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      Metabolite profiling and wound-healing activity of Boerhavia diffusa leaf extracts using in vitro and in vivo models

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          Abstract

          Boerhavia diffusa is a perennial herb belonging to the Nyctaginaceae family. This plant has been widely used in Indian traditional medicinal system to cure several human ailments. However, traditional use of this plant in the treatment and management of wounds has not been validated by any comprehensive scientific study. The present study was aimed to explore the in vitro and in vivo wound healing potential of methanol extract (ME) and chloroform extract (CE) from B. diffusa leaf and subsequent identification of the bioactive metabolites, which might be responsible for enhancement of wound healing property of the extracts.

          The study included in vitro cell viability and wound scratch assays as well as in vivo excision wound assays in rat models. Both ME and CE were analysed for their antioxidant properties and phenolics content. The gas chromatography-mass spectrometry analyses were performed for identification of bioactive metabolites present in the ME and CE. ME of B. diffusa leaf significantly enhanced viability and migration of human keratinocyte cells (HaCaT) as compared to the untreated and CE-treated groups. The topical application of ME of B. diffusa leaf in excision wound model significantly decreased the wound area by the 14th day (91%) as compared to control (22%) ( p < 0.05). The GC-MS analysis revealed the presence of caffeic acid, ferulic acid and D-pinitol as the major bioactive metabolites in ME. These results suggest that ME of B. diffusa possesses significant wound healing potential, where D-pinitol and caffeic acid served as the lead constituent metabolites responsible for the healing.

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          Highlights

          • Reporting wound healing potential of Boerhavia diffusa leaf extracts.

          • GC-MS analyses of B. diffusa leaf extracts to identify bioactive wound healing metabolites.

          • D-Pinitol and caffeic acid were identified as lead metabolites triggering wound healing.

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          Most cited references40

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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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            Rat models of skin wound healing: a review.

            Rats have been widely used in the study of skin wound healing and the efficacy of different treatment modalities. This particular animal species is often selected for its availability, low cost, and small size. To define the current use of rat skin wound healing models, this manuscript provides a review of articles published between 2000 and 2003 that chose rats as their research animals. Of the 55 articles reviewed, it was found that 38.2% of the studies used incisional models and 38.2% used excisional models, with some studies using combinations. The majority of the studies (78.2%) used the rat's dorsum as the wound location. Male Sprague Dawley in the 250-300 gram weight range were the most preferred rats. Sodium pentobarbital/pentobarbitone was the most commonly used anesthetic choice. Similarities and differences in the selected experimental conditions are noted and questions are raised with regard to comparability between studies and the ability to transfer the data from the animal model to the human clinical situation. Attempts to compare studies for the advancement of wound healing knowledge are being hampered by the differences found between the studies. Standardization in reporting could facilitate comparisons and may instigate additional research that favors the inevitable comparisons between the studies. Thus, universal reporting requirements need to be developed for animal wound healing studies.
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              Comparative study on the antibacterial activity of phytochemical flavanones against methicillin-resistant Staphylococcus aureus.

              Differently substituted flavanones were isolated from Leguminosae and their antibacterial activity was comparatively studied against methicillin-resistant Staphylococcus aureus (MRSA). The minimum inhibitory concentrations (MICs) of phytochemical flavanones to clinical isolates of MRSA were determined by a serial agar dilution method. The structure-activity relationship has indicated that 2',4'- or 2',6'-dihydroxylation of the B ring and 5,7-dihydroxylation of the A ring in the flavanone structure are important for significant anti-MRSA activity and that substitution with a certain aliphatic group at the 6- or 8-position also enhances the activity. Among the thirteen flavanones tested, tetrahydroxyflavanones with these structural characteristics isolated from Sophora exigua and Echinosophora koreensis showed intensive activity to inhibit the growth of all MRSA strains at 3.13-6.25 micrograms/ml. The present hydroxyflavanones would be useful in the phytotherapeutic strategy against MRSA infections.
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                Author and article information

                Contributors
                Journal
                J Tradit Complement Med
                J Tradit Complement Med
                Journal of Traditional and Complementary Medicine
                Elsevier
                2225-4110
                19 February 2019
                January 2020
                19 February 2019
                : 10
                : 1
                : 52-59
                Affiliations
                [a ]Plant Molecular Biology Group, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India
                [b ]Molecular Endocrinology Laboratory, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India
                [c ]Department of Pharmacology, M. M. College of Pharmacy, Maharishi Markandeshwar (Deemed to be University), Mullana, Ambala, Haryana, 133203, India
                Author notes
                []Corresponding author. Plant Molecular Biology Group, Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand, 247667, India. debsrfbt@ 123456iitr.ac.in
                Article
                S2225-4110(18)30342-0
                10.1016/j.jtcme.2019.02.002
                6957803
                31956558
                35c497fd-d41b-4a23-990b-38e5ed38a8dd
                © 2019 Center for Food and Biomolecules, National Taiwan University. Production and hosting by Elsevier Taiwan LLC.

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 1 August 2018
                : 13 February 2019
                : 17 February 2019
                Categories
                Original Article

                topical ointment,hacat keratinocyte cell line,albino wistar rats,antioxidant,d-pinitol

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