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      Aesculus hippocastanum L. Extract Does Not Induce Fibroblast to Myofibroblast Conversion but Increases Extracellular Matrix Production In Vitro Leading to Increased Wound Tensile Strength in Rats

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          Abstract

          The ability of horse chestnut extract (HCE) to induce contraction force in fibroblasts, a process with remarkable significance in skin repair, motivated us to evaluate its wound healing potential in a series of experiments. In the in vitro study of the ability of human dermal fibroblasts to form myofibroblast-like cells was evaluated at the protein level (Western blot and immunofluorescence). The in vivo study was conducted on male Sprague-Dawley rats with inflicted wounds (one open circular and one sutured incision) on their backs. Rats were topically treated with two tested HCE concentrations (0.1% and 1%) or sterile water. The control group remained untreated. The incisions were processed for wound tensile strength (TS) measurement whereas the open wounds were subjected to histological examination. On the in vitro level the HCE extract induced fibronectin-rich extracellular matrix formation, but did not induced α-smooth muscle actin (SMA) expression in dermal fibroblasts. The animal study revealed that HCE increased wound TS and improved collagen organization. In conclusion, the direct comparison of both basic wound models demonstrated that the healing was significantly increased following HCE, thus this extract may be found useful to improve healing of acute wounds. Nevertheless, the use of an experimental rat model warrants a direct extrapolation to the human clinical situation.

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          Most cited references26

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          Keratinocyte-fibroblast interactions in wound healing.

          Cutaneous tissue repair aims at restoring the barrier function of the skin. To achieve this, defects need to be replaced by granulation tissue to form new connective tissue, and epithelial wound closure is required to restore the physical barrier. Different wound-healing phases are recognized, starting with an inflammation-dominated early phase giving way to granulation tissue build-up and scar remodeling after epithelial wound closure has been achieved. In the granulation tissue, mesenchymal cells are maximally activated, cells proliferate, and synthesize huge amounts of extracellular matrix. Epithelial cells also proliferate and migrate over the provisional matrix of the underlying granulation tissue, eventually closing the defect. This review focuses on the role of keratinocyte-fibroblast interactions in the wound-healing process. There is ample evidence that keratinocytes stimulate fibroblasts to synthesize growth factors, which in turn will stimulate keratinocyte proliferation in a double paracrine manner. Moreover, fibroblasts can acquire a myofibroblast phenotype under the control of keratinocytes. This depends on a finely tuned balance between a proinflammatory or a transforming growth factor (TGF)-beta-dominated environment. As the phenotype of fibroblasts from different tissues or body sites becomes better defined, we may understand their individual contribution in wound healing in more detail and possibly explain different clinical outcomes.
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            Tissue engineering for cutaneous wounds.

            Skin, the largest organ in the body, protects against toxins and microorganisms in the environment and serves to prevent dehydration of all non-aquatic animals. Immune surveillance, sensory detection, and self-healing are other critical functions of the skin. Loss of skin integrity because of injury or illness may result acutely in substantial physiologic imbalance and ultimately in significant disability or even death. It is estimated that, in 1992, there were 35.2 million cases of significant skin loss (US data) that required major therapeutic intervention. Of these, approximately 7 million wounds become chronic. Regardless of the specific advanced wound care product, the ideal goal would be to regenerate tissues such that both the structural and functional properties of the wounded tissue are restored to the levels before injury. The advent of tissue-engineered skin replacements revolutionized the therapeutic potential for recalcitrant wounds and for wounds that are not amenable to primary closure. This article will introduce the reader to the field of tissue engineering, briefly review tissue-engineered skin replacement from a historical perspective and then review current state-of-the-art concepts from our vantage point.
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              The role of myofibroblasts in wound healing.

              B Hinz (2016)
              The importance of proper skin wound healing becomes evident when our body's repair mechanisms fail, leading to either non-healing (chronic) wounds or excessive repair (fibrosis). Chronic wounds are a tremendous burden for patients and global healthcare systems and are on the rise due to their increasing incidence with age and diabetes. Curiously, these same risk factors also sign responsible for the development of hypertrophic scarring and organ fibrosis. Activated repair cells - myofibroblasts - are the main producers and organizers of extracellular matrix which is needed to restore tissue integrity after injury. Too many myofibroblasts working for too long cause tissue contractures that ultimately obstruct organ function. Insufficient myofibroblast activation and activities, in turn, prevents normal wound healing. This short review puts a spotlight on the myofibroblast for those who seek therapeutic targets in the context of dysregulated tissue repair. "Keep your myofibroblasts in balance" is the message.
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                Author and article information

                Journal
                Molecules
                Molecules
                molecules
                Molecules
                MDPI
                1420-3049
                22 April 2020
                April 2020
                : 25
                : 8
                : 1917
                Affiliations
                [1 ]Department of Biomedical Research, East-Slovak Institute of Cardiovascular Diseases, 040 11 Košice, Slovakia; ivankovac.kovi@ 123456gmail.com (I.K.); mcoma@ 123456vusch.sk (M.Č.); matomeister@ 123456gmail.com (M.H.); jan.durkac7@ 123456gmail.com (J.Ď.); lurban@ 123456vusch.sk (L.U.)
                [2 ]Second Department of Surgery, Louise Pasteur University Hospital and Pavol Jozef Šafárik University, 041 90 Košice, Slovakia
                [3 ]Laboratory of Cell Interactions, Center of Clinical and Preclinical Research, MediPark, Pavol Jozef Šafárik University, 040 11 Košice, Slovakia; nikola.melegova5@ 123456gmail.com (N.M.); mimagurbalova@ 123456gmail.com (M.G.)
                [4 ]Department of Pharmacology, Faculty of Medicine, Pavol Jozef Šafárik University, 040 11 Košice, Slovakia
                [5 ]Department of Human and Clinical Pharmacology, University of Veterinary Medicine and Pharmacy, 041 81 Košice, Slovakia; peter.takac@ 123456uvlf.sk
                [6 ]Department of Pathology, Louise Pasteur University Hospital, 041 90 Košice, Slovakia; katarina.kovacova@ 123456unlp.sk
                [7 ]Department of Chemical Theory of Drugs, Faculty of Pharmacy, Comenius University, 832 32 Bratislava, Slovakia; emilsva@ 123456yahoo.com
                [8 ]Department of Natural Drugs, Faculty of Pharmacy, University of Veterinary and Pharmaceutical Sciences, 612 42 Brno, Czech Republic
                [9 ]Department of Experimental Medicine, Faculty of Medicine, Pavol Jozef Šafárik University, 040 11 Košice, Slovakia; gabriela.mojzisova@ 123456upjs.sk
                [10 ]Prague Burn Centre, Third Faculty of Medicine and University Hospital Kralovske Vinohrady, Charles University, 100 34 Prague, Czech Republic; robert.zajicek@ 123456lf3.cuni.cz
                [11 ]Institute of Anatomy, First Faculty of Medicine, Charles University, 128 00 Prague, Czech Republic; szabopavol@ 123456gmail.com
                [12 ]BIOCEV-Biotechnology and Biomedical Centre of The Czech Academy of Sciences and Charles University, First Faculty of Medicine, Charles University, 252 50 Vestec, Czech Republic
                [13 ]Department of Pharmacognosy and Botany, Faculty of Pharmacy, Comenius University, 832 32 Bratislava, Slovakia
                Author notes
                [* ]Correspondence: mucaji@ 123456fpharm.uniba.sk (P.M.); pgal@ 123456vusch.sk (P.G.); Tel.: +421-25-011-7101 (P.M.); Fax: +421-55-789-1613 (P.G.)
                Author information
                https://orcid.org/0000-0003-4490-2615
                https://orcid.org/0000-0002-3398-3408
                Article
                molecules-25-01917
                10.3390/molecules25081917
                7221972
                32331226
                905c9c92-7b7d-4ddb-9815-3a3a7e37de5c
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 25 March 2020
                : 17 April 2020
                Categories
                Article

                wound healing,repair and regeneration,phytotherapy,horse chestnut

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