scholarly journals A Wound-Healing Assay Based on Ultraviolet Light Ablation

2016 ◽  
Vol 22 (1) ◽  
pp. 36-43 ◽  
Author(s):  
Shang-Ying Wu ◽  
Yung-Shin Sun ◽  
Kuan-Chen Cheng ◽  
Kai-Yin Lo
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Healing Process ◽  
Cell Monolayer ◽  
Wound Healing Process ◽  
Collective Cell Migration ◽  
Wound Healing Assay ◽  
Physiological Processes ◽  
Free Region ◽  
A Cell

Collective cell migration plays important roles in many physiological processes such as embryonic development, tissue repair, and angiogenesis. A “wound” occurs when epithelial cells are lost and/or damaged due to some external factors, and collective cell migration takes place in the following wound-healing process. To study this cellular behavior, various kinds of wound-healing assays are developed. In these assays, a “wound,” or a “cell-free region,” is created in a cell monolayer mechanically, chemically, optically, or electrically. These assays are useful tools in studying the effects of certain physical or chemical stimuli on the wound-healing process. Most of these methods have disadvantages such as creating wounds of different sizes or shapes, yielding batch-to-batch variation, and damaging the coating of the cell culture surface. In this study, we used ultraviolet (UV) lights to selectively kill cells and create a wound out of a cell monolayer. A comparison between the current assay and the traditional scratch assay was made, indicating that these two methods resulted in similar wound-healing rates. The advantages of this UV-created wound-healing assay include fast and easy procedure, high throughput, and no direct contact to cells.

scholarly journals Effects of Substrate-Coating Materials on the Wound-Healing Process

Materials ◽  
2019 ◽  
Vol 12 (17) ◽  
pp. 2775 ◽  
Author(s):  
Jin-Young Lin ◽  
Kai-Yin Lo ◽  
Yung-Shin Sun
Keyword(s):  
Wound Healing ◽  
Healing Process ◽  
Petri Dish ◽  
Wound Healing Process ◽  
Collective Cell Migration ◽  
Dependent Manner ◽  
Wound Healing Assay ◽  
Coating Materials ◽  
Coated Surfaces ◽  
Physical And Chemical

The wound-healing assay is commonly and widely used for investigating collective cell migration under various physical and chemical stimuli. Substrate-coating materials are shown to affect the wound-healing process in a cell-type dependent manner. However, experiment-to-experiment variations make it difficult to compare results from different assays. In this paper, a modified barrier wound-healing assay was reported for studying the wound-healing process on different substrates in one single petri dish. In short, half of a dish was covered with the tape, and coating materials, poly-l-lysine and gelatin, were applied to the surface. After peeling off the tape, half of the surface was coated with the desired material. Then a customized barrier was placed inside the dish to create the wound. The results indicated that surface coating did not affect cell proliferation/viability, and the wound-healing rate increased in coated surfaces compared to uncoated ones. The present study provides a platform for further understanding the mechanisms of substrate coating-dependent wound-healing processes.

scholarly journals A microfluidics-based wound-healing assay for studying the effects of shear stresses, wound widths, and chemicals on the wound-healing process

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Jin-Young Lin ◽  
Kai-Yin Lo ◽  
Yung-Shin Sun
Keyword(s):  
Wound Healing ◽  
Shear Stress ◽  
Healing Process ◽  
Wound Healing Process ◽  
Shear Stresses ◽  
Collective Cell Migration ◽  
Wound Healing Assay ◽  
Normal Medium ◽  
Accelerate Wound Healing ◽  
Percent Area

AbstractCollective cell migration plays important roles in various physiological processes. To investigate this collective cellular movement, various wound-healing assays have been developed. In these assays, a “wound” is created mechanically, chemically, optically, or electrically out of a cellular monolayer. Most of these assays are subject to drawbacks of run-to-run variations in wound size/shape and damages to cells/substrate. Moreover, in all these assays, cells are cultured in open, static (non-circulating) environments. In this study, we reported a microfluidics-based wound-healing assay by using the trypsin flow-focusing technique. Fibroblasts were first cultured inside this chip to a cellular monolayer. Then three parallel fluidic flows (containing normal medium and trypsin solution) were introduced into the channels, and cells exposed to protease trypsin were enzymatically detached from the surface. Wounds of three different widths were generated, and subsequent wound-healing processes were observed. This assay is capable of creating three or more wounds of different widths for investigating the effects of various physical and chemical stimuli on wound-healing speeds. The effects of shear stresses, wound widths, and β-lapachone (a wound healing-promoting chemical) on wound-healing speeds were studied. It was found that the wound-healing speed (total area healed per unit time) increased with increasing shear stress and wound width, but under a shear stress of 0.174 mPa the linear healing speed (percent area healed per unit time) was independent of the wound width. Also, the addition of β-lapachone up to 0.5 μM did not accelerate wound healing. This microfluidics-based assay can definitely help in understanding the mechanisms of the wound-healing process and developing new wound-healing therapies.

AN OVERVIEW: INVESTIGATION OF ELECTROPORATION TECHNIQUE ON CELL PROPERTIES CULTURED ON MICROPATTERNED SURFACE.

2015 ◽  
Vol 77 (6) ◽  
Author(s):  
Nur Adilah Abd Rahman ◽  
Mamman Hassan Buhari ◽  
M. Mahadi Abdul Jamil
Keyword(s):  
Wound Healing ◽  
Cell Function ◽  
Microcontact Printing ◽  
Healing Process ◽  
Wound Healing Process ◽  
Medical Applications ◽  
Electrical Fields ◽  
A Cell ◽  
Basic Concepts ◽  
A New Technique

Electroporation (EP) is a method of controlling cell function by using pulses of electrical fields to create pore through a cell membrane and causes other substance around it to be absorbed into the cell. Where This method had been led to variety of medical applications. While, microcontact printing (μCP) is a quite useful technique for patterning extracellular matrix as an adhesion molecule for cells that works for controlling the cell growth. This study focuses on reviewing the basic concepts and techniques of electroporation and Microcontact printing, as applied to molecular biology & cancer treatment. The combination of these two technique might be a new technique for wound healing process treatment.

Platelet-Rich Plasma (PRP) Promotes Fetal Mesenchymal Stem/Stromal Cell Migration and Wound Healing Process

2014 ◽  
Vol 10 (3) ◽  
pp. 417-428 ◽  
Author(s):  
Maria G. Roubelakis ◽  
Ourania Trohatou ◽  
Apostolos Roubelakis ◽  
Evgenia Mili ◽  
Ioannis Kalaitzopoulos ◽  
...  
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Stromal Cell ◽  
Platelet Rich Plasma ◽  
Healing Process ◽  
Wound Healing Process

scholarly journals Collective migration during a gap closure in a two-dimensional haptotactic model

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Marie Versaevel ◽  
Laura Alaimo ◽  
Valentine Seveau ◽  
Marine Luciano ◽  
Danahe Mohammed ◽  
...  
Keyword(s):  
Cell Migration ◽  
Cell Density ◽  
Healing Process ◽  
Time Lapse ◽  
Wound Healing Process ◽  
Collective Cell Migration ◽  
Two Dimensional ◽  
Gap Closure ◽  
Lower Cell Density ◽  
Protein Gradients

AbstractThe ability of cells to respond to substrate-bound protein gradients is crucial for many physiological processes, such as immune response, neurogenesis and cancer cell migration. However, the difficulty to produce well-controlled protein gradients has long been a limitation to our understanding of collective cell migration in response to haptotaxis. Here we use a photopatterning technique to create circular, square and linear fibronectin (FN) gradients on two-dimensional (2D) culture substrates. We observed that epithelial cells spread preferentially on zones of higher FN density, creating rounded or elongated gaps within epithelial tissues over circular or linear FN gradients, respectively. Using time-lapse experiments, we demonstrated that the gap closure mechanism in a 2D haptotaxis model requires a significant increase of the leader cell area. In addition, we found that gap closures are slower on decreasing FN densities than on homogenous FN-coated substrate and that fresh closed gaps are characterized by a lower cell density. Interestingly, our results showed that cell proliferation increases in the closed gap region after maturation to restore the cell density, but that cell–cell adhesive junctions remain weaker in scarred epithelial zones. Taken together, our findings provide a better understanding of the wound healing process over protein gradients, which are reminiscent of haptotaxis.

scholarly journals Autologous micrograft accelerates endogenous wound healing response through ERK-induced cell migration

2019 ◽  
Vol 27 (5) ◽  
pp. 1520-1538 ◽  
Author(s):  
Martina Balli ◽  
Francesca Vitali ◽  
Adrian Janiszewski ◽  
Ellen Caluwé ◽  
Alvaro Cortés-Calabuig ◽  
...  
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Molecular Mechanism ◽  
Healing Process ◽  
Skin Lesions ◽  
Wound Healing Process ◽  
Transcriptional Signature ◽  
Beneficial Effects ◽  
Healing Response ◽  
Wound Healing Response

Abstract Defective cell migration causes delayed wound healing (WH) and chronic skin lesions. Autologous micrograft (AMG) therapies have recently emerged as a new effective and affordable treatment able to improve wound healing capacity. However, the precise molecular mechanism through which AMG exhibits its beneficial effects remains unrevealed. Herein we show that AMG improves skin re-epithelialization by accelerating the migration of fibroblasts and keratinocytes. More specifically, AMG-treated wounds showed improvement of indispensable events associated with successful wound healing such as granulation tissue formation, organized collagen content, and newly formed blood vessels. We demonstrate that AMG is enriched with a pool of WH-associated growth factors that may provide the starting signal for a faster endogenous wound healing response. This work links the increased cell migration rate to the activation of the extracellular signal-regulated kinase (ERK) signaling pathway, which is followed by an increase in matrix metalloproteinase expression and their extracellular enzymatic activity. Overall we reveal the AMG-mediated wound healing transcriptional signature and shed light on the AMG molecular mechanism supporting its potential to trigger a highly improved wound healing process. In this way, we present a framework for future improvements in AMG therapy for skin tissue regeneration applications.

scholarly journals Research Techniques Made Simple: Analysis of Collective Cell Migration Using the Wound Healing Assay

2017 ◽  
Vol 137 (2) ◽  
pp. e11-e16 ◽  
Author(s):  
Ayman Grada ◽  
Marta Otero-Vinas ◽  
Francisco Prieto-Castrillo ◽  
Zaidal Obagi ◽  
Vincent Falanga
Keyword(s):  
Wound Healing ◽  
Cell Migration ◽  
Collective Cell Migration ◽  
Wound Healing Assay ◽  
Simple Analysis ◽  
Research Techniques

scholarly journals Accurate Region-of-Interest Recovery Improves the Measurement of the Cell Migration Rate in the In Vitro Wound Healing Assay

2017 ◽  
Vol 22 (6) ◽  
pp. 626-635
Author(s):  
Cesar Bedoya ◽  
Andrés Cardona ◽  
July Galeano ◽  
Fabián Cortés-Mancera ◽  
Patrick Sandoz ◽  
...  
Keyword(s):  
Wound Healing ◽  
Cell Monolayer ◽  
Region Of Interest ◽  
Wound Healing Assay ◽  
Time Intervals ◽  
Positioning Errors ◽  
Cellular Events ◽  
Confluent Cell ◽  
A Cell

The wound healing assay is widely used for the quantitative analysis of highly regulated cellular events. In this essay, a wound is voluntarily produced on a confluent cell monolayer, and then the rate of wound reduction (WR) is characterized by processing images of the same regions of interest (ROIs) recorded at different time intervals. In this method, sharp-image ROI recovery is indispensable to compensate for displacements of the cell cultures due either to the exploration of multiple sites of the same culture or to transfers from the microscope stage to a cell incubator. ROI recovery is usually done manually and, despite a low-magnification microscope objective is generally used (10x), repositioning imperfections constitute a major source of errors detrimental to the WR measurement accuracy. We address this ROI recovery issue by using pseudoperiodic patterns fixed onto the cell culture dishes, allowing the easy localization of ROIs and the accurate quantification of positioning errors. The method is applied to a tumor-derived cell line, and the WR rates are measured by means of two different image processing software. Sharp ROI recovery based on the proposed method is found to improve significantly the accuracy of the WR measurement and the positioning under the microscope.

scholarly journals Method development and characterization of the low molecular weight peptidome of human wound fluids

2020 ◽  
Author(s):  
Mariena J.A. van der Plas ◽  
Jun Cai ◽  
Jitka Petrlova ◽  
Karim Saleh ◽  
Sven Kjellström ◽  
...  
Keyword(s):  
Wound Healing ◽  
Method Development ◽  
Healing Process ◽  
Unmet Need ◽  
Wound Healing Process ◽  
Promising Tool ◽  
Physiological Processes ◽  
Post Surgery ◽  
Surgical Wounds ◽  
Novel Biomarkers

AbstractWound infections are significant challenges globally, and there is an unmet need for better diagnosis of wound healing status and infection. The wound healing process is characterized by proteolytic events that are the result of basic physiological processes involving coagulation and complement activation, but also dysfunctional activations by endogenous and bacterial proteases. Peptides, downstream reporters of these proteolytic actions, could therefore serve as a promising tool for diagnosis of wound healing and infection. In the present study, we demonstrate a method for the characterisation of the complete peptidome of human wound fluids. We compare acute non-infected wound fluids obtained post-surgery with plasma samples and find significantly higher protein and peptide numbers in wound fluids, which typically were also smaller in size as compared to plasma-derived peptides. Finally, we analyse wound fluids collected from dressings after facial skin graft surgery and compare three uninfected and normally healing surgical wounds with three inflamed and S. aureus infected wounds. The results identify unique peptide patterns of various selected proteins including coagulation and complement factors, as well as proteases and antiproteinases. Together, the work defines a workflow for analysis of peptides derived from human wound fluids and demonstrate a proof-of-concept that such wound fluids can be used for analysis of qualitative differences of peptide patterns derived from wound fluids on larger patient cohorts, providing novel biomarkers for wound healing and infection.

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