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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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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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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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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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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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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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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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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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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 (June 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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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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Cheng, Yu, Zhang Hu, Yuntao Zhao, Zuhao Zou, Sitong Lu, Bijun Zhang, and Sidong Li. "Sponges of Carboxymethyl Chitosan Grafted with Collagen Peptides for Wound Healing." International Journal of Molecular Sciences 20, no. 16 (August 9, 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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Shabestani Monfared, Ghazal, Peter Ertl, and Mario Rothbauer. "Microfluidic and Lab-on-a-Chip Systems for Cutaneous Wound Healing Studies." Pharmaceutics 13, no. 6 (May 26, 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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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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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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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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Kawano, Yayoi, Viorica Patrulea, Emmanuelle Sublet, Gerrit Borchard, Takuya Iyoda, Rihoko Kageyama, Asa Morita, et al. "Wound Healing Promotion by Hyaluronic Acid: Effect of Molecular Weight on Gene Expression and In Vivo Wound Closure." Pharmaceuticals 14, no. 4 (March 28, 2021): 301. http://dx.doi.org/10.3390/ph14040301.
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Hyaluronic acid (HA) has been known to play an important role in wound healing process. However, the effect of molecular weight (MW) of exogenously administered HA on the wound healing process has not been fully understood. In this study, we investigated HA with different MWs on wound healing process using human epidermal keratinocytes and dermal fibroblasts. Cell proliferation and migration ability were assessed by water soluble tetrazolium (WST) assay and wound scratch assay. We examined the effect of HA addition in a full-thickness wound model in mice and the gene expression related to wound healing. Proliferation and migration of HaCaT cells increased with the increase of MW and concentration of HA. Interleukin (IL-1β), IL-8 and vascular endothelial growth factor (VEGF) as well as matrix metalloproteinase (MMP)-9 and MMP-13 were significantly upregulated by high molecular weight (HMW) HA in keratinocytes. Together with VEGF upregulation and the observed promotion of HaCaT migration, HA with the MW of 2290 kDa may hold potential to improve re-epithelialization, a critical obstacle to heal chronic wounds.
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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 (April 2011): 158–60. http://dx.doi.org/10.1016/j.ab.2010.12.012.
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Yıldız, Erdost, Özgün Melike Gedar Totuk, Adriano Mollica, Kerem Kabadayı, and Afsun Şahin. "Effects of Biphalin on Corneal Epithelial Wound Healing." Proceedings 2, no. 25 (December 5, 2018): 1552. http://dx.doi.org/10.3390/proceedings2251552.
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After physical or surgical damage of corneal epithelium, most of analgesic drugs, like non-selective opioid agonists and non-steroid anti-inflammatory drugs, cannot be used because of their negative effects on wound healing process. Biphalin is selective µ and Δ opioid receptor agonist which has proven analgesic effects on rodents. Our purpose of study is finding effects of biphalin on wound healing of corneal epithelium. We used primary culture of human corneal epithelial cells (HCECs) for examining effects of biphalin on wound healing. Firstly, we measured toxicity of Biphalin in various concentrations with MTT assay and we showed biphalin has no toxic effects on HCECs in lower concentrations than 100 µM in various incubation times. After MTT assay, we administered 1 µM and 10 µM biphalin at in vitro scratch assay of HCECs, biphalin increased wound closure process significantly at 1 µM concentration (p < 0.05). Then we tested effects of biphalin on cell migration and proliferation separately. Bifalin increased migration of HCECs significantly (p < 0.01) at transwell migration assay. But we did not observe any significant difference between groups in Ki67 proliferation assay. In all these experiments, we also used naloxone to inhibiting effects of biphalin. In biphalin plus naloxone groups, effects of biphalin decrease partially. Our study results suggest, biphalin has positive effects on epithelial wound healing via opioid receptors. This effect because of increased migration of HCECs under influence of biphalin. With these findings, we propose biphalin as a new analgesic agent for post-surgical and post-traumatic care of corneal epithelial wounds.
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Boran, Rukiye, Nurdan Sarac, Tuba Baygar, and Aysel Ugur. "The Role of Hypericum Lydium Boiss. (Hypericaceae) in Soft Tissue Healing and its Capacity to Prevent Infections after Dental Extraction." Alinteri Journal of Agriculture Sciences 36, no. 1 (May 17, 2021): 226–33. http://dx.doi.org/10.47059/alinteri/v36i1/ajas21033.
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The genus Hypericum sp. has a number of uses in traditional medicine like curing the burns, ulcers, haemorrhoids and wound healing. The species Hypericum lydium Boiss. (Hypericaceae), however, has not been known to have any properties related to the healing of injuries or antimicrobial working against the oral microorganisms. The present study was aimed to evaluate the efficiency of H. lydium in soft tissue healing and its capacity to prevent infections after dental extraction. H. lydium was extracted with ethanol and the obtained extract was tested for its inhibition ability on extracellular matrix-degrading enzymes; collagenase, hyaluronidase and elastase. To determine the cytotoxicity and wound healing capacity of the extract, MTT and in vitro scratch wound healing assay were performed using the NIH-3T3 fibroblast cells, respectively. Antimicrobial activity was investigated by microdilution method against oral pathogenic microorganisms. The highest enzyme inhibition activity was determined against elastase (80.27±0.1%). According to the cytotoxic activity results, the IC50 value of the H. lydium was found to be 82.20±4.05 μg/mL. Scratch wound healing assay of the extract exhibited a significant enhancement at 24 h with a closed wound area when compared with the control. The extract showed potent antimicrobial properties against oral pathogenic microorganisms. The results of the study revealed out that H. lydium can be considered as a natural compound for dental industry to improve soft tissue healing and to prevent the possible infections after dental extraction.
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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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Jose, Deepak Tom, Sivagurunathan, P, Aswini, B, Dinesh, MD, and Uma, C. "Development of Antimicrobial Conjugates and Evaluating its Antibacterial potential and Wound Healing ability." Journal of University of Shanghai for Science and Technology 23, no. 09 (September 13, 2021): 540–55. http://dx.doi.org/10.51201/jusst/21/09536.
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Antimicrobial peptides from Streptomyces sp. and marine fish (Carangoides malabaricus) were extracted and developed as conjugates in the present study. The objective was framed to analyze the ability of conjugate to retard the growth of test bacteria causing diabetic foot ulcers. Fibroblast cell adhesion on AMP conjugates coated mesh samples were recorded using microscopic studies with an aim of developing a novel tissue engineered wound dressing material. Thus developed tissue engineered materials were evaluated for its antibacterial potential against wound pathogens; and to assay the wound healing ability using a standard in vitro wound scratch method. Tissue engineered materials were developed using L929 fibroblast cells. L929 fibroblast cells attachment and its stage wise development on wound dressing mesh materials were microscopically observed. In vitro wound healing assay revealed that the developed conjugates (containing AMPs) exhibited cell migration and proliferation after 12th hour of incubation indicating the wound healing abilities. The results showed that the developed tissue engineered wound dressing material has commercial interest in near future.
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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 (April 19, 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 (March 5, 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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Harikarnpakdee, Saraporn, and Verisa Chowjarean. "Grammatophyllum speciosum Ethanolic Extract Promotes Wound Healing in Human Primary Fibroblast Cells." International Journal of Cell Biology 2018 (October 21, 2018): 1–6. http://dx.doi.org/10.1155/2018/7836869.
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Grammatophyllum speciosum is a plant in Orchidaceae family which contains a variety of phytochemical compounds that might be beneficial for medicinal use. This study aimed to evaluate the activity of pseudobulb of G. speciosum extract (GSE) in wound healing processes in human primary fibroblast cells along with in vitro antioxidant activity and total phenolic content of GSE. Scratch wound healing assay indicated that GSE was capable of increasing migration rate after 6 and 9 hours of treatment. Besides, the extract was able to scavenge DPPH, ABTS, and superoxide anion radicals indicating the antioxidative property of GSE. This study suggested a novel role of the of pseudobulb extract of G. speciosum as a wound healing enhancer. The results from this study might be beneficial for the development of further novel active compounds for skin wound healing.
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Kim, JaeGoo, Yu-Kyong Shin, and Ki-Young Kim. "Promotion of Keratinocyte Proliferation by Tracheloside through ERK1/2 Stimulation." Evidence-Based Complementary and Alternative Medicine 2018 (July 26, 2018): 1–5. http://dx.doi.org/10.1155/2018/4580627.
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Cell migration and proliferation are important for proper wound healing after skin injury. Recent studies have shown that compounds from plants could promote cell migration and proliferation. Tracheloside, which is a plant lignan, has been found to promote the growth of HaCaT cells over 40% compared to other compounds tested based on a cell proliferation assay. An in vitro scratch assay confirmed the healing activity of tracheloside (more than 2-fold increased healing activity after 24 hours of treatment compared with the control) and revealed that this activity is better than that of allantoin (1.2-fold increased after 24 hours of treatment compared with the control), a positive control. With western blot results, wound healing with tracheloside occurred through the phosphorylation of ERK1/2. Therefore, tracheloside is a good candidate to promote wound healing and could be developed as a therapeutic agent for wound treatment or used as a leading compound with higher activity.
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Vaid, Bhavna, Bhupinder Singh Chopra, Sachin Raut, Amin Sagar, Maulik D. Badmalia, Ashish, and Neeraj Khatri. "Antioxidant and Wound Healing Property of Gelsolin in 3T3-L1 Cells." Oxidative Medicine and Cellular Longevity 2020 (February 12, 2020): 1–7. http://dx.doi.org/10.1155/2020/4045365.
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Delineation of factors which affect wound healing would be of immense value to enable on-time or early healing and reduce comorbidities associated with infections or biochemical stress like diabetes. Plasma gelsolin has been identified earlier to significantly enable injury recovery compared to placebo. This study evaluates the role of rhuGSN for its antioxidant and wound healing properties in murine fibroblasts (3T3-L1 cell line). Total antioxidant capacity of rhuGSN increased in a concentration-dependent manner (0.75-200 μg/mL). Cells pretreated with 0.375 and 0.75 μg/mL rhuGSN for 24 h exhibited a significant increase in viability in a MTT assay. Preincubation of cells with rhuGSN for 24 h followed by oxidative stress induced by exposure to H2O2 for 3 h showed cytoprotective effect. rhuGSN at 12.5 and 25 μg/mL concentration showed an enhanced cell migration after 20 h of injury in a scratch wound healing assay. The proinflammatory cytokine IL-6 levels were elevated in the culture supernatant. These results establish an effective role of rhuGSN against oxidative stress induced by H2O2 and in wound healing of 3T3-L1 fibroblast cells.
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Arranz-Valsero, Isabel, Laura Soriano-Romaní, Laura García-Posadas, Antonio López-García, and Yolanda Diebold. "IL-6 as a corneal wound healing mediator in an in vitro scratch assay." Experimental Eye Research 125 (August 2014): 183–92. http://dx.doi.org/10.1016/j.exer.2014.06.012.
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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 (August 2, 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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Bagheri, Elham, Kamelia Saremi, Fatemeh Hajiaghaalipour, Fadhil Lafta Faraj, Hapipah Mohd Ali, Mahmood Ameen Abdulla, Si Lay Khaing, and Nur`Ain Salehen. "Synthesis of Novel Derivatives of Quinazoline Schiff base Compound Promotes Epithelial Wound Healing." Current Pharmaceutical Design 24, no. 13 (July 11, 2018): 1395–404. http://dx.doi.org/10.2174/1381612824666180130124308.
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Quinazoline is an aromatic bicyclic compound exhibiting several pharmaceutical and biological activities. This study was conducted to investigate the potential wound healing properties of Synthetic Quinazoline Compound (SQC) on experimental rats. The toxicity of SQC was determined by MTT cell proliferation assay. The healing effect of SQC was assessed by in vitro wound healing scratch assay on the skin fibroblast cells (BJ-5ta) and in vivo wound healing experiment of low and high dose of SQC on adult Sprague-Dawley rats compared with negative (gum acacia) and positive control (Intrasite-gel). Hematoxylin and Eosin (H&E), Masson’s Trichrome (MT) staining and immunohistochemistry analysis were performed to evaluate the histopathological alterations and proteins expression of Bax and Hsp70 on the wound tissue after 10 days. In addition, levels of antioxidant enzymes (catalase, glutathione peroxidase and superoxide dismutase), and malondialdehyde (MDA) were measured in wound tissue homogenates. The SQC significantly enhanced BJ-5ta cell proliferation and accelerated the percentage of wound closure, with less scarring, increased fibroblast and collagen fibers and less inflammatory cells compared with the negative control. The compound also increases endogenous enzymes and decline lipid peroxidation in wound homogenate.
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Chaitrakoonthong, Tatcha, Ruchanee Ampornaramveth, and Paksinee Kamolratanakul. "Rinsing with L-Ascorbic Acid Exhibits Concentration-Dependent Effects on Human Gingival Fibroblast In Vitro Wound Healing Behavior." International Journal of Dentistry 2020 (March 21, 2020): 1–7. http://dx.doi.org/10.1155/2020/4706418.
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Vitamin C or L-ascorbic acid has diverse functions in the body, especially healing promotion in tissue injury via participating in the hydroxylation reactions required for collagen formation. Systemic administration of vitamin C plays an important role on gingival fibroblast proliferation and functions. Whether local or rinsing administration of vitamin C alters gingival fibroblast wound healing behavior remains unclear. The aim of this study was to investigate the rinsing effect of vitamin C on gingival fibroblast behavior utilizing an in vitro wound healing model. Primary human gingival fibroblasts isolated from gingival tissue were rinsed with medium containing various concentrations of vitamin C. The rinsing effect of vitamin C on in vitro wound healing was assessed using a scratch test assay. Cell migration, cell viability, and extracellular matrix gene expression were analyzed by transwell migration assay, MTT assay, and real-time RT-PCR, respectively. We found that rinsing with 10 or 20 µg/ml vitamin C significantly increased fibroblast migration (p≤0.05). However, no significant effect was found in the cell viability or in vitro wound healing assays. In contrast, rinsing with 50 µg/ml vitamin C significantly delayed wound closure (p≤0.05). Real-time PCR demonstrated that 50 µg/ml vitamin C significantly increased fibroblast expression of COL1, FN, IL-6, and bFGF. The data demonstrate that rinsing with vitamin C (10/20 µg/ml) accelerates fibroblast migration. However, 50 µg/ml of vitamin C increases the expression of COL1, FN, IL-6, and bFGF, which are related to fibroblast wound healing activity. Prescribing vitamin C with the appropriate duration and drug administration method should be determined to maximize its benefit.
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Fouché, Morné, Clarissa Willers, Sias Hamman, Christiaan Malherbe, and Jan Steenekamp. "Wound Healing Effects of Aloe muth-muth: In Vitro Investigations Using Immortalized Human Keratinocytes (HaCaT)." Biology 9, no. 11 (October 23, 2020): 350. http://dx.doi.org/10.3390/biology9110350.
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The traditional use of Aloe spp. for the purpose of wound healing has a long history and is widespread internationally. Recently, a hybrid aloe plant (Aloe muth-muth) has been cultivated by cross pollination between Aloe vera and Aloe ferox. The Aloe muth-muth plant has not yet been investigated for medicinal properties and provides an opportunity for potential biological activity, including wound healing. The aim of this study was to investigate the in vitro wound healing effects of both Aloe muth-muth gel and whole leaf material with the use of the immortalized human keratinocyte (HaCaT) cell line. Cell viability was conducted using methyl thiazolyl tetrazolium (MTT) assays. In vitro wound healing was tested on HaCaT cells using an established scratch assay method. The effect of Aloe muth-muth gel material on HaCaT cell migration was also investigated. Aloe muth-muth gel material exhibited statistically significantly (p < 0.05) higher percentage wound closure compared to the control at all three concentrations investigated. These findings confirm that this newly cultivated species, Aloe muth-muth, also possesses wound healing activity corresponding to that reported for the two species it is derived from, namely, Aloe vera and Aloe ferox. Therefore, Aloe muth-muth has the potential to be used in future wound therapeutics.
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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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Chmielowiec, Jolanta, Malgorzata Borowiak, Markus Morkel, Theresia Stradal, Barbara Munz, Sabine Werner, Jürgen Wehland, Carmen Birchmeier, and Walter Birchmeier. "c-Met is essential for wound healing in the skin." Journal of Cell Biology 177, no. 1 (April 2, 2007): 151–62. http://dx.doi.org/10.1083/jcb.200701086.
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Wound healing of the skin is a crucial regenerative process in adult mammals. We examined wound healing in conditional mutant mice, in which the c-Met gene that encodes the receptor of hepatocyte growth factor/scatter factor was mutated in the epidermis by cre recombinase. c-Met–deficient keratinocytes were unable to contribute to the reepithelialization of skin wounds. In conditional c-Met mutant mice, wound closure was slightly attenuated, but occurred exclusively by a few (5%) keratinocytes that had escaped recombination. This demonstrates that the wound process selected and amplified residual cells that express a functional c-Met receptor. We also cultured primary keratinocytes from the skin of conditional c-Met mutant mice and examined them in scratch wound assays. Again, closure of scratch wounds occurred by the few remaining c-Met–positive cells. Our data show that c-Met signaling not only controls cell growth and migration during embryogenesis but is also essential for the generation of the hyperproliferative epithelium in skin wounds, and thus for a fundamental regenerative process in the adult.
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Rajalekshmy, GP, and MR Rekha. "Synthesis and evaluation of an alginate-methacrylate xerogel for insulin delivery towards wound healing applications." Therapeutic Delivery 12, no. 3 (March 2021): 215–34. http://dx.doi.org/10.4155/tde-2020-0128.
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Background: Alginate is one of the most widely used biopolymer for wound healing. But poor mechanical strength and degradability limits its application especially as a drug-delivery matrix. The aim of this study was to develop stable alginate based scaffold for insulin delivery toward wound care. Materials & methods: The xerogel alginate-g-poly (methacrylic acid; AGM2S) was characterized by various analytical techniques. Results: AGM2S xerogel showed improved physical stability, low degradation, good swelling and water vapour transmission rate (WVTR). About 70% of insulin was released from loaded xerogel over a period of 48 h and favorably modulated the healing response in in vitro scratch wound assay. Conclusion: Grafting improved the strength and stability of alginate xerogel and the results suggest the application of insulin loaded AGM2S xerogels as a potential wound healing material.
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M. Afonso, Alexandra, Joana Gonçalves, Ângelo Luís, Eugenia Gallardo, and Ana Paula Duarte. "Evaluation of the In Vitro Wound-Healing Activity and Phytochemical Characterization of Propolis and Honey." Applied Sciences 10, no. 5 (March 7, 2020): 1845. http://dx.doi.org/10.3390/app10051845.
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Honey and propolis are natural substances produced by Apis mellifera that contain flavonoids, phenolic acids, and several other phytochemicals. The aim of this study was to phytochemically characterize three different types of honey and propolis, both separately and mixed, and to evaluate their wound-healing activity. Total phenolic compounds and flavonoids were determined using the Folin–Ciocalteu’s and aluminum chloride colorimetric methods, respectively. The antioxidant activity was evaluated by both the DPPH free radical scavenging assay and β-carotene bleaching test, and the anti-inflammatory activity was determined by a protein denaturation method. To evaluate the wound-healing activity of the samples, NHDF cells were subjected to a wound scratch assay. The obtained results showed that dark-brown honey presents a higher concentration of phenolic compounds and flavonoids, as well as higher antioxidant and anti-inflammatory activities. Propolis samples had the highest concentrations in bioactive compounds. Examining the microscopic images, it was possible to verify that the samples promote cell migration, demonstrating the wound-healing potential of honey and propolis.
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Dinh, Han Van. "ID3015 Mesenchymal stem cells for treatment of chronic wounds: A Study with autologous transplantation of adipose-derived stem cells." Biomedical Research and Therapy 4, S (September 5, 2017): 27. http://dx.doi.org/10.15419/bmrat.v4is.236.
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Objective: This study was to determine the effects of adipose-derived stem cells (ADSCs) on dermal fibroblasts responses to injury including migration and proliferation in vitro. We also evaluated the autologous transplantation of ADSCs on treatment of human chronic wounds.
Subjects and methods: The proliferation and migration of fibroblast was evaluated by co-culture ADSCs with allogenic dermal fibroblast and by the scratch assay. In clinical study, autologous ADSCs were transplanted on to chronic wounds of 25 patients, who were hospitalized into the Wound Healing Department of the National Institute of Burns from April, 2015 to June, 2016. The mean age was 56.88 ± 16.81, male/female ratio was 2.12. The autologous adipose-derived stem cells at passages 5 were transplanted on surface of wound every 3÷5 days. The wound biopsies for H&E staining and for Transmission Electron Microscope were taken before transplantation and at day 7, day15 and day 20 of studied progress.
Results: ADSCs stimulated fibroblast proliferation and migration in wound healing assay. In clinical study, before transplantation, extracellular matrix (ECM) was destroyed. After transplantation, ADSCs strongly stimulated fibroblast proliferation and fibroblasts to produce collagen. ADSCs also promoted proliferations of epithelial cells and neovascularization at the chronic wound site.
Conclusion: Autologous ADSCs promoted the wound healing process by cell proliferation and improvement of ECM in chronic wound site.
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Storm, Tina, Iain Wilson, Ross Campbell, Arantxa Bolinches-Amorós, Angela J. Russell, Stephen G. Davies, Alun R. Barnard, and Robert E. MacLaren. "A Semiautomated, Phenotypic, In Vitro Scratch Assay for Assessing Retinal Pigment Epithelial Cell Wound Healing." Journal of Ocular Pharmacology and Therapeutics 36, no. 4 (May 1, 2020): 257–66. http://dx.doi.org/10.1089/jop.2019.0116.
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Ghahremani, Hossein, Majid Sirati-Sabet, and Siamak Salami. "Evaluation of Impacts of Cellular Metabolism on the Migration of Ovarian Cancer Cells by Two in Vitro Assays: A Method Comparison Study." Galen Medical Journal 9 (August 16, 2020): 1831. http://dx.doi.org/10.31661/gmj.v9i0.1831.
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Background: Alteration of metabolic pathways in cancer cells can intensely modulate their migration as an important step in invasion and metastasis. Ketogenic diet showed some contradictory results in cancer patients. In this study the impact of metabolic reprogramming of A2780CP as a model of ovarian cancer stem-like cells on cell migration by two in vitro methods: wound healing and soft agar colony-forming assays. Materials and Methods: short term and long term metabolic reprogramming were done by restriction of glucose to 250mg/L with or without enrichment with beta-hydroxybutyrate (5 milimolar) for 48 hours and 30 days, respectively. Wound healing assay was done and the wound ratio was calculated for 24 and 48 hours. Soft agar colony formation assay was also done in treated and control cells. For method comparison, ten biological replicates were analyzed in triplicate. Results: Migration of A2780CP ovarian cancer stem-like cells were significantly alleviated by long term glucose restriction but no significant changes were observed in short term study. Beta-hydroxybutyrate enrichment did not produce significant impacts on glucose restriction in short or long term studies. Conclusion: The results of colony formation in soft agar and wound or scratch healing assay were in good correlation and convergence which could be used interchangeably in the investigation of metabolic reprogramming in cancer cells. [GMJ.2020;9:e1831]
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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 (June 14, 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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Demyanenko, Ilya A., Vlada V. Zakharova, Olga P. Ilyinskaya, Tamara V. Vasilieva, Artem V. Fedorov, Vasily N. Manskikh, Roman A. Zinovkin, et al. "Mitochondria-Targeted Antioxidant SkQ1 Improves Dermal Wound Healing in Genetically Diabetic Mice." Oxidative Medicine and Cellular Longevity 2017 (2017): 1–10. http://dx.doi.org/10.1155/2017/6408278.
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Oxidative stress is widely recognized as an important factor in the delayed wound healing in diabetes. However, the role of mitochondrial reactive oxygen species in this process is unknown. It was assumed that mitochondrial reactive oxygen species are involved in many wound-healing processes in both diabetic humans and animals. We have applied the mitochondria-targeted antioxidant 10-(6′-plastoquinonyl)decyltriphenylphosphonium (SkQ1) to explore the role of mitochondrial reactive oxygen species in the wound healing of genetically diabetic mice. Healing of full-thickness excisional dermal wounds in diabetic C57BL/KsJ-db−/db− mice was significantly enhanced after long-term (12 weeks) administration of SkQ1. SkQ1 accelerated wound closure and stimulated epithelization, granulation tissue formation, and vascularization. On the 7th day after wounding, SkQ1 treatment increased the number of α-smooth muscle actin-positive cells (myofibroblasts), reduced the number of neutrophils, and increased macrophage infiltration. SkQ1 lowered lipid peroxidation level but did not change the level of the circulatory IL-6 and TNF. SkQ1 pretreatment also stimulated cell migration in a scratch-wound assay in vitro under hyperglycemic condition. Thus, a mitochondria-targeted antioxidant normalized both inflammatory and regenerative phases of wound healing in diabetic mice. Our results pointed to nearly all the major steps of wound healing as the target of excessive mitochondrial reactive oxygen species production in type II diabetes.
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Scheda, Riccardo, Silvia Vitali, Enrico Giampieri, Gianni Pagnini, and Isabella Zironi. "Study of Wound Healing Dynamics by Single Pseudo-Particle Tracking in Phase Contrast Images Acquired in Time-Lapse." Entropy 23, no. 3 (February 26, 2021): 284. http://dx.doi.org/10.3390/e23030284.
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Cellular contacts modify the way cells migrate in a cohesive group with respect to a free single cell. The resulting motion is persistent and correlated, with cells’ velocities self-aligning in time. The presence of a dense agglomerate of cells makes the application of single particle tracking techniques to define cells dynamics difficult, especially in the case of phase contrast images. Here, we propose an original pipeline for the analysis of phase contrast images of the wound healing scratch assay acquired in time-lapse, with the aim of extracting single particle trajectories describing the dynamics of the wound closure. In such an approach, the membrane of the cells at the border of the wound is taken as a unicum, i.e., the wound edge, and the dynamics is described by the stochastic motion of an ensemble of points on such a membrane, i.e., pseudo-particles. For each single frame, the pipeline of analysis includes: first, a texture classification for separating the background from the cells and for identifying the wound edge; second, the computation of the coordinates of the ensemble of pseudo-particles, chosen to be uniformly distributed along the length of the wound edge. We show the results of this method applied to a glioma cell line (T98G) performing a wound healing scratch assay without external stimuli. We discuss the efficiency of the method to assess cell motility and possible applications to other experimental layouts, such as single cell motion. The pipeline is developed in the Python language and is available upon request.
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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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Qiu, Shuo, Yachao Jia, Yunchu Sun, Pei Han, Jia Xu, Gen Wen, and Yimin Chai. "Von Hippel-Lindau (VHL) Protein Antagonist VH298 Improves Wound Healing in Streptozotocin-Induced Hyperglycaemic Rats by Activating Hypoxia-Inducible Factor- (HIF-) 1 Signalling." Journal of Diabetes Research 2019 (February 17, 2019): 1–10. http://dx.doi.org/10.1155/2019/1897174.
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Aims. The purpose of the present research is to investigate the effects of the VHL protein antagonist, VH298, on functional activities of fibroblasts and vascular endothelial cells and the effects on the wound healing process in a streptozotocin-induced hyperglycaemic rat model. Methods. HIF-1α and hydroxy-HIF-1α protein levels in VH298-treated rat fibroblasts (rFb) were measured by immunoblotting, rFb proliferation was detected by the CCK-8 assay, and mRNA levels of related genes were measured by quantitative RT-PCR. In vitro wound healing was simulated by the scratch test; angiogenesis was measured by the human umbilical vein endothelial cell (hUVEC) tube formation assay. VH298 or PBS was locally injected into wounds in rat models with streptozotocin- (STZ-) induced hyperglycaemia, the wound tissues were harvested, and haematoxylin-eosin (HE) and Masson trichrome staining and immunohistochemical processes were conducted. Results. HIF-1α and hydroxy-HIF-1α levels increased in VH298-treated rFb, in a time- and dose-dependent manner. Thirty micromolar VH298 could significantly increase cell proliferation, angiogenesis, and gene expression of type I collagen-α1 (Col1-α1), vascular endothelial growth factor A (VEGF-A), and insulin-like growth factor 1 (IGF-1). The VH298-treated wound had a better healing pattern, activation of HIF-1 signalling, and vascularization. Conclusions. Taken together, VH298 activated the HIF-1 signalling pathway by stabilizing both HIF-1α and hydroxy-HIF-1α. VH298 enhanced rFb functions, promoted hUVEC angiogenesis, and accelerated wound healing in the rat model mimicking diabetes mellitus.
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Iwanabe, Yujiro, Chihiro Masaki, Akiko Tamura, Shintaro Tsuka, Taro Mukaibo, Yusuke Kondo, and Ryuji Hosokawa. "The effect of low-intensity pulsed ultrasound on wound healing using scratch assay in epithelial cells." Journal of Prosthodontic Research 60, no. 4 (October 2016): 308–14. http://dx.doi.org/10.1016/j.jpor.2016.03.002.
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Fronza, M., B. Heinzmann, M. Hamburger, S. Laufer, and I. Merfort. "Determination of the wound healing effect of Calendula extracts using the scratch assay with 3T3 fibroblasts." Journal of Ethnopharmacology 126, no. 3 (December 2009): 463–67. http://dx.doi.org/10.1016/j.jep.2009.09.014.
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Zhao, Ying, Qiang Wang, Yuan Jin, Yadan Li, Changjun Nie, Peipei Huang, Zhixin Li, et al. "Discovery and Characterization of a High-Affinity Small Peptide Ligand, H1, Targeting FGFR2IIIc for Skin Wound Healing." Cellular Physiology and Biochemistry 49, no. 3 (2018): 1074–89. http://dx.doi.org/10.1159/000493287.
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Background/Aims: How to aid recovery from severe skin injuries, such as burns, chronic or radiation ulcers, and trauma, is a critical clinical problem. Current treatment methods remain limited, and the discovery of ideal wound-healing therapeutics has been a focus of research. Functional recombinant proteins such as basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) have been developed for skin repair, however, some disadvantages in their use remain. This study reports the discovery of a novel small peptide targeting fibroblast growth factor receptor 2 IIIc (FGFR2IIIc) as a potential candidate for skin wound healing. Methods: A phage-displayed peptide library was used for biopanning FGFR2IIIc-targeting small peptides. The selected small peptides binding to FGFR2IIIc were qualitatively evaluated by an enzyme-linked immunosorbent assay. Their biological function was detected by a cell proliferation assay. Among them, an optimized small peptide named H1 was selected for further study. The affinity of the H1 peptide and FGFR2IIIc was determined by an isothermal titration calorimetry device. The ability of theH1 peptide to promote skin wound repair was investigated using an endothelial cell tube formation assay and wound healing scratch assay in vitro. Subsequently, the H1 peptide was assessed using a rat skin full-thickness wound model and chorioallantoic membrane (CAM) assays in vivo. To explore its molecular mechanisms, RNA-Seq, quantitative real-time PCR, and western blot assays were performed. Computer molecular simulations were also conducted to analyze the binding model. Results: We identified a novel FGFR2IIIc-targeting small peptide, called H1, with 7 amino acid residues using phage display. H1 had high binding affinity with FGFR2IIIc. The H1 peptide promoted the proliferation and motility of fibroblasts and vascular endothelial cells in vitro. In addition, the H1 peptide enhanced angiogenesis in the chick chorioallantoic membrane and accelerated wound healing in a rat full-thickness wound model in vivo. The H1 peptide activated both the PI3K-AKT and MAPK-ERK1/2 pathways and simultaneously increased the secretion of vascular endothelial growth factor. Computer analysis demonstrated that the model of H1 peptide binding to FGFR2IIIc was similar to that of FGF2 and FGFR2IIIc. Conclusion: The H1 peptide has a high affinity for FGFR2IIIc and shows potential as a wound healing agent. As a substitute for bFGF, it could be developed into a novel therapeutic candidate for skin wound repair in the future.
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Mazutti da Silva, Sandra, Claudio Rezende Costa, Guilherme Martins Gelfuso, Eliete Silva Guerra, Yanna de Medeiros Nóbrega, Sueli Gomes, Aline Pic-Taylor, Yris Fonseca-Bazzo, Damaris Silveira, and Pérola Magalhães. "Wound Healing Effect of Essential Oil Extracted from Eugenia dysenterica DC (Myrtaceae) Leaves." Molecules 24, no. 1 (December 20, 2018): 2. http://dx.doi.org/10.3390/molecules24010002.
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The use of natural oils in topical pharmaceutical preparations has usually presented safe agents for the improvement of human health. Based on research into the immense potential of wound management and healing, we aimed to validate the use of topical natural products by studying the ability of the essential oil of Eugenia dysenterica DC leaves (oEd) to stimulate in vitro skin cell migration. Skin cytotoxicity was evaluated using a fibroblast cell line (L929) by MTT assay. The oil chemical profile was investigated by GC-MS. Moreover, the inhibition of lipopolysaccharide (LPS) induced nitric oxide (NO) production in the macrophage cell line (RAW 264.7) tested. The Chick Chorioallantoic Membrane (CAM) assay was used to evaluate the angiogenic activity and irritating potential of the oil. The oEd induces skin cell migration in a scratch assay at a concentration of 542.2 µg/mL. α-humulene and β-caryophyllene, the major compounds of this oil, as determined by GC-MS, may partly explain the migration effect. The inhibition of nitric oxide by oEd and α-humulene suggested an anti-inflammatory effect. The CAM assay showed that treatment with oEd ≤ 292 µg/mL did not cause skin injury, and that it can promote angiogenesis in vivo. Hence, these results indicate the feasibility of the essential oil of Eugenia dysenterica DC leaves to developed dermatological products capable of helping the body to repair damaged tissue.
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Vs, Subeeksha, Anitha Roy, and Lakshmi T. "THE WOUND HEALING PROPERTY OF THYME OLEORESIN FROM THYMUS VULGARIS L. ON HACAT KERATINOCYTES." Asian Journal of Pharmaceutical and Clinical Research 11, no. 9 (September 7, 2018): 169. http://dx.doi.org/10.22159/ajpcr.2018.v11i9.26987.
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Objective: The objective of the present study was to evaluate the wound healing property of thyme oleoresin using HaCaT keratinocytes.Methods: The effect of thyme oleoresin on cell migratory activity of HaCaT keratinocytes was investigated and analyzed using scratch assay. The HaCaT cell line was obtained from NCCS Pune and maintained in Dulbecco’s Modified Eagle’s Media for the study. The keratinocyte cells (HaCaT) were trypsinized for 30 s and passaged to T25 flasks in complete aseptic environment. The effect of thyme oleoresin on wound closure was determined using a 12-well plate. Dulbecco’s Modified Eagle’s medium with dimethyl sulfoxide was used as control. The effect of thyme oleoresin on wound closure was determined microscopically at 20× magnification using Nikon microscope. The experiment was performed in triplicate. The wound area was photographed and analyzed.Results: Thyme oleoresin at 25 μg/ml and 50 μg/ml has significantly promoted the migration of HaCaT cells, thereby leading to wound closure.Conclusion: The study has proved the wound healing property of thyme oleoresin, and hence, it may be used for wound healing purpose in a natural way.
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Haq, Anika, Suneel Kumar, Yong Mao, Francois Berthiaume, and Bozena Michniak-Kohn. "Thymoquinone-Loaded Polymeric Films and Hydrogels for Bacterial Disinfection and Wound Healing." Biomedicines 8, no. 10 (September 28, 2020): 386. http://dx.doi.org/10.3390/biomedicines8100386.
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The purpose of this study was to synthesize and characterize novel biocompatible topical polymeric film and hydrogel systems that have the potential to deliver the antibacterial agent thymoquinone (TQ) directly to the skin target site to manage the local wound infection and thereby wound healing. The polyvinyl pyrrolidone (PVP) matrix-type films containing TQ were prepared by the solvent casting method. In vitro skin permeation studies on human cadaver skin produced a mean flux of 2.3 µg TQ/cm2/h. Human keratinocyte monolayers subjected to a scratch wound (an in vitro wound healing assay) showed 85% wound closure at day 6 in the TQ group (100 ng/mL TQ) as compared to 50% in the vehicle control group (p = 0.0001). In a zone-of-inhibition (ZOI) assay, TQ-containing films and hydrogels completely wiped out Staphylococcus aureus in 10 cm diameter Tryptic Soy Agar plates while 500 µg/mL gentamicin containing filters gave 10 mm of ZOI. In an ex vivo model, TQ-containing films eradicated bacterial colonization on human cadaver skin. Furthermore, in a full-thickness wound infection model in mice, TQ-containing films showed significant activity in controlling Staphylococcus aureus infection, thereby disinfecting the skin wound. In summary, TQ-containing PVP films and hydrogels developed in this study have the potential to treat and manage wound infections.