Journal articles: 'Scratch Assay'

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Alishahedani, Mohammadali E., Manoj Yadav, Katelyn J. McCann, Portia Gough, Carlos R. Castillo, Jobel Matriz, and Ian A. Myles. "Therapeutic candidates for keloid scars identified by qualitative review of scratch assay research for wound healing." PLOS ONE 16, no. 6 (June 18, 2021): e0253669. http://dx.doi.org/10.1371/journal.pone.0253669.

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The scratch assay is an in vitro technique used to analyze cell migration, proliferation, and cell-to-cell interaction. In the assay, cells are grown to confluence and then ‘scratched’ with a sterile instrument. For the cells in the leading edge, the resulting polarity induces migration and proliferation in attempt to ‘heal’ the modeled wound. Keloid scars are known to have an accelerated wound closure phenotype in the scratch assay, representing an overactivation of wound healing. We performed a qualitative review of the recent literature searching for inhibitors of scratch assay activity that were already available in topical formulations under the hypothesis that such compounds may offer therapeutic potential in keloid treatment. Although several shortcomings in the scratch assay literature were identified, caffeine and allicin successfully inhibited the scratch assay closure and inflammatory abnormalities in the commercially available keloid fibroblast cell line. Caffeine and allicin also impacted ATP production in keloid cells, most notably with inhibition of non-mitochondrial oxygen consumption. The traditional Chinese medicine, shikonin, was also successful in inhibiting scratch closure but displayed less dramatic impacts on metabolism. Together, our results partially summarize the strengths and limitations of current scratch assay literature and suggest clinical assessment of the therapeutic potential for these identified compounds against keloid scars may be warranted.

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Vargas, Andrea, Marc Angeli, Chiara Pastrello, Rosanne McQuaid, Han Li, Andrea Jurisicova, and Igor Jurisica. "Robust quantitative scratch assay." Bioinformatics 32, no. 9 (December 31, 2015): 1439–40. http://dx.doi.org/10.1093/bioinformatics/btv746.

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Gough, Wendy, Keren I. Hulkower, Renee Lynch, Patrick Mcglynn, Mark Uhlik, Lei Yan, and Jonathan A. Lee. "A Quantitative, Facile, and High-Throughput Image-Based Cell Migration Method Is a Robust Alternative to the Scratch Assay." Journal of Biomolecular Screening 16, no. 2 (February 2011): 155–63. http://dx.doi.org/10.1177/1087057110393340.

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Cell migration is a key phenotype for a number of therapeutically important biological responses, including angiogenesis. A commonly used method to assess cell migration is the scratch assay, which measures the movement of cells into a wound made by physically scoring a confluent cell monolayer to create an area devoid of cells. Although this method has been adequate for qualitative characterization of migration inhibitors, it does not provide the highly reproducible results required for quantitative compound structure-activity relationship evaluation because of the inconsistent size and placement of the wound area within the microplate well. The Oris™ Cell Migration Assay presents a superior alternative to the scratch assay, permitting formation of precisely placed and homogeneously sized cell-free areas into which migration can occur without releasing factors from wounded or dead cells or damaging the underlying extracellular matrix. Herein the authors compare results from the scratch and Oris™ cell migration assays using an endothelial progenitor cell line and the Src kinase inhibitor dasatinib. They find that using the Acumen™ Explorer laser microplate cytometer in combination with the Oris™ Cell Migration Assay plate provides a robust, efficient, and cost-effective cell migration assay exhibiting excellent signal to noise, plate uniformity, and statistical validation metrics.

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Bobadilla, Ana Victoria Ponce, Jazmine Arévalo, Eduard Sarró, Helen M. Byrne, Philip K. Maini, Thomas Carraro, Simone Balocco, Anna Meseguer, and Tomás Alarcón. "In vitro cell migration quantification method for scratch assays." Journal of The Royal Society Interface 16, no. 151 (February 2019): 20180709. http://dx.doi.org/10.1098/rsif.2018.0709.

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The scratch assay is an in vitro technique used to assess the contribution of molecular and cellular mechanisms to cell migration. The assay can also be used to evaluate therapeutic compounds before clinical use. Current quantification methods of scratch assays deal poorly with irregular cell-free areas and crooked leading edges which are features typically present in the experimental data. We introduce a new migration quantification method, called ‘monolayer edge velocimetry’, that permits analysis of low-quality experimental data and better statistical classification of migration rates than standard quantification methods. The new method relies on quantifying the horizontal component of the cell monolayer velocity across the leading edge. By performing a classification test on in silico data, we show that the method exhibits significantly lower statistical errors than standard methods. When applied to in vitro data, our method outperforms standard methods by detecting differences in the migration rates between different cell groups that the other methods could not detect. Application of this new method will enable quantification of migration rates from in vitro scratch assay data that cannot be analysed using existing methods.

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Gotsulyak, N. Ya, V. R. Kosach, O. V. Cherednyk, I. O. Tykhonkova, and A. I. Khoruzhenko. "Optimization of cell motility evaluation in scratch assay." Biopolymers and Cell 30, no. 3 (May 20, 2014): 223–28. http://dx.doi.org/10.7124/bc.00089d.

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Putri, Widya Wira, Nadya Muthia Risky, Nungki Pramita Sari, Rizka Kurnia Gemilang, Rufaida Mudrika, Indra Kusuma, Restu Syamsul Hadi, and Yurika Sandra. "Dimethyl Sulfoxide Inhibits CD271+ Migration in Scratch Assay." Advanced Science Letters 23, no. 7 (July 1, 2017): 6881–83. http://dx.doi.org/10.1166/asl.2017.9423.

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Jana, Snehasis, and Mahendra Kumar Trivedi. "Wound Healing Activity of Consciousness Energy Healing Treatmenton HFF-1 Cells and DMEM Using Scratch Assay." Investigative Dermatology and Venereology Research 4, no. 1 (December 26, 2018): 50–54. http://dx.doi.org/10.15436/2381-0858.18.2036.

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The wound healing activity using scratch assay is considered as a convenient in vitro tool for the assessment of wound healing. The present study deals with the optimization of Biofield Energy Treatment (Consciousness Energy Healing Treatment-The Trivedi Effect®) in the HFF-1 cell line (Human Foreskin Fibroblast) and DMEM (Dulbecco's Modified Eagle Medium) using scratch assay against positive control, recombinant Human Epidermal Growth Factor (Hu EGF, 30 ng/mL). This methodwas used for the determination of cell proliferation and migration of fibroblast quantitatively in the scratched wounded area. The scratched area was monitored after 24 hours of wound closure in the Biofield Energy Treated HFF-1 cells and the Biofield Energy Treated DMEM groups, and the representative photomicrographs were taken in each wells using WimScratch Image analysis software. The results showed that the Biofield Energy Treated DMEMwassignificantly higher percentage of fibroblast migration i.e. 51.8%, while the migration was altered in the Biofield Energy Treated HFF-1 cell line compared to the baseline control group. In addition to, the percentage of scratch area was significantly decreased by 2.7% in the Biofield Energy Treated DMEM group, while it was increased by 12% in the Biofield Treated cells group compared to the baseline control group. Overall, the experimental results concluded that The Trivedi Effect® has the significant capacity and wide implications in wound healing activityvia cell culture media, DMEM as compared with the HFF-1 cell line directly. Biofield Energy Healing would be a complementary and alternative medicine that can be used against burn injury cases, acute wound, skin regeneration, eczema, diaper rash, chickenpox, measles, warts, acne, hives, wrinkles, ringworm, Rosacea, psoriasis, seborrheic dermatitis, skin cancer, etc.

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Minadakis, Georgios, Emmanouil Angelakis, Dimosthenis Chochlakis, Yannis Tselentis, and Anna Psaroulaki. "Cat-Scratch disease in Crete: an update." Infectious Disease Reports 3, no. 2 (December 5, 2011): 15. http://dx.doi.org/10.4081/idr.2011.3210.

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There are few epidemiological and clinical studies about the presence of cat scratch disease (CSD) on the island of Crete. The objective of this study was to analyze a large number of patients with suspected CSD to define the frequency of Bartonella infections in Crete. From January 2005 to October 2008, we studied patients with suspected CSD from hospitals in Crete. Sera of the referred patients were tested by immunofluorescence assay (IFA). For some patients, we also received lymph nodes and blood samples that we tested for the presence of Bartonella henselae by molecular assays. Overall, we tested 507 serum samples and we found 56 (11%) cases of CSD. PCR assay was positive for 2 patients; one had a B. henselae positive lymph node and the other a positive whole blood sample. Significantly more CSD cases (62.5%, 35 of 56) were reported in children than in infants and adults (P<0.05). Moreover, we identified that most cases of CSD occurred between May and September (P=0.002) and December and January. CSD is prevalent in Crete and is mostly associated with an increase in outdoor activity.

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Liu, Lili, Zhiying Xu, Binbin Yu, Li Tao, and Ying Cao. "Berbamine Inhibits Cell Proliferation and Migration and Induces Cell Death of Lung Cancer Cells via Regulating c-Maf, PI3K/Akt, and MDM2-P53 Pathways." Evidence-Based Complementary and Alternative Medicine 2021 (July 8, 2021): 1–20. http://dx.doi.org/10.1155/2021/5517143.

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Berbamine (BBM) is a natural product isolated from Berberis amurensis Rupr. We investigated the influence of BBM on the cell viability, proliferation, and migration of lung cancer cells and explored the possible mechanisms. The cell viability and proliferation of lung cancer cells were evaluated by MTT assay, EdU assay, and colony formation assay. Migration and invasion abilities of cancer cells were determined through wound scratch assay and Transwell assay. Cell death was evaluated by cell death staining assay and ELISA. The expressions of proteins were evaluated using western blot assay. A xenograft mouse model derived from non-small-cell lung cancer cells was used to detect the effect of BBM on tumor growth and metastasis in vivo. Both colony formation and EdU assays results revealed that BBM (10 μM) significantly inhibited the proliferation of A549 cells ( P < 0.001 ). BBM (10 μM) also significantly inhibited the migration and invasion ability of cancer cells in wound scratch and Transwell assays. Trypan blue assay and ELISA revealed that BBM (20 μM) significantly induced cell death of A549 cells. In xenograft mouse models, the tumor volume was significantly smaller in mice treated with BBM (20 mg/kg). The western blotting assay showed that BBM inhibited the PI3K/Akt and MDM2-p53 signaling pathways, and BBM downregulated the expression of c-Maf. Our results show that BBM inhibits proliferation and metastasis and induces cell death of lung cancer cells in vitro and in vivo. These effects may be achieved by BBM reducing the expression of c-Maf and regulating the PI3K/Akt and MDM2-p53 pathways.

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Scholz, H. M., I. Aykara, K. Frommer, S. Rehart, U. Müller-Ladner, and E. Neumann. "AB0044 ENDOTHELIAL CELL AND RHEUMATOID ARTHRITIS SYNOVIAL FIBROBLAST MIGRATION AND ADHESION ARE ALTERED BY ACTIVIN/FOLLISTATIN." Annals of the Rheumatic Diseases 80, Suppl 1 (May 19, 2021): 1054.2–1055. http://dx.doi.org/10.1136/annrheumdis-2021-eular.271.

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Background:Activin A and follistatin belong to an anti-inflammatory auto-regulatory cycle. Patients with rheumatoid arthritis (RA) have increased activin A levels in the synovial fluid and tissue. During inflammation, activin A is released systemically, then inducing its antagonist follistatin. This negative feedback is active in different cell types but not RA synovial fibroblasts (SF). Fibroblasts interact with endothelial cells in inflamed tissues inducing angiogenesis.Objectives:Evaluation of the role of activin A and follistatin on RASF and endothelial cell interactions.Methods:RA synovium was used for RASF isolation, HUVEC were commercially obtained. RASF and HUVEC were stimulated in mono- and coculture. Direct: RASF together with HUVEC; indirect: inserts with HUVEC separated by a membrane from RASF. Stimuli: activin A 15ng/ml, follistatin 500ng/ml, IL-1β 1ng/ml. Proliferation was analyzed by BrdU assay. RASF were Calcein-AM stained. Cells were transferred to 24-well plates after 18h stimulation. After adhesion for 1h, non-adherent cells were removed by shaking 3x for 5 min. Afterwards, fluorescent cells were quantified. For the flow-adhesion assay, HUVEC were cultured on rattail collagen coated capillary slides. HUVEC and RASF were stimulated for 4h with TNFα, TNFα+activin A or TNFα+follistatin. After stimulation, 2x10^6 RASF were resuspended in 20ml medium and sent through the capillaries. Two 1min videos were recorded (18.4ml/h, 30.5ml/h). Settings: TNFα-stimulated RASF on HUVEC stimulated wit TNFα or activin A+TNFα or follistatin+ TNFα. For migration assays, 2% FCS medium with 1x10^5 cells were placed in inserts (8µm membrane) into wells with 10% FCS (control: 2%FCS vs. 2%FCS) and stimulated with, IL-1, IL-1+activinA and IL-1+follistatin. After 16h, migrated cells were quantified. For scratch-assay 4x10^5 cells were cultured overnight, then cells were scratched and stimulated, afterwards 3 pictures per scratch were taken at start, after 10h and every 1.5h. Cells migrating into the scratch area were quantified.Results:IL-1 induced activin A in RASF and HUVEC in all settings. IL-1-induced activin A release was reduced by follistatin in HUVEC monoculture and both cocultures compared to IL-1 alone but not in RASF monoculture. IL-1-induced IL-6 release was reduced by activin A in HUVEC and indirect coculture but not in RASF monoculture and direct coculture. Follistatin did not alter IL-6 responses. IL-1 induced VEGF in RASF but not in HUVEC and was not altered by activin A. Short-term adhesion showed no significant influence of activin A or follistatin (n=4). Flow adhesion assay showed reduced adherence/rolling of RASF on HUVEC stimulated with TNFα and activin A (n=4). Migration assays showed that IL-1 decreased migration but without significant difference between the induced effects mediated by IL-1+activinA and IL-1+follistatin (n=4). Scratch assay showed increased migration in dicrect coculture with greater difference between stimulated and unstimulated cells in RASF monoculture and indirect coculture (n=4). Proliferation was not altered by activin A or follistatin.Conclusion:In direct and indirect coculture of HUVEC with RASF the effect on HUVEC was dominant leading to reduced IL-1-induced activin A release. However, the IL-1-induced IL-6 release in RASF or HUVECs was decreased by activin A in HUVEC monoculture and indirect coculture but not during cell-contact between both cell types. The direct interaction of RASF with HUVEC seems to prevent the reducing activin A effect on IL-6 release in HUVECs. Activin A seems to not to have an impact on short-term cell adhesion but first observations show, that activin A alters selectin-mediated adhesion under flow conditions. The migration assay shows that IL-1-induced effects on cell migration were enhanced by activin A and follistatin. Migration assay shows that IL-1-induced decrease on migration more prominent in indirect coculture and RASF monoculture than in direct coculture although in gap migration in the scratch assay was highest in direct coculture.Disclosure of Interests:None declared

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Paul, N. "Sudhangsu assay: a scratch of green chemistry for beginners." Journal of Physics: Conference Series 1913, no. 1 (May 1, 2021): 012086. http://dx.doi.org/10.1088/1742-6596/1913/1/012086.

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Radstake, Wilhelmina E., Kiran Gautam, Cynthia Van Rompay, Randy Vermeesen, Kevin Tabury, Mieke Verslegers, Sarah Baatout, and Bjorn Baselet. "Comparison of in vitro scratch wound assay experimental procedures." Biochemistry and Biophysics Reports 33 (March 2023): 101423. http://dx.doi.org/10.1016/j.bbrep.2023.101423.

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Félix, Rute Castelo, Liliana Anjos, Rita Alves Costa, Sophia Letsiou, and Deborah Mary Power. "Cartilage Acidic Protein a Novel Therapeutic Factor to Improve Skin Damage Repair?" Marine Drugs 19, no. 10 (September 25, 2021): 541. http://dx.doi.org/10.3390/md19100541.

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Fish skin has been gaining attention due to its efficacy as a human-wound-treatment product and to identify factors promoting its enhanced action. Skin fibroblasts have a central role in maintaining skin integrity and secrete extra cellular matrix (ECM) proteins, growth factors and cytokines to rapidly repair lesions and prevent further damage or infection. The effects on scratch repair of the ubiquitous but poorly characterized ECM protein, cartilage acidic protein 1 (CRTAC1), from piscine and human sources were compared using a zebrafish SJD.1 primary fibroblast cell line. A classic in vitro cell scratch assay, immunofluorescence, biosensor and gene expression analysis were used. Our results demonstrated that the duplicate sea bass Crtac1a and Crtac1b proteins and human CRTAC-1A all promoted SJD.1 primary fibroblast migration in a classic scratch assay and in an electric cell impedance sensing assay. The immunofluorescence analysis revealed that CRTAC1 enhanced cell migration was most likely caused by actin-driven cytoskeletal changes and the cellular transcriptional response was most affected in the early stage (6 h) of scratch repair. In summary, our results suggest that CRTAC1 may be an important factor in fish skin promoting damage repair.

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Saha, Susmita, Deepjyoti Bhattacharjee, Anwesha Saha, Gahin De, Partha Saha, and S. K. Sil. "Wound healing promoting activity of Earthworm, Eutyphoeus gammiei (Beddard): in vitro studies on human skin keratinocyte cell line (HaCat)." Journal of Drug Delivery and Therapeutics 8, no. 6 (November 15, 2018): 155–58. http://dx.doi.org/10.22270/jddt.v8i6.2036.

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Earthworm, Eutyphoeus gammiei, homogenate (EGH) was screened for wound healing activity on human keratinocyte cell line, HaCat, by cell proliferation and migration assays. The maximum proliferation and migration of keratinocyte cells were observed at the dose of 25μg/ml. As cell proliferation and migration are key factors for wound healing, the study clearly suggests the potential role of earthworm species Eutyphoeus gammiei on wound healing. Keywords: Eutyphoeus gammiei, Keratinocyte, MTT assay, scratch assay.

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Al-Amin, Mohammad, Nagla Mustafa Eltayeb, Chowdhury Faiz Hossain, Melati Khairuddean, Siti Sarah Fazalul Rahiman, and Salizawati Muhamad Salhimi. "Inhibitory Activity of Extract, Fractions, and Compounds from Zingiber montanum Rhizomes on the Migration of Breast Cancer Cells." Planta Medica 86, no. 06 (March 13, 2020): 387–94. http://dx.doi.org/10.1055/a-1129-7026.

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Abstract Zingiber montanum rhizomes are traditionally used for the treatment of numerous human ailments. The present study was carried out to investigate the inhibitory activity of the crude extract, chromatographic fractions, and purified compounds from Z. montanum rhizomes on the migration of MDA-MB-231 cells. The effect of the extract on cell migration was investigated by a scratch assay, which showed significant inhibition in a concentration-dependent manner. Vacuum liquid chromatography on silica gel afforded four fractions (Frs. 1 – 4), which were tested on cell migration in the scratch assay. Frs. 1 and 2 showed the most significant inhibition of MDA-MB-231 cell migration. The effect of the most potent fraction (Fr. 2) was further confirmed in a transwell migration assay. The study of Frs. 1 and 2 by gelatin zymography showed significant inhibition of MMP-9 enzyme activity. Chromatographic separation of Frs. 1 and 2 afforded buddledone A (1), zerumbone (2), (2E,9E)-6-methoxy-2,9-humuradien-8-one (3), zerumbone epoxide (4), stigmasterol (5), and daucosterol (6). In a cell viability assay, compounds 1 – 4 inhibited the viability of MDA-MB-231 cells in a concentration-dependent manner. The study of buddledone A (1) and zerumbone epoxide (4) on cell migration revealed that 4 significantly inhibited the migration of MDA-MB-231 cells in both scratch and transwell migration assays. The results of the present study may lead to further molecular studies behind the inhibitory activity of zerumbone epoxide (4) on cell migration and support the traditional use of Z. montanum rhizomes for the treatment of cancer.

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Freiesleben, Sara H., Jens Soelberg, Nils T. Nyberg, and Anna K. Jäger. "Determination of the Wound Healing Potentials of Medicinal Plants Historically Used in Ghana." Evidence-Based Complementary and Alternative Medicine 2017 (2017): 1–6. http://dx.doi.org/10.1155/2017/9480791.

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The present study was carried out to investigate the wound healing potentials of 17 medicinal plants historically used in Ghana for wound healing. Warm and cold water extracts were prepared from the 17 dried plant species and tested in vitro in the scratch assay with NIH 3T3 fibroblasts from mice. The wound healing scratch assay was used to evaluate the effect of the plants on cell proliferation and/or migration in vitro, as a test for potential wound healing properties. After 21 hours of incubation increased proliferation and/or migration of fibroblasts in the scratch assay was obtained for 5 out of the 17 plant species. HPLC separation of the most active plant extract, which was a warm water extract of Philenoptera cyanescens, revealed the wound healing activity to be attributed to rutin and a triglycoside of quercetin. The present study suggests that Allophylus spicatus, Philenoptera cyanescens, Melanthera scandens, Ocimum gratissimum, and Jasminum dichotomum have wound healing activity in vitro.

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Emrich, Stefanie, Anja Schuster, Thomas Schnabel, and Gertie Janneke Oostingh. "Antimicrobial Activity and Wound-Healing Capacity of Birch, Beech and Larch Bark Extracts." Molecules 27, no. 9 (April 28, 2022): 2817. http://dx.doi.org/10.3390/molecules27092817.

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Bark is a major by-product of woodworking industries. The contents of several wood species are known to harbor antimicrobial, antiviral, anti-inflammatory and wound-healing capacities. The aim of this work was to identify beneficial properties of Austrian larch, birch and beech bark extracts for their potential usage as additives or active ingredients in dermatological applications. Bacterial agar diffusion assay and resazurin-based broth microdilution assay were used to evaluate anti-bacterial activity. To gain more insight into the cellular response to bark extracts, viability-, scratch-assays and ELISAs were performed. Birch and beech extracts showed strong antimicrobial activities against Gram-positive bacteria, including Cutibacterium acnes, Staphylococcus epidermidis and MRSA. Wound closure was enhanced with birch and beech extracts as compared to controls in the scratch-assays. Whereas beneficial properties of birch bark components have previously been described, the similar effects of beech extracts are novel. The combined positive effect on wound-healing and antimicrobial activity has great potential for the treatment of various skin diseases, including acne in future dermal applications.

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Johnston, Stuart T., Matthew J. Simpson, and D. L. Sean McElwain. "How much information can be obtained from tracking the position of the leading edge in a scratch assay?" Journal of The Royal Society Interface 11, no. 97 (August 6, 2014): 20140325. http://dx.doi.org/10.1098/rsif.2014.0325.

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Moving cell fronts are an essential feature of wound healing, development and disease. The rate at which a cell front moves is driven, in part, by the cell motility, quantified in terms of the cell diffusivity D , and the cell proliferation rate λ . Scratch assays are a commonly reported procedure used to investigate the motion of cell fronts where an initial cell monolayer is scratched, and the motion of the front is monitored over a short period of time, often less than 24 h. The simplest way of quantifying a scratch assay is to monitor the progression of the leading edge. Use of leading edge data is very convenient because, unlike other methods, it is non-destructive and does not require labelling, tracking or counting individual cells among the population. In this work, we study short-time leading edge data in a scratch assay using a discrete mathematical model and automated image analysis with the aim of investigating whether such data allow us to reliably identify D and λ . Using a naive calibration approach where we simply scan the relevant region of the ( D , λ ) parameter space, we show that there are many choices of D and λ for which our model produces indistinguishable short-time leading edge data. Therefore, without due care, it is impossible to estimate D and λ from this kind of data. To address this, we present a modified approach accounting for the fact that cell motility occurs over a much shorter time scale than proliferation. Using this information, we divide the duration of the experiment into two periods, and we estimate D using data from the first period, whereas we estimate λ using data from the second period. We confirm the accuracy of our approach using in silico data and a new set of in vitro data, which shows that our method recovers estimates of D and λ that are consistent with previously reported values except that that our approach is fast, inexpensive, non-destructive and avoids the need for cell labelling and cell counting.

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Silva, Paulo Sérgio Gomes da, Raquel Ferreira Lopes, Jeferson Caetano Da Silva, Wanderlei Barbosa Dos Santos, Regina Célia Sales Santos Veríssimo, and Maria Lysete De Assis Bastos. "Cytotoxic, antimicrobial and healing activity of the Jatropha gossypiifolia L extract." Revista de Enfermagem UFPE on line 12, no. 2 (February 4, 2018): 465. http://dx.doi.org/10.5205/1981-8963-v12i2a234689p465-474-2018.

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RESUMOObjetivo: investigar o potencial citotóxico, antimicrobiano e cicatrizante de extratos das folhas, galhos e caule da J. gossypiifolia L. Método: estudo quantitativo, experimental. Os extratos foram obtidos por maceração em etanol, concentrados em evaporador rotatório e secos em dessecador a vácuo. Na análise, realizaram-se testes de prospecção fitoquímica; citotoxidade; microdiluição em caldo e Scratch assay. Entre os metabólitos detectados, estiveram: taninos, esteroides, flavonoides, flavonas, xantonas. Resultados: o extrato do caule apresentou viabilidade celular acima de 80%. As folhas foram moderadamente citotóxicas e os galhos apresentaram ausência de viabilidade celular. Os extratos inibiram o crescimento de S. aureus, S. epidermidis e P. aeruginosa em diferentes concentrações. O Scratch assay evidenciou que a fração metanólica das folhas propiciou a migração celular em 45% a mais do que o controle. Conclusão: estudos com esta espécie vegetal devem ser continuados para isolamento do princípio ativo, visando à produção de um fitoterápico cicatrizante de feridas. Descritores: Cicatrização; Enfermagem; Citoxidade; Plantas Medicinais.ABSTRACTObjective: to investigate the cytotoxic, antimicrobial and cicatrizant potential of extracts of leaves, branches and stem of J. gossypiifolia L. Method: quantitative, experimental study. The extracts were obtained by maceration in ethanol, concentrated in a rotary evaporator and dried in a vacuum desiccator. In the analysis, phytochemical prospecting; cytotoxicity; microdilution in broth and Scratch assay tests were performed. Among the detected metabolites, were: tannins, steroids, flavonoids, flavones, xanthones. Results: the stem extract presented cell viability above 80%. The leaves were moderately cytotoxic and the branches showed no cell viability. The extracts inhibited the growth of S. aureus, S. epidermidis and P. aeruginosa at different concentrations. Scratch assay showed that the methanolic fraction of the leaves allowed the cellular migration in 45% more than the control. Conclusion: studies with this plant species should be continued for isolation of the active principle, aiming at the production of a wound healing phytotherapic. Descriptors: Healing; Nursing; Citotoxicity; Medicinal Plants.RESUMENObjetivo: investigar el potencial citotóxico, antimicrobiano y cicatrizante de los extractos de las hojas, ramas y el tallo de J. gossypiifolia L. Método: estudio cuantitativo, experimental. Los extractos fueron obtenidos por maceración en etanol, concentrados en evaporador rotatorio y seco en desecador al vacuo. En el análisis, se realizaron pruebas de prospección fitoquímica; citotoxicidad; microdilución en caldo y Scratch asay. Entre los metabolitos detectados, estuvieron: taninos, esteroides, flavonoides, flavonas, xantonas. Resultados: el extracto del tallo presentó viabilidad celular por encima del 80%. Las hojas fueron moderadamente citotóxicas y las ramas presentaron ausencia de viabilidad celular. Los extractos inhibieron el crecimiento de S. aureus, S. epidermidis y P. aeruginosa en diferentes concentraciones. El Scratch assay evidenció que la fracción metanólica de las hojas propició la migración celular en un 45% más que el control. Conclusión: estudios con esta especie vegetal deben ser continuados para aislamiento del principio activo, visando la producción de un fitoterápico cicatrizante de heridas. Descriptores: La Curación; Enfermería; Citotoxicidad; Plantas Medicinales.

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Zhang, Tie-ning, Quan Li, Te Ba, Tian-xi Shao, Fang Li, and Ling-feng Wang. "Effects of platelet-rich plasma on in vitro proliferation and migration of fibroblasts from human chronic refractory wound granulation tissue." Discussion of Clinical Cases 8, no. 1 (August 18, 2021): 24. http://dx.doi.org/10.5430/dcc.v8n1p24.

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Objective: To observe the effects of platelet-rich plasma (PRP) on in vitro proliferation and migration of fibroblasts from human chronic refractory wound granulation tissue.Methods: Fibroblasts were separated from human chronic refractory wound granulation tissue and then were identified. The obtained fibroblasts were divided into fetal bovine serum (FBS) group, hydrogel group and PRP group, and the three groups were cultured with culture mediums containing FBS, hydrogel and PRP respectively, in order to observe the growth of fibroblasts. The wound scratch assay was used to observe the migration of fibroblasts.Results: PRP group had more fibroblasts than FBS group and hydrogel group since Day 5 of culture, and exhibited greater fibroblast scratch migration area than FBS group on 48 h and 72 h of wound scratch assay (all p < .05).Conclusions: Compared with FBS, human fibroblasts cultured by PRP can more effectively promote the proliferation and migration of fibroblasts.

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Salac, Edcyl Lee O., Michael Russelle Alvarez, Rnie Shayne Gaurana, Sheryl Joyce B. Grijaldo, Luster Mae Serrano, Florence de Juan, Rowell Abogado, et al. "Biological Assay-Guided Fractionation and Mass Spectrometry-Based Metabolite Profiling of Annona muricata L. Cytotoxic Compounds against Lung Cancer A549 Cell Line." Plants 11, no. 18 (September 12, 2022): 2380. http://dx.doi.org/10.3390/plants11182380.

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Annona muricata L. (Guyabano) leaves are reported to exhibit anticancer activity against cancer cells. In this study, the ethyl acetate extract from guyabano leaves was purified through column chromatography, and the cytotoxic effects of the semi-purified fractions were evaluated against A549 lung cancer cells using in vitro MTS cytotoxicity and scratch/wound healing assays. Fractions F15-16C and F15-16D exhibited the highest anticancer activity in the MTS assay, with % cytotoxicity values of 99.6% and 99.4%, respectively. The bioactivity of the fractions was also consistent with the results of the scratch/wound healing assay. Moreover, untargeted metabolomics was employed on the semi-purified fractions to determine the putative compounds responsible for the bioactivity. The active fractions were processed using LC-MS/MS analysis with the integration of the following metabolomic tools: MS-DIAL (for data processing), MetaboAnalyst (for data analysis), GNPS (for metabolite annotation), and Cytoscape (for network visualization). Results revealed that the putative compounds with a significant difference between active and inactive fractions in PCA and OPLS-DA models were pheophorbide A and diphenylcyclopropenone.

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Parabucki, Ana, Anja Santrač, Danijela Savić, Sanja Dacić, Ivana Bjelobaba, Sanja Peković, and Mirjana Stojiljković. "Real-Time PCR and Immunocytochemical Study of Chondroitin Sulfate Proteoglycans after Scratch Wounding in Cultured Astrocytes / PCR I IMUNOCITOHEMIJSKA STUDIJA EKSPRESIJE HONDROITIN-SULFATNIH PROTEOGLIKANA NAKON POVREDE ASTROCITA U KULTURI." Journal of Medical Biochemistry 32, no. 4 (October 1, 2013): 398–405. http://dx.doi.org/10.2478/jomb-2013-0036.

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Summary Background: Various in vivo and in vitro models have been described in order to elucidate the pathobiology underlying the traumatic brain injury (TBI) and test potentially suitable treatments. Since TBI is a complex disease, models differ in regard to the aspect of TBI that is being investigated. One of the used in vitro models is the scratch wound assay, first established as a reproducible, low-cost assay for the analysis of cell migration in vitro. The aim of the present study was to further investigate the relevancy of this model as a counter- part of in vivo TBI models. Methods: We have examined the astrocytic response to a mechanical injury in terms of expression of chondroitin sul- fate proteoglycans (CSPGs) - phosphacan, neurocan and brevican, using real-time PCR and immunocytochemistry. Results: Our results indicate that in vitro scratch wounding alters the expression profile of examined CSPGs. Four hours after the scratch injury of the astrocytic monolayer, real-time PCR analysis revealed upregulation of mRNA levels for phos- phacan (3-fold) and neurocan (2-fold), whereas brevican mRNA was downregulated (2-fold). Immunofluorescent sig- nal for phosphacan and neurocan was more intense in astro- cytes close to the injury site, while brevican was scarcely present in cultured astrocytes. Conclusions: Obtained results indicate that CSPGs are differ- entially expressed by astrocytes after scratch wounding, demonstrating that the scratch wound model might be suit- able for investigation of astrocyte-derived response to injury.

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Abbas, ManalAhmad, ManalMohammad Abbas, Naseer Al-Rawi, and Iqbal Al-Khateeb. "Naringenin potentiated β-sitosterol healing effect on the scratch wound assay." Research in Pharmaceutical Sciences 14, no. 6 (2019): 566. http://dx.doi.org/10.4103/1735-5362.272565.

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Liu, James X., Jordan Werner, Thorsten Kirsch, Joseph D. Zuckerman, and Mandeep S. Virk. "Cytotoxicity evaluation of chlorhexidine gluconate on human fibroblasts, myoblasts, and osteoblasts." Journal of Bone and Joint Infection 3, no. 4 (August 10, 2018): 165–72. http://dx.doi.org/10.7150/jbji.26355.

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Abstract. Introduction: Chlorhexidine gluconate (CHX) is widely used as a preoperative surgical skin-preparation solution and intra-wound irrigation agent, with excellent efficacy against wide variety of bacteria. The cytotoxic effect of CHX on local proliferating cells following orthopaedic procedures is largely undescribed. Our aim was to investigate the in vitro effects of CHX on primary fibroblasts, myoblasts, and osteoblasts.Methods: Cells were exposed to CHX dilutions (0%, 0.002%, 0.02%, 0.2%, and 2%) for either a 1, 2, or 3-minute duration. Cell survival was measured using a cytotoxicity assay (Cell Counting Kit-8). Cell migration was measured using a scratch assay: a “scratch” was made in a cell monolayer following CHX exposure, and time to closure of the scratch was measured.Results: All cells exposed to CHX dilutions of ≥ 0.02% for any exposure duration had cell survival rates of less than 6% relative to untreated controls (p < 0.001). Cells exposed to CHX dilution of 0.002% all had significantly lower survival rates relative to control (p < 0.01) with the exception of 1-minute exposure to fibroblasts, which showed 96.4% cell survival (p = 0.78). Scratch defect closure was seen in < 24 hours in all control conditions. However, cells exposed to CHX dilutions ≥ 0.02% had scratch defects that remained open indefinitely.Conclusions: The clinically used concentration of CHX (2%) permanently halts cell migration and significantly reduces survival of in vitro fibroblasts, myoblasts, and osteoblasts. Further in vivo studies are required to examine and optimize CHX safety and efficacy when applied near open incisions or intra-wound application.

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Ban, Weng Kit, Isabel Lim Fong, Heng Yen Khong, and Joyce Hui Yie Phung. "Wound Healing, Antimicrobial and Antioxidant Properties of Clinacanthus nutans (Burm.f.) Lindau and Strobilanthes crispus (L.) Blume Extracts." Molecules 27, no. 5 (March 6, 2022): 1722. http://dx.doi.org/10.3390/molecules27051722.

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Clinacanthus nutans is known to be an anticancer and antiviral agent, and Strobilanthes crispus has proven to be an antidiuretic and antidiabetic agent. However, there is a high possibility that these plants possess multiple beneficial properties, such as antimicrobial and wound healing properties. This study aims to assess the wound healing, antioxidant, and antimicrobial properties of Clinacanthus nutans and Strobilanthes crispus. The Clinacanthus nutans and Strobilanthes crispus leaves were dried, ground, and extracted with ethanol, acetone, and chloroform through cold maceration. In a modified scratch assay with co-incubation of skin fibroblast and Methicillin-resistant Staphylococcus aureus, Clinacanthus nutans and Strobilanthes crispus extracts were assessed for their wound healing potential, and the antimicrobial activities of Clinacanthus nutans and Strobilanthes crispus extracts were performed on a panel of Gram-positive and Gram-negative bacteria on Mueller–Hinton agar based on a disc diffusion assay. To assess for antioxidant potential, 2,2-diphenyl-1-picrylhydrazyl (DPPH), total phenolic and total flavonoid assays were conducted. In the modified scratch assay, Clinacanthus nutans extracts aided in the wound healing activity while in the presence of MRSA, and Strobilanthes crispus extracts were superior in antimicrobial and wound healing activities. In addition, Strobilanthes crispus extracts were superior to Clinacanthus nutans extracts against Pseudomonas aeruginosa on Mueller–Hinton agar. Acetone-extracted Clinacanthus nutans contained the highest level of antioxidant in comparison with other Clinacanthus nutans extracts.

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Liu, Haiwang, Ran Liu, Meiling Hao, Xing Zhao, and Chunhui Li. "Knockdown of KIF3C Inhibits Epithelial-Mesenchymal Transition in Lung Cancer Cells A549." Journal of Clinical and Nursing Research 6, no. 5 (September 26, 2022): 50–56. http://dx.doi.org/10.26689/jcnr.v6i5.4360.

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The objective of this study is to reveal the role of KIF3C gene in the proliferation of lung cancer cells, and the regulation of epithelial mesenchymal transition (EMT) of tumor cells. The plate clone formation assay and cell scratch assay were used in this study to detect the changes of cell proliferation and migration ability after siKIF3C interference, while EMT-related protein expression after KIF3C downregulation was detected by Western blot. The cell clone formation assay showed that the number of clones of lung cancer cells A549 was significantly reduced after transfected with siKIF3C (P<0.05); The scratch assay showed that the healing ability of cells was significantly reduced after transfected with siKIF3C (P<0.05); Western blot protein analysis revealed that the levels of EMT-related proteins, N-cadherin, Vimentin, Snail, and Slug were significantly down-regulated (P<0.05), however, E-cadherin protein levels were up-regulated after siKIF3C interference. In conclusion, KIF3C may promote the proliferation and invasive ability of lung cancer cells A549 through EMT.

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Xu, Bo, Qin Shao, Kaipeng Xie, Yuqing Zhang, Tianyu Dong, Yankai Xia, and Wei Tang. "The Long Non-Coding RNA ENST00000537266 and ENST00000426615 Influence Papillary Thyroid Cancer Cell Proliferation and Motility." Cellular Physiology and Biochemistry 38, no. 1 (2016): 368–78. http://dx.doi.org/10.1159/000438637.

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Background/Aims: Papillary thyroid cancer (PTC) is the most common histotype of Thyroid cancer (TC). Here, we detected the differentially expressed lncRNAs in tumor tissues and non-tumor tissues of PTC patients by lncRNA microarrays, and explored the function and molecular mechanisms of lncRNAs in the pathogenesis of PTC using a PTC cell line. Methods: CCK-8 assay, colony formation assay and EdU assay were used to detect the cell viability. Flow Cytometry was used to detect the cell cycle and apoptosis. Transwell and scratch assay were used to detect the cell motility. Results: CCK-8 assay, colony formation assay and EdU assay revealed that lncRNAs (ENST00000537266 and ENST00000426615) could inhibit cell proliferation. Cell cycle analysis showed that cell proportion was statistically significant increased in G1 phase and decreased in S phase and G2 phase in Si-266 transfected TPC-1 cells. In addition, a noteworthy increase of cell proportion in G1 phase accompanied by a decrease in S phase and unchanged G2 phase in Si-615 transfected TPC-1 cells were also observed. Meanwhile, transwell and scratch assay showed that ENST00000426615 could inhibit the cell motility while ENST00000537266 could not. Conclusion: Our results showed that lncRNAs (ENST00000426615 and ENST00000537266) might be important regulators of PTC cell proliferation and motility, which might provide new insight into the understanding of PTC pathogenesis.

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Hoey, John G., Fernando Valois-Cruz, Hannah Goldenberg, Yekaterina Voskoboynik, Jenna Pfiffner, Richard C. Tilton, Eli Mordechai, and Martin E. Adelson. "Development of an Immunoglobulin M Capture-Based Enzyme-Linked Immunosorbent Assay for Diagnosis of Acute Infections with Bartonella henselae." Clinical and Vaccine Immunology 16, no. 2 (December 3, 2008): 282–84. http://dx.doi.org/10.1128/cvi.00305-08.

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ABSTRACT We describe the development of an immunoglobulin M-specific enzyme-linked immunosorbent assay for the detection of an early antibody response to Bartonella henselae, the causative agent of cat scratch disease, bacillary angiomatosis, and endocarditis. This assay discriminates between B. henselae-positive and -negative patient samples with sensitivity and specificity values of 100% and 97.1%, respectively.

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Zhao, Shuangshuang, Junhui Zhao, and Ni Zhang. "Hyal1 Expression in Colorectal Carcinoma Cell Migration and Invasiveness: Significance and Mechanism." Evidence-Based Complementary and Alternative Medicine 2022 (July 5, 2022): 1–5. http://dx.doi.org/10.1155/2022/4418300.

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Objective. To clarify the significance of hyaluronidsase 1 (Hyal1) expression in colorectal carcinoma (CRC) and its impact on tumor cell migration and invasiveness. Methods. Human CRC cell lines SW480, HCT116, and SW620 were purchased, ELISA and western blot were used to detect the expression of Hyal1 in cells, CCK-8 assay to detect cell proliferation ability, cell scratch assay to check cell migration rate, and cell invasion was detected by the transwell assay. The correlation of Hyal1 with CRC cell migration and invasiveness capacities was analyzed. Result. ELISA results showed that supernatant Hyal1 level was the lowest in SW480, highest in HCT116, with the level in SW620 in between ( P < 0.05 ). No evident difference was identified by western blot in Hyal1 protein expression among the three cells ( P > 0.05 ). The cell scratch assay and transwell assay showed that the migration and invasion ability of HCT116 cells was higher than that of SW620 ( P < 0.05 ). In vitro, Hyal1 had a synergistic relationship with the invasiveness and migration capacities of CRC cells ( P < 0.05 ). Conclusion. Hyal1 is elevated in CRC and is consistent with the invasiveness and metastasis abilities of CRC cells. It is hoped that this research can provide reference for future prevention and treatment of CRC.

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Suriyah, Wastuti Hidayati, Aisyah Juares Rizal, Hana Syakirah Mohamed Nadzirin, Solachuddin Jauhari Arief Ichwan, and Muhammad Lokman Md Isa. "In Vitro Wound Healing Effect of Asiaticoside Extracted from Centella asiatica (‘Pegaga’) on Human Gingival Fibroblast Cell Line." Materials Science Forum 1025 (March 2021): 224–29. http://dx.doi.org/10.4028/www.scientific.net/msf.1025.224.

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Asiaticoside is a bioactive compound found in the traditional plant Centella asiatica (Asiatic pennywort or ‘Pegaga’) generally used for wound healing applications. Numerous studies have discussed the potential benefits of asiaticoside on different human cells such as keratinocytes and dermal fibroblast cells in healing of wounds. However only very few studies have been conducted to investigate its healing effect on cells originated from human oral cavity. The present study aimed to determine the potential of asiaticoside on human gingival fibroblast cells. Cytotoxic activities of the compounds were assessed by MTT assay. The wound healing was examined by scratch assay. The effect of asiaticoside on Col1A1 gene expression was also analyzed using qRT-PCR. Col1A1 is known to play a crucial role in wound healing. The MTT assay result showed that the maximum tolerable concentration of asiaticoside was 0.25 mg/ml. The scratch assay revealed that asiaticoside significantly accelerated the wound healing compared to the negative control (P<0.05). Moreover, the qRT-PCR demonstrated that asiaticoside markedly increased Col1A1 mRNA expression. These results proved asiaticoside as a potential candidate for wound healing agent in dentistry.

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İğci, Bahar, and Zeki Aytaç. "Phytochemical composition of Verbascum stachydifolium Boiss & Heldr. var. stachydifolium growing in Türkiye and in vitro analysis of wound healing activity." Archives of Biological Sciences, no. 00 (2023): 1. http://dx.doi.org/10.2298/abs221222001k.

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This study aimed to investigate the phenolic content, antioxidant activity, cytotoxicity and the in vitro wound healing activity of methanolic and aqueous extracts of Verbascum stachydifolium Boiss & Heldr. var. stachydifolium. Total phenolic and flavonoid contents and antioxidant activity were measured using spectrophotometry-based methods. Quantitative analysis of the selected phenolics was performed by HPLC. The cytotoxic effects of the extracts on L929 mouse fibroblast cells were evaluated by the MTT assay. The migration of treated fibroblast cells was assessed by the cell scratch assay. The expressions of type I collagen, FGF7, TGF-?1 and VEGF were evaluated by qRT-PCR and ELISA. The HPLC-based analysis revealed the presence of different phenolic compounds at varying amounts and high antioxidant activities were detected. The cytotoxicity assay results indicated that the methanolic and aqueous extracts did not exhibit any cytotoxic effect on fibroblast cells when used up to 500 ?g/mL concentration. Fibroblast migration was stimulated to the highest degree by the aqueous extract obtained by maceration as observed in the scratch assay at 60.4% closure. The molecular mechanism of the wound healing activity involves the upregulation of the analyzed genes.

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Sagala, Evayanti Meiliana, and Jansen Silalahi. "Wound Healing Activities of Hydrolyzed Virgin Coconut Oil (HVCO) and Fucoidan Combination: An In Vitro Assay." Asian Journal of Pharmaceutical Research and Development 7, no. 3 (June 14, 2019): 40–45. http://dx.doi.org/10.22270/ajprd.v7i3.532.

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combination, in the NIH 3T3 cell line using in vitro assay, and compared with single HVCO and single fucoidan. Methods: NIH 3T3 Cell viability and proliferation were assessed using the MTT method, migration activity was assessed using scratch wound healing assays and expression of COX-2 and VEGF protein were determined using immunocytochemistry (ICC). Results: The results from the proliferative activity assay show that the effective concentrations for all samples were 31.25 μg /ml. NIH 3T3 cells migration activity assay showed that the best combination of the HVCO and fucoidan was 50:50. From COX 2 and VEGF protein expression test results, the combination of HVCO and fucoidan has a higher percentage of expression than single HVCO or single fucoidan Conclusion: The results reveal that the combination of HVCO and fucoidan has better wound healing activity than single HVCO or single fucoidan

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Wu, Shang-Ying, Yung-Shin Sun, Kuan-Chen Cheng, and Kai-Yin Lo. "A Wound-Healing Assay Based on Ultraviolet Light Ablation." SLAS TECHNOLOGY: Translating Life Sciences Innovation 22, no. 1 (July 10, 2016): 36–43. http://dx.doi.org/10.1177/2211068216646741.

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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.

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de Lima, Tielidy A. de M., Gabriel Goetten de Lima, Bor Shin Chee, Jeferson G. Henn, Yvonne J. Cortese, Mailson Matos, Cristiane V. Helm, Washington L. E. Magalhães, and Michael J. D. Nugent. "Characterization of Gels and Films Produced from Pinhão Seed Coat Nanocellulose as a Potential Use for Wound Healing Dressings and Screening of Its Compounds towards Antitumour Effects." Polymers 14, no. 14 (July 7, 2022): 2776. http://dx.doi.org/10.3390/polym14142776.

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The reuse of agro-industrial waste assumes great importance today. Pinhão is the seed of Araucaria angustifolia, which is native to the mountains of southern Brazil, Paraguay, and Argentina. The coat is a by-product of this seed and is rich in phenolic compounds. The present study aimed to use the residue as a precursor material for the production of nanocellulose through the mechanical defibrillation process and perform the characterization of the films and the gel to investigate the effect on the physical and regenerative properties when incorporated with polyvinyl alcohol (PVA). The modulus of elasticity was higher when the MFC of pinhão was added to the PVA. Film and gel had their cytotoxicity tested by MTT assay using 3T3 fibroblast and Schwann cancer cells, and a migration assay was also performed using the scratch test on HaCat keratinocyte cells. For the scratch test, film and gel samples with low concentration presented a complete scratch closure in 72 h. Molecular docking was performed and quercetin had the ideal interaction score values, so it was used with the PACAP protein which presented a slightly moderate interaction with the protein synthesis of Schwann cells, presenting compactness of the compound after 14 ns.

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Colangelo, Maria Teresa, Silvana Belletti, Paolo Govoni, Stefano Guizzardi, and Carlo Galli. "A Biomimetic Polynucleotides–Hyaluronic Acid Hydrogel Promotes Wound Healing in a Primary Gingival Fibroblast Model." Applied Sciences 11, no. 10 (May 12, 2021): 4405. http://dx.doi.org/10.3390/app11104405.

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Polynucleotides (PN) have long been known as an effective supportive therapy for wound healing. The present study investigated whether a hydrogel formulation containing PN and hyaluronic acid (PN + HA) could promote wound healing in an in vitro model of gingival fibroblasts. PN promoted cell growth and viability as assessed by different assays, and PN + HA, though not significantly further increasing cell growth as compared to PN, supported the formation of dense multilayered cell nodules. PN promoted fibroblasts’ clonogenic efficiency and PN + HA further enhanced the formation of more numerous dense colonies. PN + HA appeared to significantly increase the expression of collagen 1a1 and 3a1, while not affecting proteoglycans deposition. Interestingly, when tested in a scratch assay, PN + HA achieved gap closure after 48 h, while cells in the comparison groups had not completely bridged the scratch even after 96 h. Taken together, these results demonstrate that PN + HA is a promising candidate for a supportive therapy to promote soft tissue healing in the oral cavity.

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Browning, Alexander P., Wang Jin, Michael J. Plank, and Matthew J. Simpson. "Identifying density-dependent interactions in collective cell behaviour." Journal of The Royal Society Interface 17, no. 165 (April 2020): 20200143. http://dx.doi.org/10.1098/rsif.2020.0143.

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Scratch assays are routinely used to study collective cell behaviour in vitro . Typical experimental protocols do not vary the initial density of cells, and typical mathematical modelling approaches describe cell motility and proliferation based on assumptions of linear diffusion and logistic growth. Jin et al. (Jin et al . 2016 J. Theor. Biol. 390 , 136–145 ( doi:10.1016/j.jtbi.2015.10.040 )) find that the behaviour of cells in scratch assays is density-dependent, and show that standard modelling approaches cannot simultaneously describe data initiated across a range of initial densities. To address this limitation, we calibrate an individual-based model to scratch assay data across a large range of initial densities. Our model allows proliferation, motility, and a direction bias to depend on interactions between neighbouring cells. By considering a hierarchy of models where we systematically and sequentially remove interactions, we perform model selection analysis to identify the minimum interactions required for the model to simultaneously describe data across all initial densities. The calibrated model is able to match the experimental data across all densities using a single parameter distribution, and captures details about the spatial structure of cells. Our results provide strong evidence to suggest that motility is density-dependent in these experiments. On the other hand, we do not see the effect of crowding on proliferation in these experiments. These results are significant as they are precisely the opposite of the assumptions in standard continuum models, such as the Fisher–Kolmogorov equation and its generalizations.

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Veres-Székely, Apor, Domonkos Pap, Beáta Szebeni, László Őrfi, Csenge Szász, Csenge Pajtók, Eszter Lévai, Attila J. Szabó, and Ádám Vannay. "Transient Agarose Spot (TAS) Assay: A New Method to Investigate Cell Migration." International Journal of Molecular Sciences 23, no. 4 (February 14, 2022): 2119. http://dx.doi.org/10.3390/ijms23042119.

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Fibroblasts play a central role in diseases associated with excessive deposition of extracellular matrix (ECM), including idiopathic pulmonary fibrosis. Investigation of different properties of fibroblasts, such as migration, proliferation, and collagen-rich ECM production is unavoidable both in basic research and in the development of antifibrotic drugs. In the present study we developed a cost-effective, 96-well plate-based method to examine the migration of fibroblasts, as an alternative approach to the gold standard scratch assay, which has numerous limitations. This article presents a detailed description of our transient agarose spot (TAS) assay, with instructions for its routine application. Advantages of combined use of different functional assays for fibroblast activation in drug development are also discussed by examining the effect of nintedanib—an FDA approved drug against IPF—on lung fibroblasts.

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Gao, Hong, Peipei Tang, Kejie Ni, Lun Zhu, Song Chen, Yulong Zheng, and Yufeng Wan. "Inhibition of Kelch-like epichlorohydrin-related protein 1 promotes the progression and drug resistance of lung adenocarcinoma." PeerJ 9 (August 19, 2021): e11908. http://dx.doi.org/10.7717/peerj.11908.

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Background Lung cancer is a common malignant carcinoma of respiratory system with high morbidity and mortality. Kelch-like epichlorohydrin-related protein 1 (Keap1), a member of the BTB-Kelch protein family, has been reported as an important molecule in several cancers. However, its potential role in tumor is still controversial. Here we aim to clarify the effect of Keap1 on the biological characteristics and chemotherapy resistance in lung adenocarcinoma (LUAD). Methods Immunohistochemistry was conducted to compare Keap1 expression in lung adenocarcinoma tissues and matched non-cancerous tissues, and the correlation between Keap1 expression and clinicopathological features was analyzed. Subsequently, the stable A549 and H1299 cell lines with Keap1 knockdown or overexpression were constructed using lentivirus. The roles of Keap1 on the cell proliferation, migration, invasion and drug resistance were investigated by colony formation assay, cell proliferation assay, wound scratch test, transwell invasion assay and drug sensitivity assay, respectively. Results Keap1 was lowly expressed in tumor tissues compared to matched non-cancerous tissues, and its expression was correlated with TNM stage and lymph node metastasis. Early stage (I) tumors without lymph node metastasis had higher levels of Keap1 expression compared with late-stage tumors (II, III) with the presence of lymphatic metastasis. Colony formation assays showed that Keap1 knockdown promoted the proliferation of A549 and H1299 cells, and the cell growth curves further confirmed this feature. In contrast, wound scratch and transwell invasion experiments showed that Keap1 overexpression inhibited cell migration and invasive malignancy. The IC50 for cisplatin and paclitaxel were significantly increased by Keap1 knockdown in A549 and H1299 cell lines. Conclusion Keap1 knockdown promotes tumor cell growth, proliferation, invasion, metastasis and chemotherapy resistance in LUAD. It may be a potential tumor marker to guide the staging and treatment of lung cancer.

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Hopper, Niina, John Wardale, Daniel Howard, Roger Brooks, Neil Rushton, and Frances Henson. "Peripheral Blood Derived Mononuclear Cells Enhance the Migration and Chondrogenic Differentiation of Multipotent Mesenchymal Stromal Cells." Stem Cells International 2015 (2015): 1–9. http://dx.doi.org/10.1155/2015/323454.

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A major challenge in cartilage repair is the lack of chondrogenic cells migrating from healthy tissue into damaged areas and strategies to promote this should be developed. The aim of this study was to evaluate the effect of peripheral blood derived mononuclear cell (PBMC) stimulation on mesenchymal stromal cells (MSCs) derived from the infrapatellar fat pad of human OA knee. Cell migration was measured using an xCELLigence electronic migration chamber system in combination with scratch assays. Gene expression was quantified with stem cell PCR arrays and validated using quantitative real-time PCR (rtPCR). In both migration assays PBMCs increased MSC migration by comparison to control. In scratch assay the wound closure was 55% higher after 3 hours in the PBMC stimulated test group (P=0.002), migration rate was 9 times faster (P=0.008), and total MSC migration was 25 times higher after 24 hours (P=0.014). Analysis of MSCs by PCR array demonstrated that PBMCs induced the upregulation of genes associated with chondrogenic differentiation over 15-fold. In conclusion, PBMCs increase both MSC migration and differentiation suggesting that they are an ideal candidate for inclusion in regenerative medicine therapies aimed at cartilage repair.

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Gersch, Robert P., Jeffrey C. Raum, Catherine Calvert, and Ivona Percec. "Fibroblasts Derived From Human Adipose Stem Cells Produce More Effective Extracellular Matrix and Migrate Faster Compared to Primary Dermal Fibroblasts." Aesthetic Surgery Journal 40, no. 1 (March 15, 2019): 108–17. http://dx.doi.org/10.1093/asj/sjz071.

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Abstract Background The efficacy of adipose-derived stem cells (ASCs) to improve wound healing has been extensively investigated. Unfortunately, no consistent reports have described significant improvements in healing time or outcomes in large-scale clinical trials utilizing human ASCs. Primarily, these studies examined undifferentiated ASCs as opposed to specific cells differentiated from ASCs. Objectives The authors sought to examine the properties of fibroblasts differentiated from human ASCs (dFib cells) compared with those of primary dermal fibroblasts. Methods ASCs were isolated from healthy female patients, differentiated into dFib cells, and compared with intra-patient primary dermal fibroblasts for morphology, extracellular matrix (ECM) marker expression, and cell migration employing qPCR, western blot, and scratch test assays. Results De novo differentiated fibroblasts produce higher levels of the healthy ECM markers Elastin, Fibronectin, and Collagen 1 compared with primary fibroblasts. In contrast, dFib cells have reduced expression of the scar tissue markers αSMA, Collagen 3, and MMP-1. Further, dFib cells close scratch defects more quickly than primary dermal fibroblasts (32 ± 12.85 hours vs 64 ± 13.85 hours, P < 0.01) in a scratch test assay. Conclusions These data suggest that fibroblasts newly differentiated from human ASCs migrate well and produce a robust ECM, the combination of which may contribute to improved wound healing, and thus should be further investigated.

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Lin, Lei, Hong-bin Tu, Liang Wu, Ming Liu, and Ge-ning Jiang. "MicroRNA-21 Regulates Non-Small Cell Lung Cancer Cell Invasion and Chemo-Sensitivity through SMAD7." Cellular Physiology and Biochemistry 38, no. 6 (2016): 2152–62. http://dx.doi.org/10.1159/000445571.

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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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Choi, Sun-Hye, Kyung-Jong Won, Rami Lee, Han-Sung Cho, Sung-Hee Hwang, and Seung-Yeol Nah. "Wound Healing Effect of Gintonin Involves Lysophosphatidic Acid Receptor/Vascular Endothelial Growth Factor Signaling Pathway in Keratinocytes." International Journal of Molecular Sciences 22, no. 18 (September 21, 2021): 10155. http://dx.doi.org/10.3390/ijms221810155.

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Gintonin, a novel compound of ginseng, is a ligand of the lysophosphatidic acid (LPA) receptor. The in vitro and in vivo skin wound healing effects of gintonin remain unknown. Therefore, the objective of this study was to investigate the effects of gintonin on wound healing-linked responses, especially migration and proliferation, in skin keratinocytes HaCaT. In this study, 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide assay, Boyden chamber migration assay, scratch wound healing assay, and Western blot assay were performed. A tail wound mouse model was used for the in vivo test. Gintonin increased proliferation, migration, and scratch closure in HaCaT cells. It also increased the release of vascular endothelial growth factor (VEGF) in HaCaT cells. However, these increases, induced by gintonin, were markedly blocked by treatment with Ki16425, an LPA inhibitor, PD98059, an ERK inhibitor, 1,2-Bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid tetrakis (acetoxymethyl ester), a calcium chelator, and U73122, a PLC inhibitor. The VEGF receptor inhibitor axitinib also attenuated gintonin-enhanced HaCaT cell proliferation. Gintonin increased the phosphorylation of AKT and ERK1/2 in HaCaT cells. In addition, gintonin improved tail wound healing in mice. These results indicate that gintonin may promote wound healing through LPA receptor activation and/or VEGF release-mediated downstream signaling pathways. Thus, gintonin could be a beneficial substance to facilitate skin wound healing.

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Lestari, Beni, Laeli Muntafiah, Ziana Walidah, and Riris Istighfari Jenie. "A Comparison of Antimetastatic Activity between Nerium indicum and Cinnamomum burmannii on 4T1 Cells." Indonesian Journal of Cancer Chemoprevention 8, no. 2 (June 30, 2017): 85. http://dx.doi.org/10.14499/indonesianjcanchemoprev8iss2pp85-93.

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Metastatic process becomes a major problem in advanced cancer cases. Natural compounds found in several plants in Indonesia have a potency to be developed as chemoterapeutic agent which are targeted to metastatic process. Jure leaves (Nerium indicum) which contain oleandrin and cinnamaldehyde in cinnamon bark (Cinnamomum burmannii) reported to have cytotoxic activity on several cancer cells, but their activities on metastasic process have never been explored. This research aims to reveal and to compare their anti-metastatic effect toward 4T1 breast cancer cells. The cytotoxicity of jure leaves extract (JLE) and cinnamon essential oil (CEO) was obtained by MTT assay. Metastatic process mainly on cell migration was examined by scratch wound healing assay while MMP-9 expression that described the invassion process was observed by gelatin zymography assay. Molecular interaction between their active compounds and MMP-9 receptor was predicted by molecular docking. The result showed that treatment with JLE and CEO inhibited the growth of 4T1 cells with IC50 value of 125 µg/mL and 2.5 µg/mL, respectively. In addition, JLE performed inhibitory effect of cell migration better than CEO. Meanwhile, both JLE and CEO decreased MMP-9 protein expression. Thus, JLE and CEO have potentials to be developed as an anti-metastatic agent and JLE could be more effective.Key words: Nerium indicum, Cinnamomum burmannii, anti-metastasis, scratch assay, gelatin zimography

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Johnston, Stuart T., Matthew J. Simpson, D. L. Sean McElwain, Benjamin J. Binder, and Joshua V. Ross. "Interpreting scratch assays using pair density dynamics and approximate Bayesian computation." Open Biology 4, no. 9 (September 2014): 140097. http://dx.doi.org/10.1098/rsob.140097.

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Quantifying the impact of biochemical compounds on collective cell spreading is an essential element of drug design, with various applications including developing treatments for chronic wounds and cancer. Scratch assays are a technically simple and inexpensive method used to study collective cell spreading; however, most previous interpretations of scratch assays are qualitative and do not provide estimates of the cell diffusivity, D , or the cell proliferation rate, λ . Estimating D and λ is important for investigating the efficacy of a potential treatment and provides insight into the mechanism through which the potential treatment acts. While a few methods for estimating D and λ have been proposed, these previous methods lead to point estimates of D and λ , and provide no insight into the uncertainty in these estimates. Here, we compare various types of information that can be extracted from images of a scratch assay, and quantify D and λ using discrete computational simulations and approximate Bayesian computation. We show that it is possible to robustly recover estimates of D and λ from synthetic data, as well as a new set of experimental data. For the first time, our approach also provides a method to estimate the uncertainty in our estimates of D and λ . We anticipate that our approach can be generalized to deal with more realistic experimental scenarios in which we are interested in estimating D and λ , as well as additional relevant parameters such as the strength of cell-to-cell adhesion or the strength of cell-to-substrate adhesion.

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Gnerucci, Alessio, Paola Faraoni, Elettra Sereni, and Francesco Ranaldi. "Scratch assay microscopy: A reaction–diffusion equation approach for common instruments and data." Mathematical Biosciences 330 (December 2020): 108482. http://dx.doi.org/10.1016/j.mbs.2020.108482.

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Goetsch, K. P., and C. U. Niesler. "Optimization of the scratch assay for in vitro skeletal muscle wound healing analysis." Analytical Biochemistry 411, no. 1 (April 2011): 158–60. http://dx.doi.org/10.1016/j.ab.2010.12.012.

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Orfanoudaki, Maria, Anja Hartmann, Mostafa Alilou, Thomas Gelbrich, Patricia Planchenault, Séverine Derbré, Andreas Schinkovitz, Pascal Richomme, Andreas Hensel, and Markus Ganzera. "Absolute Configuration of Mycosporine-Like Amino Acids, Their Wound Healing Properties and In Vitro Anti-Aging Effects." Marine Drugs 18, no. 1 (December 31, 2019): 35. http://dx.doi.org/10.3390/md18010035.

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Mycosporine-like amino acids (MAAs) are water-soluble metabolites, reported to exhibit strong UV-absorbing properties. They have been found in a wide range of marine organisms, especially those that are exposed to extreme levels of sunlight, to protect them against solar radiation. In the present study, the absolute configuration of 14 mycosporine-like-amino acids was determined by combining the results of electronic circular dichroism (ECD) experiments and that of advanced Marfey’s method using LC-MS. The crystal structure of a shinorine hydrate was determined from single crystal X-ray diffraction data and its absolute configuration was established from anomalous-dispersion effects. Furthermore, the anti-aging and wound-healing properties of these metabolites were evaluated in three different assays namely the inhibition of collagenase, inhibition of advanced glycation end products (AGEs) and wound healing assay (scratch assay).

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Bahar, Entaz, and Hyonok Yoon. "Modeling and Predicting the Cell Migration Properties from Scratch Wound Healing Assay on Cisplatin-Resistant Ovarian Cancer Cell Lines Using Artificial Neural Network." Healthcare 9, no. 7 (July 19, 2021): 911. http://dx.doi.org/10.3390/healthcare9070911.

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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.

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Quintão, Glauber Cisneiros, Giani Maria Cavalcante, and Luiz Andrade Lins. "Investigation of wound healing in vitro of specie Brachymenium exile (Dozy & Molk.) Bosch & Sande Lac." Research, Society and Development 11, no. 4 (March 11, 2022): e6311427057. http://dx.doi.org/10.33448/rsd-v11i4.27057.

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The Brachymenium exile is the species of Bryaceae family, the family of bryophytes best represented in cerrado biome. The potential for biotechnological and biopharmaceutical applications of bryophytes have been investigated sharply. The aimed to investigate the cicatrizing potential of ethanolic extract of B. exile by scratch assay. The in vitro study was using the spreading and migration capabilities of fibroblast cell line using the extract of B. exile. the extract of B. exile allowed the cellular migration in 76% more that the control and not citotoxic assay in fibroblast cell line (3T3-L1). The extract of B. exile showed activity potential wound healing in vitro assay.

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Laheij, Alexa M. G. A., Johannes J. de Soet, Enno C. I. Veerman, Jan G. M. Bolscher, and Cor van Loveren. "The Influence of Oral Bacteria on Epithelial Cell MigrationIn Vitro." Mediators of Inflammation 2013 (2013): 1–6. http://dx.doi.org/10.1155/2013/154532.

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Oral ulcerations often arise as a side effect from chemo- and radiation therapy. In a previous clinical study,Porphyromonas gingivaliswas identified as a positive predictor for oral ulcerations after hematopoetic stem cell transplantation, possibly incriminatingP. gingivalisin delayed healing of the ulcerations. Therefore, it was tested whetherP. gingivalisand its secreted products could inhibit the migration of oral epithelial cells in anin vitroscratch assay. To compare, the oral bacteriaPrevotella nigrescens,Prevotella intermedia,Tannerella forsythia, andStreptococcus mitiswere included. A standardized scratch was made in a confluent layer of human oral epithelial cells. The epithelial cells were challenged with bacterial cells and with medium containing secretions of these bacteria. Closure of the scratch was measured after 17 h using a phase contrast microscope.P. gingivalis,P. nigrescens, and secretions ofP. gingivalisstrongly inhibited cell migration. A challenge with 1000 heat-killed bacteria versus 1 epithelial cell resulted in a relative closure of the scratch of 25% forP. gingivalisand 20% forP. nigrescens. Weaker inhibitory effects were found for the other bacteria. The results confirmed our hypothesis that the oral bacteria may be involved in delayed wound healing.

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Journal articles on the topic 'Scratch Assay'

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