Scratch Wound Healing Assay

Author(s):  
Simona Martinotti ◽  
Elia Ranzato
2020 ◽  
Author(s):  
Alejandra Suarez-Arnedo ◽  
Felipe Torres Figueroa ◽  
Camila Clavijo ◽  
Pablo Arbeláez ◽  
Juan C. Cruz ◽  
...  

AbstractIn 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.


2021 ◽  
Vol 36 (1) ◽  
pp. 226-233
Author(s):  
Rukiye Boran ◽  
Nurdan Sarac ◽  
Tuba Baygar ◽  
Aysel Ugur

The genus Hypericum sp. has a number of uses in traditional medicine like curing the burns, ulcers, haemorrhoids and wound healing. The species Hypericum lydium Boiss. (Hypericaceae), however, has not been known to have any properties related to the healing of injuries or antimicrobial working against the oral microorganisms. The present study was aimed to evaluate the efficiency of H. lydium in soft tissue healing and its capacity to prevent infections after dental extraction. H. lydium was extracted with ethanol and the obtained extract was tested for its inhibition ability on extracellular matrix-degrading enzymes; collagenase, hyaluronidase and elastase. To determine the cytotoxicity and wound healing capacity of the extract, MTT and in vitro scratch wound healing assay were performed using the NIH-3T3 fibroblast cells, respectively. Antimicrobial activity was investigated by microdilution method against oral pathogenic microorganisms. The highest enzyme inhibition activity was determined against elastase (80.27±0.1%). According to the cytotoxic activity results, the IC50 value of the H. lydium was found to be 82.20±4.05 μg/mL. Scratch wound healing assay of the extract exhibited a significant enhancement at 24 h with a closed wound area when compared with the control. The extract showed potent antimicrobial properties against oral pathogenic microorganisms. The results of the study revealed out that H. lydium can be considered as a natural compound for dental industry to improve soft tissue healing and to prevent the possible infections after dental extraction.


Healthcare ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 911
Author(s):  
Entaz Bahar ◽  
Hyonok Yoon

The study of artificial neural networks (ANN) has undergone a tremendous revolution in recent years, boosted by deep learning tools. The presence of a greater number of learning tools and their applications, in particular, favors this revolution. However, there is a significant need to deal with the issue of implementing a systematic method during the development phase of the ANN to increase its performance. A multilayer feedforward neural network (FNN) was proposed in this paper to predict the cell migration assay on cisplatin-sensitive and cisplatin-resistant (CisR) ovarian cancer (OC) cell lines via scratch wound healing assay. An FNN training algorithm model was generated using the MATLAB fitting function in a MATLAB script to accomplish this task. The input parameters were types of cell lines, times, and wound area, and outputs were relative wound area, percentage of wound closure, and wound healing speed. In addition, we tested and compared the initial accuracy of various supervised learning classifier and support vector regression (SVR) algorithms. The proposed ANN model achieved good agreement with the experimental data and minimized error between the estimated and experimental values. The conclusions drawn demonstrate that the developed ANN model is a useful, accurate, fast, and inexpensive method to predict cancerous cell migration characteristics evaluated via scratch wound healing assay.


2009 ◽  
Vol 66 (12) ◽  
pp. 1041-1047 ◽  
Author(s):  
Manoj B. Menon ◽  
Natalia Ronkina ◽  
Jessica Schwermann ◽  
Alexey Kotlyarov ◽  
Matthias Gaestel

2019 ◽  
Vol 21 (Supplement_3) ◽  
pp. iii36-iii37
Author(s):  
M Ndiaye ◽  
C Rébé ◽  
A Ilie ◽  
L Ménégaut ◽  
T Pilot ◽  
...  

Abstract BACKGROUND Glioblastoma is the most common primary brain tumor. Its prognosis remains poor even with the standard treatment - the Stupp protocol.The classic Warburg effect in cancers leads to increased glycolysis which causes acidification of the tumor environment. This phenomenon may favor migration of tumor cells as already reported in pancreatic ductal adenocarcinoma. We therefore hypothesized that enhanced glycolysis in glioblastomas could favor the tumor cell migration. MATERIAL AND METHODS We measured glycolysis by the extracellular acidification rate (ECAR) of several human glioblastomas cell lines (LN229, LN18, T98-G, U87-MG, U373-MG, U118-MG) with the Seahorse Analyzer. To confirm these results, we also measured the intracellular cAMP rates using the Cayman’s Elisa kit and we analyzed by RT-PCR the expression of the main genes coding for enzymes involved in glycolysis in these glioblastomas cell lines. Cell migration was measured with a scratch wound healing assay during 24 hours. RESULTS U118-MG was the glioblastoma cell line with the highest glycolysis rate, the highest production of cAMP and showed a strong expression of glycolysis-associated genes. LN229 was the glioblastoma cell line with the less important glycolysis rate, the lower production of cAMP and showed a weaker expression of glycolysis-associated genes. According to the scratch wound healing assay, U118-MG cells showed a more important migration than LN229 cells at 24 hours. CONCLUSION Glycolysis may be an attractive target to prevent effectively tumor cell migration in glioblastomas. Coupling the evaluation of glycolysis with histomolecular characterization of glioblastomas, could help to identify patients to whom adjuvant therapies that inhibit glycolysis such as fenofibrate could be proposed.


Pharmaceutics ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 2195
Author(s):  
Angela Fabiano ◽  
Chiara Migone ◽  
Luca Cerri ◽  
Anna Maria Piras ◽  
Andrea Mezzetta ◽  
...  

Olive leaves extract (OLE) has been extensively studied as antioxidant and antibiotic and these characteristics make it particularly interesting for use on wounds. For this reason, the aim of this study was to introduce OLE in microparticles (MP) of hyaluronic acid (MPHA-OLE) or chitosan (MPCs-OLE) to obtain a spray patch for the treatment of wounds in anatomical areas that are difficult to protect with traditional patches. The MP were characterized for particle size and ability to protect OLE from degradation, to absorb water from wound exudate, to control OLE release from MP. The MPHA and MPCs medicated or not and mixtures of the two types in different proportions were studied in vitro on fibroblasts by the scratch wound healing assay. The MP size was always less than 5 µm, and therefore, suitable for a spray patch. The MPCs-OLE could slow down the release of OLE therefore only about 60% of the polyphenols contained in it were released after 4 h. Both MPHA and MPCs could accelerate wound healing. A 50% MPHA-OLE-50% MPCs-OLE blend was the most suitable for accelerating wound healing. The MPHA-OLE-MPCs-OLE blends studied in this work were shown to have the characteristics suitable for a spray patch, thus giving a second life to the waste products of olive growers.


2016 ◽  
Vol 38 (6) ◽  
pp. 2152-2162 ◽  
Author(s):  
Lei Lin ◽  
Hong-bin Tu ◽  
Liang Wu ◽  
Ming Liu ◽  
Ge-ning Jiang

Background/Aims: SMAD7 is a key inhibitor of transforming growth factor β (TGFβ) receptor signaling, which regulates the alteration of cancer cell invasiveness through epithelial-mesenchymal cell conversion. Carboplatin is a commonly used drug in the chemotherapy for non-small cell lung cancer (NSCLC). Nevertheless, the molecular mechanisms underlying its suppressive effects on the NSCLC cell invasion are not completely understood. In the current study, we addressed this question by analyzing the effects of Carboplatin on microRNA-regulated SMAD7. Methods: We used Carboplatin to treat NSCLC cell lines. We performed bioinformatics analyses on the binding of microRNA-21 (miR-21) to the 3'-UTR of SMAD7 mRNA, and verified the biological effects of this binding using promoter luciferase reporter assay. The effects of Carboplatin or miR-21-modification on NSCLC cell invasion were evaluated in either a transwell cell invasion assay, or a scratch wound healing assay. Results: We found that Carboplatin inhibited the NSCLC cell invasion, in either a transwell cell invasion assay, or a scratch wound healing assay. Moreover, Carboplatin increased the levels of SMAD7 protein, but not mRNA, in NSCLC cells, suggesting presence of post-transcriptional control of SMAD7 by Carboplatin. Furthermore, expression of miR-21 was found to be inhibited by Carboplatin, and bioinformatics analyses showed that miR-21 targeted the 3'-UTR of SMAD7 mRNA to inhibit its translation, which was confirmed by luciferase reporter assay. Conclusion: Carboplatin may upregulate SMAD7 through suppression of miR-21 to inhibit TGFβ receptor signaling mediated NSCLC cell invasion.


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