.

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Section: Resultssupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Extended live-tracking and quantitative characterization of wound healing and cell migration with SiR-Hoechst

Chung

Bellefeuille

Miller

et al. 2018

Experimental Cell Research

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“…The total polyphenolic content of apple extract was assessed by Folin-Ciocalteu phenol reagent as described by Singleton et al 65 time-lapse video microscopy. Wound closure was tracked by automated time-lapse video microscopy 68,69 based on an inverted microscope (Zeiss Axiovert 200, Carl Zeiss, Jena Germany) enclosed in a homemade incubator that allows keeping the sample at a constant temperature (37 ± 0.1 °C), 5% CO 2 and 100% humidified atmosphere. Several independent fields of view in each cell dish were acquired by a high-resolution highsensitivity monochromatic CCD video camera (Hamamatsu Orca AG, Japan) using a long working distance 5 × objective in phase-contrast (CP Achromat Ph1), at regular intervals (20 min).…”

Section: Discussionmentioning

confidence: 99%

“…After an initial lag time (t l ), the obtained profiles, all starting from A/A 0 = 1, exhibited a linear trend, in agreement with the prediction of the Fisher-Kolmogoroff model 68 , suggesting that the wound healing process occurs at a constant velocity. The slope of the linear range of the A/A 0 vs. t curve can be considered a measure of the wound closure velocity α (h −1 ) 69 . In Supplementary Information a typical experimental curve where A/A 0 is plotted as a function of time for a control and a treated sample is described and graphically analyzed (Fig.…”

Section: Discussionmentioning

confidence: 99%

Annurca apple polyphenol extract promotes mesenchymal-to-epithelial transition and inhibits migration in triple-negative breast cancer cells through ROS/JNK signaling

Vuoso

D’Angelo

Ferraro

et al. 2020

Sci Rep

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Aberrant activation of epithelial-to-mesenchymal transition has been shown to correlate with triple-negative breast cancer (TNBC) progression and metastasis. Thus, the induction of the reverse process might offer promising opportunities to restrain TNBC metastatic spreading and related mortality. Recently, the Annurca apple polyphenol extract (APE) has been highlighted as a multi-faceted agent that selectively kills TNBC cells by ROS generation and sustained JNK activation. Here, by qualitatively and quantitatively monitoring the real-time movements of live cells we provided the first evidence that APE inhibited the migration of MDA-MB-231 and MDA-MB-468 TNBC cells and downregulated metalloproteinase-2 and metalloproteinase-9. In MDA-MB-231 cells APE decreased SMAD-2/3 and p-SMAD-2/3 levels, increased E-cadherin/N-cadherin protein ratio, induced the switch from N-cadherin to E-cadherin expression and greatly reduced vimentin levels. Confocal and scanning electron microscopy imaging of APE-treated MDA-MB-231 cells evidenced a significant cytoskeletal vimentin and filamentous actin reorganization and revealed considerable changes in cell morphology highlighting an evident transition from the mesenchymal to epithelial phenotype with decreased migratory features. Notably, all these events were reverted by N-acetyl-l-cysteine and JNK inhibitor SP600125 furnishing evidence that APE exerted its effects through the activation of ROS/JNK signaling. The overall data highlighted APE as a potential preventing agent for TNBC metastasis.

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“…Among various types of in vitro experiments, wound-healing assays are widely used to study cell migration and cell proliferation under different conditions including the use of different surface coatings (Ascione et al, 2017;Liberio et al, 2014;Tremel et al, 2009). Wound-healing assays are initiated by creating a scratch in a monolayer of cells, followed by the observation of how the remaining cells migrate and proliferate to close the gap over time.…”

Section: Introductionmentioning

confidence: 99%

Quantifying the role of different surface coatings in experimental models of wound healing

Wang

Sun

et al. 2020

Chemical Engineering Science

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In vitro surface coatings are widely used to mimic the role of extracellular matrix in the in vivo environment. Different effects are reported for different surface coatings, however, some of these results are inconsistent across the literature. To explore the role of different surface coatings, we use a new modified stopper-based wound-healing assay, called a stopper assay, with two commonly used surface coatings: gelatin and poly-L-lysine (PLL). Our experimental data show the gap width decreases faster with the gelatin and PLL coatings. Similarly, the number of cells in certain subregions increases faster with these coatings. Unfortunately, neither of these observations provides definitive mechanistic insight into the role of the coatings. To provide such insight we calibrate the solution of the Fisher-Kolmogorov model to match the experimental data. Our parameter estimates indicate that both coatings significantly increase cell motility without affecting cell proliferation.

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The wound healing assay revisited: A transport phenomena approach

Cited by 23 publications

References 55 publications

Extended live-tracking and quantitative characterization of wound healing and cell migration with SiR-Hoechst

Extended live-tracking and quantitative characterization of wound healing and cell migration with SiR-Hoechst

Annurca apple polyphenol extract promotes mesenchymal-to-epithelial transition and inhibits migration in triple-negative breast cancer cells through ROS/JNK signaling

Quantifying the role of different surface coatings in experimental models of wound healing

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