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      An image J plugin for the high throughput image analysis of in vitro scratch wound healing assays

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          Abstract

          In vitro scratch wound healing assay, a simple and low-cost technique that works along with other image analysis tools, is one of the most widely used 2D methods to determine the cellular migration and proliferation in processes such as regeneration and disease. There are open-source programs such as imageJ to analyze images of in vitro scratch wound healing assays, but these tools require manual tuning of various parameters, which is time-consuming and limits image throughput. For that reason, we developed an optimized plugin for imageJ to automatically recognize the wound healing size, correct the average wound width by considering its inclination, and quantify other important parameters such as: area, wound area fraction, average wound width, and width deviation of the wound images obtained from a scratch/ wound healing assay. Our plugin is easy to install and can be used with different operating systems. It can be adapted to analyze both individual images and stacks. Additionally, it allows the analysis of images obtained from bright field, phase contrast, and fluorescence microscopes. In conclusion, this new imageJ plugin is a robust tool to automatically standardize and facilitate quantification of different in vitro wound parameters with high accuracy compared with other tools and manual identification.

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          TScratch: a novel and simple software tool for automated analysis of monolayer wound healing assays.

          Cell migration plays a major role in development, physiology, and disease, and is frequently evaluated in vitro by the monolayer wound healing assay. The assay analysis, however, is a time-consuming task that is often performed manually. In order to accelerate this analysis, we have developed TScratch, a new, freely available image analysis technique and associated software tool that uses the fast discrete curvelet transform to automate the measurement of the area occupied by cells in the images. This tool helps to significantly reduce the time needed for analysis and enables objective and reproducible quantification of assays. The software also offers a graphical user interface which allows easy inspection of analysis results and, if desired, manual modification of analysis parameters. The automated analysis was validated by comparing its results with manual-analysis results for a range of different cell lines. The comparisons demonstrate a close agreement for the vast majority of images that were examined and indicate that the present computational tool can reproduce statistically significant results in experiments with well-known cell migration inhibitors and enhancers.
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            Cancer-associated fibroblasts promote directional cancer cell migration by aligning fibronectin

            Cancer-associated fibroblasts (CAFs) in the tumor stroma play a key role in tumor progression. Erdogan et al. show that CAF-mediated alignment of the fibronectin matrix is a key factor promoting directional cancer cell migration.
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              Paracrine Effect of Mesenchymal Stem Cells Derived from Human Adipose Tissue in Bone Regeneration

              Mesenchymal stem cell (MSC) transplantation has proved to be a promising strategy in cell therapy and regenerative medicine. Although their mechanism of action is not completely clear, it has been suggested that their therapeutic activity may be mediated by a paracrine effect. The main goal of this study was to evaluate by radiographic, morphometric and histological analysis the ability of mesenchymal stem cells derived from human adipose tissue (Ad-MSC) and their conditioned medium (CM), to repair surgical bone lesions using an in vivo model (rabbit mandibles). The results demonstrated that both, Ad-MSC and CM, induce bone regeneration in surgically created lesions in rabbit's jaws, suggesting that Ad-MSC improve the process of bone regeneration mainly by releasing paracrine factors. The evidence of the paracrine effect of MSC on bone regeneration has a major impact on regenerative medicine, and the use of their CM can address some issues and difficulties related to cell transplants. In particular, CM can be easily stored and transported, and is easier to handle by medical personnel during clinical procedures.
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                Author and article information

                Contributors
                Role: Formal analysisRole: InvestigationRole: MethodologyRole: ValidationRole: Writing – original draft
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: Software
                Role: Methodology
                Role: MethodologyRole: SupervisionRole: Validation
                Role: MethodologyRole: SupervisionRole: ValidationRole: Writing – review & editing
                Role: Data curationRole: SupervisionRole: Writing – review & editing
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                28 July 2020
                2020
                : 15
                : 7
                : e0232565
                Affiliations
                [001]Department of Biomedical Engineering, Universidad de los Andes, Bogotá, Colombia
                Universita degli Studi di Milano-Bicocca, ITALY
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Author information
                http://orcid.org/0000-0001-6747-548X
                http://orcid.org/0000-0001-6238-9021
                Article
                PONE-D-20-11043
                10.1371/journal.pone.0232565
                7386569
                32722676
                af5ff77d-999b-4fd6-abfa-d0cf132121d3
                © 2020 Suarez-Arnedo et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 16 April 2020
                : 11 July 2020
                Page count
                Figures: 6, Tables: 0, Pages: 14
                Funding
                This work was financially supported by the Department of Biomedical Engineering at Universidad de los Andes, by the start-up funding to Assistant Professors FAPA (Fondo de Apoyo a Profesores Asistentes of Carolina Muñoz), and by an internal call for termination of publications (CI-001-Pública tus nuevos conocimientos o expone tus nuevas creaciones) from Vice-provost of Research at Universidad de los Andes.
                Categories
                Research Article
                Biology and Life Sciences
                Physiology
                Physiological Processes
                Tissue Repair
                Wound Healing
                Research and Analysis Methods
                Imaging Techniques
                Image Analysis
                Medicine and Health Sciences
                Diagnostic Medicine
                Diagnostic Radiology
                Magnetic Resonance Imaging
                Research and Analysis Methods
                Imaging Techniques
                Diagnostic Radiology
                Magnetic Resonance Imaging
                Medicine and Health Sciences
                Radiology and Imaging
                Diagnostic Radiology
                Magnetic Resonance Imaging
                Biology and Life Sciences
                Cell Biology
                Cell Motility
                Cell Migration
                Biology and Life Sciences
                Developmental Biology
                Cell Migration
                Engineering and Technology
                Equipment
                Laboratory Equipment
                Pipettes
                Research and Analysis Methods
                Imaging Techniques
                Physical Sciences
                Mathematics
                Geometry
                Radii
                Computer and Information Sciences
                Software Engineering
                Computer Software
                Open Source Software
                Engineering and Technology
                Software Engineering
                Computer Software
                Open Source Software
                Science Policy
                Open Science
                Open Source Software
                Custom metadata
                All relevant data are within the manuscript and its Supporting Information files.

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                Uncategorized

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