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1
Alishahedani, Mohammadali E., Manoj Yadav, Katelyn J. McCann, et al. "Therapeutic candidates for keloid scars identified by qualitative review of scratch assay research for wound healing." PLOS ONE 16, no. 6 (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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Trivedi, Mahendra Kumar, Alice Branton, Dahryn Trivedi, Gopal Nayak, Mayank Gangwar, and Snehasis Jana. "Wound Healing Potential of Biofield Energy Treatment in HaCaT Cell Line Using Scratch Assay." Global Research Journal of Public Health and Epidemiology 5, no. 5 (2018): 136–42. https://doi.org/10.5281/zenodo.2560883.
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Wounds may be acute, subacute, or chronic and are the common clinical life entities. It is a complex biochemical process to restore the cell structures that depends upon cell proliferation and migration, mainly by keratinocytes and fibroblast cells. The aim of the present study was to evaluate the effect of the Biofield Energy Treatment (Consciousness Energy Healing Treatment-The Trivedi Effect®) in the HaCaT cell line (Human keratinocytes) and DMEM (Dulbecco’s Modified Eagle Medium) for wound healing action using scratch assay (48 hours) against positive control i.e., recombinant Human Epidermal Growth Factor (Hu-EGF, 30 ng/mL). In vitro scratch assay monitored proliferation and migration of keratinocytes in the scratched wounded area using WimScratch Image analysis software. The results showed that the Biofield Energy Treated DMEM group have significantly higher percentage of keratinocytes migration i.e., 35.4%, while no migration was monitored in the Biofield Energy Treated HaCaT cells as compared with the baseline control group. Additionally, the percentage of scratch area was significantly decreased by 7.3% in the Biofield Energy Treated DMEM, while an increased scratched area was reported by 14% in the Biofield Energy Treated cells. Hence, the results using scratch assay showed a promising scientific approach to differentiate the Biofield Energy Treatment on cells or media for their wound healing potential. The data concluded that The Trivedi Effect® has the significant capability in wound healing activity in DMEM as compared with the HaCaT cell line with respect to the cell migration and reduced wound scratched area. Consciousness Energy Healing Treatment (The Trivedi Effect®) can be used as a complementary and alternative approach to improve the cellular migration and proliferation and can be used in psoriasis, seborrheic dermatitis, skin cancer, rashes from bacterial or fungal infections as anti-wrinkling, skin-whitening, anti-ageing, and rejuvenating agent. <strong>Source</strong>: https://www.trivedieffect.com/science/wound-healing-potential-of-biofield-energy-treatment-in-hacat-cell-line-using-scratch-assay http://springjournals.net/grjphe/springjournals.netgrjphearticlesindex=4trivedietal
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Trivedi, Mahendra Kumar, Alice Branton, Dahryn Trivedi, Gopal Nayak, Mayank Gangwar, and Snehasis Jana. "Wound Healing Potential of Biofield Energy Treatment in HaCaT Cell Line Using Scratch Assay." Global Research Journal of Public Health and Epidemiology 5, no. 5 (2019): 136–42. https://doi.org/10.5281/zenodo.2566679.
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Wounds may be acute, subacute, or chronic and are the common clinical life entities. It is a complex biochemical process to restore the cell structures that depends upon cell proliferation and migration, mainly by keratinocytes and fibroblast cells. The aim of the present study was to evaluate the effect of the Biofield Energy Treatment (Consciousness Energy Healing Treatment-The Trivedi Effect®) in the HaCaT cell line (Human keratinocytes) and DMEM (Dulbecco’s Modified Eagle Medium) for wound healing action using scratch assay (48 hours) against positive control i.e., recombinant Human Epidermal Growth Factor (Hu-EGF, 30 ng/mL). In vitro scratch assay monitored proliferation and migration of keratinocytes in the scratched wounded area using WimScratch Image analysis software. The results showed that the Biofield Energy Treated DMEM group have significantly higher percentage of keratinocytes migration i.e., 35.4%, while no migration was monitored in the Biofield Energy Treated HaCaT cells as compared with the baseline control group. Additionally, the percentage of scratch area was significantly decreased by 7.3% in the Biofield Energy Treated DMEM, while an increased scratched area was reported by 14% in the Biofield Energy Treated cells. Hence, the results using scratch assay showed a promising scientific approach to differentiate the Biofield Energy Treatment on cells or media for their wound healing potential. The data concluded that The Trivedi Effect® has the significant capability in wound healing activity in DMEM as compared with the HaCaT cell line with respect to the cell migration and reduced wound scratched area. Consciousness Energy Healing Treatment (The Trivedi Effect®) can be used as a complementary and alternative approach to improve the cellular migration and proliferation and can be used in psoriasis, seborrheic dermatitis, skin cancer, rashes from bacterial or fungal infections as anti-wrinkling, skin-whitening, anti-ageing, and rejuvenating agent. https://www.trivedieffect.com/science/wound-healing-potential-of-biofield-energy-treatment-in-hacat-cell-line-using-scratch-assay http://springjournals.net/grjphe/springjournals.netgrjphearticlesindex=4trivedietal
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Trivedi, Mahendra Kumar, Alice Branton, Dahryn Trivedi, Gopal Nayak, Mayank Gangwar, and Snehasis Jana. "Wound Healing Activity of Consciousness Energy Healing Treatmenton HFF-1 Cells and DMEM Using Scratch Assay." Investigative Dermatology and Venereology Research 4, no. 1 (2018): 50–54. https://doi.org/10.5281/zenodo.3374350.
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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 activity via 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. <strong>Source:</strong> https://www.trivedieffect.com/science/wound-healing-activity-of-consciousness-energy-healing-treatmenton-hff-1-cells-and-dmem-using-scratch-assay/ https://www.ommegaonline.org/article-details/Wound-Healing-Activity-of-Consciousness-Energy-Healing-Treatmenton-HFF-1-Cells-and-DMEM-Using-Scratch-Assay-/2036
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Amalia, Latifa, Retno Murwanti, Triana Hertiani, and Kurnia Rahayu Purnomo. "Evaluation of the Potential In Vitro effects of Plantago major L. on Wound Healing in Human Umbilical Vein Endothelial Cells (HUVEC)." Indonesian Journal of Cancer Chemoprevention 15, no. 2 (2025): 87. https://doi.org/10.14499/indonesianjcanchemoprev15iss2pp87-95.
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The treatment of skin wounds remains a major concern in the field of medicine, particularly in the case of chronic wounds resulting from various disorders such as diabetes. The utilization of herbs or herbal preparations for the purpose of healing skin wounds presents a therapeutic challenge within the realm of traditional medicine. Plantago major L. is known to have bioactive compounds that have wound healing activity such as aucubin. This study aimed to determine the in vitro wound healing potential of Plantago major L. extract (PLE). The study involved several assays, including phytochemical examination of PLE using TLC, cell viability testing using MTT assay, and wound healing testing using scratch assay on human umbilical vein endothelial cells (HUVEC). The results confirmed the presence of aucubin as one of the compounds in PLE. It was observed that PLE with 125 μg/mL exhibited the highest wound closure percentage at 90.66%. This study shows that PLE possesses wound healing capabilities.Keywords: Plantago major L., PLE, cytotoxic assay, wound scratch assay, HUVEC.
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Balko, Stefan, Evan Kerr, Edward Buchel, Sarvesh Logsetty, and Afshin Raouf. "A Robust and Standardized Approach to Quantify Wound Closure Using the Scratch Assay." Methods and Protocols 6, no. 5 (2023): 87. http://dx.doi.org/10.3390/mps6050087.
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The scratch assay is an in vitro assay that allows for high-throughput quantification of wound closure by keratinocytes and fibroblasts with relative ease. However, this assay is amenable to experimental variables, which can result in false-positive and false-negative data, making the interpretation of such data difficult. Also, data variability decreases the sensitivity of the scratch assay. Here, we identify important sources of data variation in the scratch assay and provide rational mitigation strategies that enable robust and highly reproducible quantification of scratch width and area, and ultimately the scratch closure rates. By eliminating these sources of variability, the sensitivity of the scratch assay is enhanced, thereby allowing for identification of dependent variables with wide-ranging impacts on wound closure in a robust and standardized manner.
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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 (2018): 50–54. http://dx.doi.org/10.15436/2381-0858.18.2036.
Full textAbstract:
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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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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Azis, Huwaida Abdul, Muhammad Taher, Deny Susanti, and Zainul Amiruddin Zakaria. "IN VITRO WOUND HEALING ACTIVITY OF SAPIUM INDICUM WILLD LEAF EXTRACTS." African Journal of Traditional, Complementary and Alternative Medicines 12, no. 2 (2015): 126–32. http://dx.doi.org/10.21010/ajtcam.v12i2.19.
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Background: Sapium indicum Willd (Euphorbiaceae) is traditionally used to accelerate wound healing process in Malay community.
 Objective: To evaluate wound healing potential of aqueous (AESI) and ethanolic extract of S. indicum leaves (EESI) using cell migration model. 
 Methods: AESI and EESI were prepared using maceration techniques. The extracts were subjected to phytochemical screening, cytotoxicity (MTT assay) and scratch assay.
 Results: AESI contained saponins and tannins while EESI contained triterpenes, tannins and proteins. Based on MTT assay, AESI and EESI exhibited IC50 = 16.3 and 70.9 µg/mL, respectively. In the scratch assay, EESI caused significant (P< 0.05) concentration-dependent migration on 3T3 L1 cells whereas AESI exerted concentration-independent effect. 
 Conclusion: The leaves of S. indicum exhibited wound healing potential and contained phytochemicals that may contribute to the activity. These findings would support potential use of S. indicum as wound healing plant.
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Radstake, Wilhelmina E., Kiran Gautam, Cynthia Van Rompay, et al. "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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Shekatkar, Madhura, Supriya Kheur, Shantanu Deshpande, et al. "Enhancing angiogenesis through secretomes: Insights from scratch wound assay." Journal of Oral Biology and Craniofacial Research 15, no. 4 (2025): 789–98. https://doi.org/10.1016/j.jobcr.2025.05.008.
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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 (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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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 (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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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 (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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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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Gough, Wendy, Keren I. Hulkower, Renee Lynch, et al. "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 (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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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 (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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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 (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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Dogra, Shivani, Bhupendra Koul, Joginder Singh, Meerambika Mishra, and Dhananjay Yadav. "Phytochemical Analysis, Antimicrobial Screening and In Vitro Pharmacological Activity of Artemisia vestita Leaf Extract." Molecules 29, no. 8 (2024): 1829. http://dx.doi.org/10.3390/molecules29081829.
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Artemisia vestita Wall. Ex Besser is a folklore medicinal plant that belongs to Asteraceae family and a treasure trove of drugs. The aim of this research study was to investigate the phytoconstituents, antimicrobial activity, antioxidant, anti-inflammatory, cytotoxicity and wound healing potential of A. vestita leaf extract (ALE). Phytochemical analysis of the ALE was carried out by Soxhlet extraction and GCMS (gas chromatography–mass spectrometry) analysis. Antimicrobial activity was performed by the agar well diffusion method against selected bacterial and fungal strains. Free radical scavenging potential was evaluated by DPPH (2,2-Diphenyl-1-picrylhydrazyl), ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) and FRAP (Ferric reducing antioxidant power) assays. Anti-inflammatory activity was performed by enzyme inhibition assay–COXII. The cytotoxicity of ALE on HaCaT cells was studied via MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay. An in vitro scratch assay was performed for the evaluation of the wound healing property of ALE. It showed satisfactory antimicrobial activity against Staphylococcus aureus (14.2 ± 0.28 mm), Escherichia coli (17.6 ± 0.52 mm), Bacillus subtilis (13.1 ± 0.37 mm), Streptococcus pyogenes (17.3 ± 0.64 mm), Proteus mirabilis (9.4 ± 0.56 mm), Aspergillus niger (12.7 ± 0.53 mm), Aspergilus flavus (15.3 ± 0.25 mm) and Candida albicans (17.6 ± 0.11 mm). In ALE, 36 phytochemicals were detected by GCMS analysis, but 22 were dominant. Moreover, the ALE was effective in scavenging free radicals with different assays and exhibited reasonable anti-inflammatory activity. The MTT assay revealed that ALE had a cytotoxic effect on the HaCaT cells. The scratch assay showed 94.6% wound closure (after 24 h incubation) compared to the positive control Cipladine, which is remarkable wound healing activity. This is the first report on the wound healing property of A. vestita, which can serve as a potential agent for wound healing and extends knowledge on its therapeutic potential.
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Abbas, ManalAhmad, ManalMohammad Abbas, Naseer Al-Rawi та Iqbal Al-Khateeb. "Naringenin potentiated β-sitosterol healing effect on the scratch wound assay". Research in Pharmaceutical Sciences 14, № 6 (2019): 566. http://dx.doi.org/10.4103/1735-5362.272565.
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Rizqi, Januar, Akbar Satria Fitriawan, and Ririn Wahyu Widayati. "THE EFFECTIVENESS OF INDONESIAN HONEY STIMULATE FIBROBLAST CELL VIABILITY AND MIGRATION THAT COULD POTENTIAL PROMOTE WOUND HEALING." Jurnal Keperawatan Respati Yogyakarta 9, no. 1 (2022): 51. http://dx.doi.org/10.35842/jkry.v9i1.652.
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Wound healing is a complex event involving both cellular and molecular activities. Fibroblasts play an important role and are keys to wound healing through cell proliferation and migration. Honey has anti-microbial, anti-oxidant, anti-inflammatory properties, which are used for various benefits such as wound healing. This study aims to explore the effect of honey on the viability and migration ability of fibroblast cells. The method used is the viability test using the MTT Assay calculated by the formula for the percentage of cell viability. Migration test using In Vitro Wound Scratch Assay. The results of the migration test images were analyzed using ImageJ. Giving honey doses of 0.5% and 0.1% increased cell viability and migration after 24 hours of intervention. Decreased cell viability after 48 hours of treatment, but there was a difference in the meaning of honey 1%, 0.5%, and 0.1% compared to control. Honey doses of 1%, 0.5%, and 0.1% increased fibroblast cell migration compared to control. The lowest honey increases the viability and migration of fibroblasts so that the possibility of wound healing.
 Keywords: Honey; Fibroblast Migration; Wound Scratch Assay; Wound Healing
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Zanchetta, Flávia Cristina, Pieter De Wever, Joseane Morari, et al. "In Vitro and In Vivo Evaluation of Chitosan/HPMC/Insulin Hydrogel for Wound Healing Applications." Bioengineering 11, no. 2 (2024): 168. http://dx.doi.org/10.3390/bioengineering11020168.
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Treatment of chronic wounds is challenging, and the development of different formulations based on insulin has shown efficacy due to their ability to regulate oxidative stress and inflammatory reactions. The formulation of insulin with polysaccharides in biohybrid hydrogel systems has the advantage of synergistically combining the bioactivity of the protein with the biocompatibility and hydrogel properties of polysaccharides. In this study, a hydrogel formulation containing insulin, chitosan, and hydroxypropyl methyl cellulose (Chi/HPMC/Ins) was prepared and characterized by FTIR, thermogravimetric, and gel point analyses. The in vitro cell viability and cell migration potential of the Chi/HPMC/Ins hydrogel were evaluated in human keratinocyte cells (HaCat) by MTT and wound scratch assay. The hydrogel was applied to excisional full-thickness wounds in diabetic mice for twenty days for in vivo studies. Cell viability studies indicated no cytotoxicity of the Chi/HPMC/Ins hydrogel. Moreover, the Chi/HPMC/Ins hydrogel promoted faster gap closure in the scratch assay. In vivo, the wounds treated with the Chi/HPMC/Ins hydrogel resulted in faster wound closure, formation of a more organized granulation tissue, and hair follicle regeneration. These results suggest that Chi/HPMC/Ins hydrogels might promote wound healing in vitro and in vivo and could be a new potential dressing for wound healing.
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Alsareii, Saeed Ali, Nasser A. N. Alzerwi, Mansour Yousef AlAsmari, Abdulrahman Manaa Alamri, Mater H. Mahnashi, and Ibrahim Ahmed Shaikh. "Topical Application of Premna integrifolia Linn on Skin Wound Injury in Rats Accelerates the Wound Healing Process: Evidence from In Vitro and In Vivo Experimental Models." Evidence-Based Complementary and Alternative Medicine 2022 (April 13, 2022): 1–14. http://dx.doi.org/10.1155/2022/6449550.
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Background. When the skin and tissues within the body are injured, the healing process begins. Medicinal herbs have been used to cure wounds since time immemorial. The antimicrobial and antioxidant activity possessed by P. integrifolia may accelerate wound healing. Objectives. To assess the wound healing activity of Premna integrifolia extract (PIE) by employing in-vivo experimental animal models and an in-vitro migration scratch assay. Furthermore, to assess its cytotoxicity using the MTT assay. Methods. Wistar albino rats were used for the in vivo wound healing models. The animals were divided into four groups at random: Group I was untreated. Group II was vehicle control (ointment base). Group III was PIE ointment (5% W/W). Group IV was standard (povidone-iodine ointment) (5% W/W). The ointments were applied directly to the wounds as described above until they healed completely. The wound contraction percentage and tensile strength were calculated. The MTT test was used to determine the viability of the test extract against the fibroblast cells. The scratch assay was used in vitro to determine the wound healing potential of the test drug. P ≤ 0.05 values were considered statistically significant. Results. Premna integrifolia extract did not possess any noticeable cytotoxicity to the cell line and showed an IC50 of 185.98 μg/ml. The wound contraction potential of PIE ointment-treated animals was considerably greater ( P ≤ 0.001 ) on days 4, 8, 12, 16, and 20 when compared to the control group. The percentage of wound contraction on day 20 was 99.92% in PIE-treated animals compared to 83.23% in untreated animals. Compared to the untreated group, the duration of full epithelization was significantly ( P ≤ 0.01 ) shorter in the test group. When compared to the incision control group, the animals treated with PIE ointment had significantly higher ( P ≤ 0.001 ) tensile strength. In addition, animals given the test drug had a significant ( P ≤ 0.001 ) increase in total protein and hydroxyproline. In the in vitro scratch assay, test drug-treated cells demonstrated greater cell migration. Histology images confirmed that the test drug-treated group had epithelial tissue proliferation and keratinization. Conclusion. The current study found that Premna integrifolia improved wound healing activity both in vitro and in vivo. These findings indicate that Premna integrifolia extract has wound-healing potential and could be a viable source of nutraceuticals with wound-healing properties.
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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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Bakari, G. G., S. A. Mshamu, M. H. Ally, R. A. Max, and H. Bai. "In-vitro Wound Healing Properties of Commiphora swynnertonii Resinous Extracts." Tanzania Veterinary Journal 38 (September 4, 2021): 32–37. http://dx.doi.org/10.4314/tvj.v38i1.6s.
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Wound healing is a complex multicellular process involving many cell types which include; inflammatory cells, endothelial cells, fibroblasts and keratinocytes. The process involves an orderly sequence of events with four overlapping phases namely; haemostasis, inflammatory, proliferation and remodeling phases. The process can be facilitated by the use of wound healing agents including herbal remedies from plants. In this study the main objective was to evaluate the in vitro wound healing activity of the resin obtained from Commiphora swynnertonii (C.swynnertonii). First the NIH -3T3 cells viability were evaluated using (3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl Tetrazolium Bromide (MTT) assay. Then the wound scratch assay model was used to evaluate cellular proliferation, closure of the wound and release of matrix metalloproteinase enzymes. Results indicate differences in mean cell viability between different concentrations within 24 hours of incubation. The highest viability was recorded at the concentration of 1% (v/v). The in-vitro wound scratch assay showed positive NIH - 3T3 cells proliferation on the wound area and cells migration when compared with control group (without treatment) at 0 and 24 hours. In addition, C. swynnertonii was able to stimulate secretion of MMP-2 release from NIH - 3T3 cells. MMP-2 is an important enzyme for extracellular matrix remodeling during wound healing suggesting that C. swynnertonii promotes wound healing by stimulating cell proliferation and production of MMP-2 in a mechanism that is currently not known.
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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 (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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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 (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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Cheng, Yu, Zhang Hu, Yuntao Zhao, et al. "Sponges of Carboxymethyl Chitosan Grafted with Collagen Peptides for Wound Healing." International Journal of Molecular Sciences 20, no. 16 (2019): 3890. http://dx.doi.org/10.3390/ijms20163890.
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Burns are physically debilitating and potentially fatal injuries. Two marine biomaterials, carboxymethyl chitosan (CMC) and collagen peptides (COP), have emerged as promising burn dressings. In this paper, sponges of carboxymethyl chitosan grafted with collagen peptide (CMC–COP) were prepared by covalent coupling and freeze drying. Scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) spectroscopy were then used to characterize the prepared sponges. To evaluate the wound healing activity of the CMC–COP sponges, in vitro tests including cell viability scratch wound healing and scald wound healing experiments were performed in rabbits. Appearance studies revealed the porous nature of sponges and FTIR spectroscopy demonstrated the successful incorporation of COP into CMC. The in vitro scratch assay showed that treatment with CMC–COP sponges (at 100 μg/mL) had significant effects on scratch closure. For burn wounds treated with CMC–COP, regeneration of the epidermis and collagen fiber deposition was observed on day 7, with complete healing of the epidermis and wound on days 14 and 21, respectively. Based on the pathological examination by hematoxylin and eosinstaining, the CMC–COP group demonstrated pronounced wound healing efficiencies. These results confirmed that the CMC–COP treatment enhanced cell migration and promoted skin regeneration, thereby highlighting the potential application of these sponges in burn care.
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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 (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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Vovk, I. "PRODUCTION AND in vitro EVALUATION OF RECOMBINANT HUMAN RHHB-EGF FOR WOUND HEALING AND TARGETED THERAPY." Biotechnologia Acta 18, no. 1 (2025): 55–66. https://doi.org/10.15407/biotech18.01.055.
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Aim. The goal of the study was to evaluate the biological activity of recombinant human heparinbinding EGF-like growth factor (rhHB-EGF) on mouse fibroblasts in vitro as a potential agent for promoting wound healing and tissue regeneration. Methods. The study employed a scratch assay to evaluate the migration of mouse fibroblasts (L929 and NIH-3T3), the MTT test to assess cell proliferation, MALDI-TOF mass spectrometry for protein identification, and flow cytometry to determine cell viability. Results. In the concentration range of 500-1000 ng/ml rhHB-EGF, no cytotoxic effect was recorded, but an increase in proliferation and/or metabolic activity, as well as migration of fibroblasts, was detected, with a maximum effect at 500 ng/ml rhHB-EGF in the cell incubation medium. A 30% overgrowth of the wound surface of fibroblasts was demonstrated in the scratch assay test under the influence of rhHB-EGF compared to the corresponding control. Conclusions. rhHB-EGF at a concentration of 500 ng/ml can be used in preparations to stimulate wound healing and tissue regeneration due to its ability to stimulate proliferation/metabolic activity and migration of fibroblasts, as well as the lack of cytotoxicity. Further, in vivo studies are needed for a comprehensive evaluation of this possibility.
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Salac, Edcyl Lee O., Michael Russelle Alvarez, Rnie Shayne Gaurana, 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 (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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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 (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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PN, Sruthy, and Saravanan Govindaraj. "Optimization In-vitro Evaluation and Scratch Wound Healing Assay of Hexacosane Topical Gel for Wound Healing." INTERNATIONAL JOURNAL OF DRUG DELIVERY TECHNOLOGY 14, no. 03 (2024): 1361–69. http://dx.doi.org/10.25258/ijddt.14.3.16.
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The compounds hexocosane, a sterol hydrocarbon identified in the dichloromethane extract and isolated from marine sponges of the genus Agelas. Hexocosane possesses anti-inflammatory and antioxidant activities that reduce inflammation and oxidative stress, they promote cell proliferation and angiogenesis, facilitating tissue regeneration and repair. This multifaceted approach makes this secondary metabolite a promising candidate for developing advanced wound care therapies that effectively address both infection control and wound healing. On the basis of viscosity, in-vitro release, and skin retention, optimal gel formulations were developed with hexocosane (1%) to achieve the best results. A response surface methodology Box-Behnken design was applied on three factors and three levels [carbopol 940 (1, 1.25 and 1.5%), hydroxypropyl methylcellulose (HPMC) (1,1.5 and 1.2%), and propylene glycol (0–1–1.5% and 1–2%, respectively] using three factors and three levels. In order to assess the spreadability and viscosity, a glass slide and a Brookfield viscometer were used. An assay was conducted to determine how topical gel affects wound healing. The optimization study of the gel formulation, involving variations in adhesive polymer (Carbopol 940), release retarding polymer (HPMC K4M), and penetration enhancer (Surfactant), has identified three noteworthy formulations (F4, F8 & F14). Each formulation is characterized by specific attributes such as viscosity, in-vitro drug release, and skin retention. All samples were found to elicit significant wound healing efficacy as evidenced from the representative photomicrographs. The maximum efficacy was elicited by formulation (F4) with 100 mg drug at the time duration of 36 hours.
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Dhillon, Prabhpreet K., Xinyin Li, Jurgen T. Sanes, Oluwafemi S. Akintola, and Bingyun Sun. "Method comparison for analyzing wound healing rates." Biochemistry and Cell Biology 95, no. 3 (2017): 450–54. http://dx.doi.org/10.1139/bcb-2016-0163.
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Wound healing scratch assay is a frequently used method to characterize cell migration, which is an important biological process in the course of development, tissue repair, and immune response for example. The measurement of wound healing rate, however, varies among different studies. Here we summarized these measurements into three types: (I) direct rate average; (II) regression rate average; and (III) average distance regression rate. Using Chinese hamster ovary (CHO) cells as a model, we compared the three types of analyses on quantifying the wound closing rate, and discovered that type I & III measurements are more resistant to outliers, and type II analysis is more sensitive to outliers. We hope this study can help researchers to better use this simple yet effective assay.
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Shabestani Monfared, Ghazal, Peter Ertl, and Mario Rothbauer. "Microfluidic and Lab-on-a-Chip Systems for Cutaneous Wound Healing Studies." Pharmaceutics 13, no. 6 (2021): 793. http://dx.doi.org/10.3390/pharmaceutics13060793.
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Cutaneous wound healing is a complex, multi-stage process involving direct and indirect cell communication events with the aim of efficiently restoring the barrier function of the skin. One key aspect in cutaneous wound healing is associated with cell movement and migration into the physically, chemically, and biologically injured area, resulting in wound closure. Understanding the conditions under which cell migration is impaired and elucidating the cellular and molecular mechanisms that improve healing dynamics are therefore crucial in devising novel therapeutic strategies to elevate patient suffering, reduce scaring, and eliminate chronic wounds. Following the global trend towards the automation, miniaturization, and integration of cell-based assays into microphysiological systems, conventional wound healing assays such as the scratch assay and cell exclusion assay have recently been translated and improved using microfluidics and lab-on-a-chip technologies. These miniaturized cell analysis systems allow for precise spatial and temporal control over a range of dynamic microenvironmental factors including shear stress, biochemical and oxygen gradients to create more reliable in vitro models that resemble the in vivo microenvironment of a wound more closely on a molecular, cellular, and tissue level. The current review provides (a) an overview on the main molecular and cellular processes that take place during wound healing, (b) a brief introduction into conventional in vitro wound healing assays, and (c) a perspective on future cutaneous and vascular wound healing research using microfluidic technology.
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Parkar, Hafiza, Olayinka Aiyegoro, Paul Steenkamp, and Vanessa Steenkamp. "Extracts of Terminalia sericea Enhance Cell Migratory Activity of Endothelial Hybrid and Fibroblast Cells In Vitro." Planta Medica 83, no. 18 (2017): 1397–404. http://dx.doi.org/10.1055/s-0043-113324.
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Abstract Terminalia sericea is a plant that has been used amongst others medicinally to treat wounds. The aim of this study was to assess the in vitro wound healing ability of T. sericea. Hot water, methanol, ethyl acetate, and hexane extracts were prepared. Thin layer chromatography (TLC) and ultra-performance liquid chromatography time of flight mass spectrometry (UPLC-TOF-MS) were used to determine the phytochemical classes and genus specific compounds present in the plant. Cytotoxicity was assessed in the SC-1 fibroblast and EA.hy926 endothelial hybrid cell lines using the sulforhodamine B assay. The effect of the extracts on cellular migration in both cell lines was assessed using the scratch assay. The major phytochemical classes detected in the extracts using TLC were alkaloids, coumarins, flavonoids, glycosides, phenolics, saponins, sterols, and terpenoids. The genus-specific compounds punicalagin, sericoside, anolignan B, and arjunic acid were identified in the extracts by means of UPLC-QTOF-MS. Cytotoxicity was not observed after 24 h of exposure and a generally low cytotoxic trend was noted after 72 h. A significant (p < 0.05) enhancement of cell migration in both cell lines was noted in the scratch assay. The wound healing ability of T. sericea is mainly attributed to the migratory and proliferative activity of the extracts responsible for the acceleration of wound closure. Isolation and individualized testing of the active compounds is warranted.
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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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Gorkic, Metka, David Messenger, Linda Wainwright, John Connelly, and Cleo L. Bishop. "P33 Investigating the utility and exploring biological targets of micro-wound healing compounds in 2D and 3D skin models." British Journal of Dermatology 189, no. 1 (2023): e27-e27. http://dx.doi.org/10.1093/bjd/ljad174.054.
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Abstract An injury to the epidermis can result in a micro-wound, characterized by its localization above the dermal–epidermal junction and an absence of vessel damage. Re-epithelialization is driven by keratinocytes, which organize into overlapping migrating and proliferating zones to ensure faithful micro-wound closure. Although still relatively unexplored, keratinocyte-mediated cytokine secretion is thought to be a feature of the micro-wound re-epithelialization response. A failure to re-epithelize, also characteristic of chronic wounds, can lead to reoccurring infections and hyperpigmentation. Although wound healing has been extensively researched, there are gaps in our understanding of the cellular mechanisms involved in the onset and completion of re-epithelization, leading to a scarcity of available enhanced micro-wound healing compounds (MHCs). To investigate the utility of a panel of putative MHCs, a new high-throughput scratch assay method was devised. A guide for multichannel pipette scratches was produced by three-dimensional printing an autoclavable 96-well plate lid with custom-made lengthwise slits. Using an immortalized keratinocyte cell line (N/TERT), the rate of scratch healing was characterized. Critical re-epithelization time points have been determined, and markers of migration are being explored within the identified periods. Moreover, known keratinocyte migration-inducing compounds are being tested to serve as a positive control for enhanced MHC screening. Any putative MHC hits will be further explored using proteomic approaches to elucidate the signalling pathways involved in the onset and completion of re-epithelization. This analysis will assist in the identification of novel enhanced MHC targets and further our understanding of keratinocyte re-epithelization.
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Orfanoudaki, Maria, Anja Hartmann, Mostafa Alilou, et al. "Absolute Configuration of Mycosporine-Like Amino Acids, Their Wound Healing Properties and In Vitro Anti-Aging Effects." Marine Drugs 18, no. 1 (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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Lee, Wen-Jui. "Abstract 29: Utilizing Human Umbilical Cord Blood-Derived Exosomes for Inflammation Modulation and Tissue Regeneration: A Promising Therapeutic Strategy." Stem Cells Translational Medicine 13, Supplement_1 (2024): S32. http://dx.doi.org/10.1093/stcltm/szae062.029.
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Abstract Introduction Umbilical cord blood (UCB)-derived exosomes, with their cup-shaped morphology and rich cargo of proteins, lipids, and RNAs, are small extracellular vesicles isolated from the umbilical cord blood. Notably, they express specific markers such as CD9, CD63, and CD81. UCB-derived exosomes have shown remarkable potential in anti-inflammatory responses and tissue regeneration, making them a hope for therapeutic applications in various medical fields. Objectives This study aims to evaluate the effect of UCB-derived exosomes, a potential game-changer, on exploring potential future applications of these exosomes in regenerative medicine, immunotherapy, and other therapeutic areas. Methods The size distribution and concentration of exosomes were analyzed using nanoparticle tracking analysis (NTA). A scratch wound healing assay was used to measure wound coverage at 0 and 10 hours, both in the presence and absence of exosomes. The concentration of Tumor Necrosis Factor (TNF) was measured using an ELISA assay in control and exosome-treated samples under different conditions to assess the immunomodulatory properties of the exosomes. Cell proliferation was monitored over five days using the MTT assay to determine the proliferative effect of exosomes on cell growth. Results NTA was conducted to identify the purified nanoparticles derived from UCB. The isolated exosomes exhibited a spherical morphology with an average size of approximately 115.4 nm and expressed exosome-specific markers CD9, CD63, and CD81. The results demonstrated that all UCB-derived exosomes significantly reduced TNF expression in LPS-challenged macrophages. Subsequently, we incubated CCD966SK cells with UCB-derived exosomes. MTT and scratch wound assays showed that UCB-derived exosomes significantly promoted cell proliferation and migration. Discussion The study's findings, which highlight the significant therapeutic potential of exosomes in enhancing wound healing, modulating immune responses, and promoting cell proliferation, could potentially revolutionize regenerative medicine and immunotherapy. The wound healing assay results confirm that exosomes facilitate cellular repair processes. Immunomodulation assays showed that exosomes can effectively alter immune responses, which is potentially beneficial for treating inflammatory conditions. The increased cell growth observed in the MTT assay suggests that exosomes may support tissue regeneration and repair. These findings underscore the promising role of exosomes in therapeutic interventions, offering new avenues for treatment and healing.
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Amin, Zahra A., Hapipah M. Ali, Mohammed A. Alshawsh, Pouya H. Darvish, and Mahmood A. Abdulla. "Application ofAntrodia camphorataPromotes Rat’s Wound HealingIn Vivoand Facilitates Fibroblast Cell ProliferationIn Vitro." Evidence-Based Complementary and Alternative Medicine 2015 (2015): 1–14. http://dx.doi.org/10.1155/2015/317693.
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Antrodia camphoratais a parasitic fungus from Taiwan, it has been documented to possess a variety of pharmacological and biological activities. The present study was undertaken to evaluate the potential ofAntrodia camphorataethanol extract to accelerate the rate of wound healing closure and histology of wound area in experimental rats. The safety ofAntrodia camphoratawas determinedin vivoby the acute toxicity test andin vitroby fibroblast cell proliferation assay. The scratch assay was used to evaluate thein vitrowound healing in fibroblast cells and the excision model of wound healing was testedin vivousing four groups of adultSprague Dawleyrats. Our results showed that wound treated withAntrodia camphorataextract and intrasite gel significantly accelerates the rate of wound healing closure than those treated with the vehicle. Wounds dressed withAntrodia camphorataextract showed remarkably less scar width at wound closure and granulation tissue contained less inflammatory cell and more fibroblast compared to wounds treated with the vehicle. Masson’s trichrom stain showed granulation tissue containing more collagen and less inflammatory cell inAntrodia camphoratatreated wounds. In conclusion,Antrodia camphorataextract significantly enhanced the rate of the wound enclosure in rats and promotes thein vitrohealing through fibroblast cell proliferation.
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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 (2011): 158–60. http://dx.doi.org/10.1016/j.ab.2010.12.012.
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Parabucki, Ana, Anja Santrač, Danijela Savić, et al. "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 (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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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 (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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Martinotti, Simona, Giorgia Pellavio, Umberto Laforenza, and Elia Ranzato. "Propolis Induces AQP3 Expression: A Possible Way of Action in Wound Healing." Molecules 24, no. 8 (2019): 1544. http://dx.doi.org/10.3390/molecules24081544.
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Propolis is the generic name of a complex of resinous compound collected by honeybees and it has been utilized for many years in folk medicine. As other products generated by honeybees (such as royal jelly, pollen, honey), propolis has great therapeutic properties, but very little scientific information is available. Therefore, this study was aimed at exploring the potential wound healing properties of propolis. To that end, we utilized an in vitro scratch wound healing model consisting of human immortalized keratinocytes. Our scratch wound data clearly demonstrated that propolis induced a pronounced increase in the wound repair abilities of keratinocytes. A cell migration assay showed that propolis stimulated keratinocytes to close the wound. We revealed the role of H2O2 as the main mediator of propolis regenerative properties. We showed that this extracellularly released H2O2 could pass across the plasma membrane through a specific aquaporin (i.e., AQP3) modulating intracellular responses. The data offer a biological characterization of propolis positive effects suggesting that propolis could also be utilized in wound treatment within clinical settings.
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Yue, Patrick Y. K., Emily P. Y. Leung, N. K. Mak, and Ricky N. S. Wong. "A Simplified Method for Quantifying Cell Migration/Wound Healing in 96-Well Plates." Journal of Biomolecular Screening 15, no. 4 (2010): 427–33. http://dx.doi.org/10.1177/1087057110361772.
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Cell migration plays a key role in both normal physiological and pathological conditions. The study of cell migration and its underlying mechanisms is of great significance in various fields of research, including basic biology and pharmaceutical development. The cell migration or scratch wounding assay is an easy and economical in vitro method that allows researchers to assess a large number of testing compounds. Even though this simple assay has been used for decades, researchers are still trying to modify such experimental protocols and wounding devices. In this study, an 8-channel mechanical “wounder” was designed for performing a cell migration assay, particularly in a 96-well culture plate format. With special designs of a guiding bar and adjustable pins for use with disposable pipette tips, this wounder confined the scratch area within the center of each well to ensure a perfect contact between the pins and the well surface. As a result, this mechanical wounder produces a uniform denudation of a cell monolayer in a 96-well plate with a wound size of around 600 µm. Using this improved wounding device, the effects of epidermal growth factor and DL-α-difluoromethylornithine on the reepithelialization of rat intestinal epithelial cells (IEC-6) and serum on the wound recovery of human umbilical vein endothelial cells were demonstrated. This wounder facilitates cell migration study and can be applicable for multiple sample analysis.
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Greene-Roos, Jennifer A., and Mary Laughlin. "Umbilical cord derived monocytes and platelet rich plasma for diabetic wound healing." Journal of Immunology 198, no. 1_Supplement (2017): 81.27. http://dx.doi.org/10.4049/jimmunol.198.supp.81.27.
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Abstract Chronic non-healing ulcers are a significant complication present in approximately 15% of diabetic patients. Despite standard clinical treatment including wound dressing, debridement of necrotic tissue, and offloading, 15–27% of patients with diabetic ulcer will require amputation. Deep infection is a risk factor for amputation. Platelet rich plasma (PRP) and stem cells have been tested for potential application to treat diabetic ulcers and have been shown to improve wound healing. Umbilical cord blood provides robust cellular and secreted products with therapeutic potential, but remains yet to be fully exploited as a resource for wound healing strategies. Our initial proof of concept studies indicate that umbilical cord blood derived monocytes and platelet rich plasma enhance wound healing processes measured by in vitro assays. When compared to monocytes or platelet rich plasma alone, monocytes and platelet rich plasma significantly enhanced neovascularization as measured by the matrigel angiogenesis assay. Monocytes and platelet rich plasma enhanced fibroblast migration in a wound scratch assay. Additionally, monocytes and platelet rich plasma enhance proliferation of endothelial cells and fibroblasts. Furthermore, these monocytes have potent antibacterial effects as measured by their ability to phagocytose P. aeruginosa in vitro. Collectively, this in vitro data indicates that this product could heal wounds by a mechanism involving enhancing neovascularization, proliferation, migration, and antibacterial activities, all processes which are dysregulated in diabetic wound healing.
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Ariesanti, Yessy, Ferry Sandra, Bianda Claresta, and Livia Alvita. "Coffea canephora Bean Extract Induces NIH3T3 Cell Migration." Indonesian Biomedical Journal 13, no. 2 (2021): 216–20. http://dx.doi.org/10.18585/inabj.v13i2.1522.
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BACKGROUND: Wound healing is an essential biological process that consists of sequential steps aimed at restoring the architecture and function of damaged cells and tissues. There are empirical evidences of using pure coffee bean powder as an alternative medicine in treating various types of wounds. However, there is limited data on coffee-induced wound healing, especially migration of cells. Therefore, current study was conducted to investigate the role of coffee extract in cell migration, especially fibroblast which is important for wound healing.METHODS: Coffea canephora beans were prepared, extracted and added in the NIH3T3 cell culture in final concentration of 2.5% and 5%. Then cytotoxicity test was performed using Na,30-[(phenylamino)-carbonyl]-3,4-tetrazolium]-bis(4-methoxy-6-nitro) benzenesulfonic acid hydrate (XTT) assay. Meanwhile, cell migration test was performed with scratch assay. All results were statistically analyzed.RESULTS: The 2.5% or 5% Coffea canephora beans extract (CCBE)-treated NIH3T3 cell numbers were almost similar with the numbers of NIH3T3 cells in starvation medium merely. Meanwhile, 2.5% and 5% CCBE showed significant decrease of the widths of scratched areas compared to starvation medium merely (ANOVA with LSD Post-hoc, p=0.000). After 24 h and 48 h, the average widths of 2.5% and 5% CCBE-treated scratched areas were 235.68±22.79, 50.36±5.29, 229.95±23.01, 27.68±2.83, respectively.CONCLUSION: Since both 2.5% and 5% CCBE are potential in inducing migration of fibroblast (NIH3T3 cell) and do not induce cytotoxicity, the CCBE could be potential as an agent for wound healing.KEYWORDS: coffee, Coffea canephora, NIH3T3, migration, cytotoxicity
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Mohammad Ali, Sara Torabi, and Farid Semsarha. "An In Vitro Investigation of T-Consciousness Fields' Effects on Wound Healing through Scratch Assay." Scientific Journal of Cosmointel 4, no. 16 (2025): 26–33. https://doi.org/10.61450/joci.v4i16.205.
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Various theories about Consciousness have been proposed, but there is no consensus definition within the scientific community. Taheri introduced the concept of T-Consciousness to differentiate his perspective from others. According to his theory, T-Consciousness is a non-physical entity and serves as the fundamental element of the universe, from which information, matter, and energy originate. This theory also posits the existence of various T-Consciousness Fields (TCFs), each with distinct functions, that are the subcategory of the Cosmic Consciousness Network (CCN), representing the whole consciousness of the universe. In the current experiment, we investigated the wound-healing effects of two types of TCFs: the Faradarmani Consciousness Field (FCF) and the T-Consciousness Charge Field (TCCF), using a scratch assay. According to Taheri’s theory, when a sample is exposed to TCFs, the information transmitted can alter the properties of that substance. To test this, we used endometrial stem cells (EnSC) as a model for in vitro wound healing. The cells were mechanically wounded with a sterile pipette tip, and the wound closure rate was measured by capturing images at 0, 6, 12, and 24-hour intervals. The experiment included four groups: control (0), control with water, cells exposed to FCF-treated water, and cells exposed to TCCF-treated water. After 6 hours, a significant reduction in empty space was observed in the TCCF-treated group compared to the control 0 group (P-value < 0.05), indicating an acceleration in wound healing. Additionally, after 24 hours, there was an insignificant 6% improvement in healing in the water control group compared to the control (0) group, suggesting that water alone facilitates healing processes. However, treatment with TCCF-treated water and FCF-treated water resulted in approximately 56% and 33% healing, respectively, compared to the control (0) group, and a 47% and 26% improvement in wound repair compared to the water control group (P-value < 0.001). On average, TCCF demonstrated a 75% greater effectiveness in promoting wound healing compared to FCF. Further studies are needed to understand the molecular mechanisms through which TCFs influence cellular processes and wound repair.
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Husni, Elidahanum, Dachriyanus Hamidi, Dean Pavvellin, Hafizatul Hidayah, and Suryati Syafri. "Metabolite profiling, antioxidant, and in vitro wound healing activities of Citrus medica L. and Citrus x microcarpa Bunge peels and leaves essential oils." Prospects in Pharmaceutical Sciences 22, no. 4 (2024): 122–30. https://doi.org/10.56782/pps.250.
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Wound care is still dominated by the use of synthetic chemicals such as anti-inflammatory drugs, corticosteroids and antibiotics which have many shortcomings. One potential alternative to be developed is essential oils. Citrus are members of the Rutaceae family which contain essential oils. Citrus medica L. and Citrus x microcarpa Bunge are distributed in West Sumatra and research on wound healing activity is still lacking. Therefore, this study aims to identify the metabolite profiles of essential oils of two species of citrus using FTIR spectroscopy and GC-MS as well as evaluate their antioxidant activity and determine their wound healing activity. Essential oils were obtained by the hydrodistillation method. Metabolite fingerprinting and profiling were carried out by using FTIR and GC-MS methods. Antioxidant activity was determined using the ABTS and FRAP methods while wound healing used the MTT Assay and Scratch Assay. The result showed the secondary metabolites of Citrus medica peels (LPEO) and Citrus medica leaves (LLEO) essential oil contained D-limonene and citral. Meanwhile Citrus x microcarpa leaves (MLEO) contain β-pinene, germacrene D, and elemol and Citrus x microcarpa peels essential oil (MPEO) consists of D-limonene, β-pinene, and germacrene D. MLEO has stronger antioxidant activity than MPEO, LPEO and LPEO with ABTS and FRAP method with IC50 197.051 µg/mL and 8.04 Fe (II)/mg sample and is followed by LLEO. Testing of wound healing activity using MTT assay showed that MPEO significantly increased cell proliferation. Meanwhile, the highest wound closure was found at a concentration of 0.1 µg/mL for MLEO followed by LPEO at 77.54% and 73.71% using scratch assay method. It can be concluded that essential oils of Citrus medica and Citrus x microcarpa can increase cell proliferation and migration so that they have potential to be developed as an alternative in wound healing.