Effect of medicinal plants on wound healing.

Effect of medicinal plants on wound healing. Arie Budovsky1, Ludmila Yarmolinsky1,2, Shimon Ben-Shabat2* 1

Judea Regional Research and Development Center, Carmel, 2Department of Biochemistry

and Pharmacology, Faculty of Health Sciences, Ben-Gurion University of the Negev, BeerSheva, Israel. Corresponding Author Shimon Ben-Shabat, Ph.D. Associate Professor, Department Of Biochemistry & Pharmacology, Faculty Of Health Sciences, Ben-Gurion University Of The Negev P.O. Box 653 Beer-Sheva 84105, Israel. Tel: 972-8-647-9354 ; 972-8-647-7363 Fax: 972-8-647-2984 E-Mail: [email protected]

This article has been accepted for publication and undergone full peer review but has not been through the copyediting, typesetting, pagination and proofreading process which may lead to differences between this version and the Version of Record. Please cite this article as an ‘Accepted Article’, doi: 10.1002/wrr.12274

This article is protected by copyright. All rights reserved.

Manuscript under review - CONFIDENTIAL

ABSTRACT In the United States alone chronic wounds affect 6.5 million patients. It is expected that the number of chronic wounds will increase worldwide due to the increase in age-related conditions and pathologies such as diabetes, obesity, and cardiovascular diseases. An estimated excess of US$25 billion is spent annually on treatment of chronic wounds, and the burden is rapidly growing due to increasing health care costs, an aging population, and a sharp rise in the incidence of diabetes and obesity worldwide. While current therapeutic agents have generally inadequate efficacy and number of serious adverse effects, the medicinal plants have been used in medicine since ancient times and are well known for their abilities to promote wound healing and prevent infection without grave side effects. Thus, herbal therapy may be an alternative strategy for treatment of wounds. The purpose of this review is to provide the verified data on the medicinal plants of the world flora with wound healing activity including the biologically active substances belonging to these herbal preparations, and describe in detail the various cellular and molecular mechanisms of their actions.

Keywords: Wound healing-plant extracts- phytochemicals- alkaloids- flavonoids-terpenesglycosides

Wound Repair and Regeneration This article is protected by copyright. All rights reserved.

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1. Introduction Out of estimated 250,000 flowering plant species in the world (1), 15% have been evaluated phytochemically and only 6% have been screened for biological activity (2). While a relatively small portion of all plants have been used as medicinal agents, their importance should not be undermined as almost 65% of the world’s population has incorporated them into their primary modality of health care (3). Remarkably, about one-third of all traditional herbal medicines are intended for treatment of wounds or skin disorders, compared to only 13% of modern drugs (4). Of note, the beneficial effects of plant extracts on skin wound healing (WH) has also gained support from several experimental studies (5-11). Given that in the United States alone chronic wounds affect 6.5 million patients, and that that the number of chronic wounds is likely to grow worldwide due to the increase in age-related conditions and pathologies such as diabetes, obesity, and cardiovascular diseases (12), any therapeutic intervention aimed at facilitating the WH processes should not be disregarded. Here, we present a systematic and comprehensive review of the literature on the pro-WH activity of herbal extracts and plant derived chemicals from all corners of the World.

2. Cellular and molecular mechanisms of wound healing process. WH is a complex biological process consisting of a synchronized chain of molecular events aimed at repairing the damaged tissue and restoring its protective barrier function (13). In general, the wound repair occurs in almost all tissues after exposure to any kind of destructive stimulus. This is particularly relevant for the skin – an organ that sustains insult and injury throughout life. Wounds are physical, chemical or thermal injuries which result in breaking of skin integrity.



In all organs of mammals, the normal response to injury occurs in three overlapping but distinct classical stages: inflammation, new tissue formation, and remodeling (14). Tissue damage leads to immediate activation of the coagulation cascade, inflammatory pathways and immune system in order to prevent blood fluid loss, to remove dead tissues and to neutralize invading pathogens (15). During and after formation of the platelet plug and deposition of fibrin matrix, neutrophils infiltrate the wound site. After 2–3 days, monocytes appear in the wound and differentiate into macrophages which coordinate the next phase of wound repair (16). The second stage of WH occurs 2–10 days after injury and is characterized by cellular proliferation and migration of different cell types, migration of keratinocytes over the injured dermis. In parallel to this migration angiogenesis starts as newly emerging capillaries replace the fibrin matrix and the granulation tissue. Angiogenesis is positively stimulated by various growth factors such as VEGFA and FGF2 (17). Along with the keratinocytes, fibroblasts migrate to the tissue repair zone and upon stimulation by macrophages turn into myofibroblasts (18). In turn, myofibroblasts bring the edges of the wound closer to one another and in cooperation with fibroblasts produce extracellular matrix creating the scar tissue (19). A key pleiotropic cytokine TGFβ1 secreted by fibroblasts stimulates proliferation of fibroblasts and their trans-differentiation to myofibroblasts (20). The final stage of WH (remodeling) begins 2–3 weeks after injury and may last for a year or even more. In the framework of the remodeling, the inflammatory response is downregulated as most of the cells involved in the previous stage of wound repair undergo apoptosis, and the dead cells are replaced by collagen and other extracellular-matrix proteins (21).

The complexity of the wound repair process on the molecular level stems from the fact that modulation of hundreds of genes (gene deletion, full or partial loss-of-function mutations or gene overexpression) in mice and other model organisms result in either delayed or


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accelerated WH (22). Among these genes are many growth factors, pro-inflammatory cytokines and chemokines. Expression of these pro-inflammatory mediators (e.g. IL6 and IL8) may stand behind scar formation and fibrotic processes in adults, while their absence in newborns may promote perfect scarless WH (reviewed in 23). These deviations from regular WH could lead to diverse age-related pathological conditions, from slow or ineffective tissue repair to fibroproliferative responses (“the so called never healing wounds”) (24, 25). While formation of scars and fibrotic responses are from complete tissue regeneration, the incomplete WH in adults may hold some evolutionary advantage as quick restoration of tissue homeostasis is vital for the protection of the organism from pathogen invasion (26). In any case, several growth promoting and survival pathways become activated during WH in adults. Signal cascade activated by TGFβ1 and mediated by the SMAD and Wnt dependent pathways enhances scarring (in particular, in case of the cutaneous WH), while the expression of TGFβ1 is transient in the skin of the neonates (reviewed in 23). In support of this notion are the studies on the TGFβ1 knock-out mouse which showed that these mice have defects in the proliferative phase of WH as at 10 days after incisional injury there was reduced granulation of the tissue and reduced collagen deposition (27). On the other hand, overexpression of TGFβ1 in keratinocytes led to upregulation in the type I collagen expression and increased deposition of the scar tissue (28). Of note, fibroblasts from both hypertrophic scars and keloids consistently overexpress proteins involved in TGFβ signal transduction (29). Thus, a prolonged and/or excessive secretion of TGFβ1 in the wound area may redirect the regular process of WH towards fibrosis (30, 31, 32). The hyper-activation of the Wnt pathway by TGFβ1 stands behind excessive scarring reminiscent of keloids and hypertrophic scars in transgenic mice that constitutively express the β-catenin (33). In contrast, animals with the conditional deletion of β-catenin, display much smaller wounds and have fewer fibroblasts in their granulation tissue (34). Modulation of the



TGFβ molecules can also bring beneficial effects. Neutralization of the TGFβ1 and TGFβ2 molecules by administration of antibodies to incisional rat wounds resulted in a significant reduction of extracellular matrix deposition and subsequent scarring (35). The same effect was achieved by applying recombinant TGFβ3 to the wound area (35). Thus, manipulations and interventions aimed at modifying the activity and level of WH/fibrosis associated proteins might lessen or even abrogate scar tissue generation along with creating conditions for successful WH.

Anti-platelet agents, aspirin or non-steroidal anti-inflamatory drugs are the most prescribed in the coagulation phase (36). Glucocorticoids are also used in many cases, they inhibit production of hypoxia-inducible factor-1 (HIF-1) (37), but they may promote wound infection (38) and have side effects on central nervous system (39), so administration of antibiotics and antiseptics is necessary. Some chemotherapeutic drugs are widespread in order to inhibit cellular metabolism, rapid cell division, and angiogenesis, but they significantly decrease functions of the immune system, and often cause excessive bleeding at the wound site (38). Current treatments for delayed WH (old age, diabetes or radiation exposure), excessive healing (hypertrophic and keloid scars) and fibroproliferative diseases include neutralization of pro-inflammatory mediators (25), silicone and pressure dressings (40), vitamin E supplementation (41), administration of corticosteroids (42), 5-fluorouracil (43), bleomycin (44), and others, reviewed by (45). While meeting some success, these treatments and pharmacological interventions still have various adverse effects. In fact, single-agent therapies, such as administration of a growth factor, have only a moderate impact on WH, most probably because of the considerable complexity of the wound repair process and redundancy in associated components and pathways. Since no “magic bullet” has been found so far to treat abnormal WH and related processes, we should not neglect herbal


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agents that might on one hand accelerate wound closures and on the other one might reduce fibrosis. From this point of view, the application of whole, fractionated plant extracts, or even selected compounds of herbal origin could be highly advantageous as herbaceuticals may have multiple targets, thus indicating the possibility of pleiotropic beneficial effects on WH (4).

3. Medicinal plants of the world flora with wound healing activity. Medicinal plants have been widely acknowledged in medicine since ancient times, at least for 60 000 years (46). Many ancient civilizations used plants for stimulation of WH. For example, ancient Egyptians used Aloe vera Mill in treating wounds as early as 1500 B.C (47). Many experimental and clinical studies demonstrated WH properties of the Aloe vera Mill gel and latex which were obtained from a mucilaginous tissue of its leaves (48). The possible mechanisms by which the gel helped wound healing included keeping the wound moist, increased epithelial cell migration, more rapid maturation of collagen, and reduction in inflammation (49); increased collagen synthesis (50, 51); increase in synthesis of hyaluronic acid and dermatan sulfate in the granulation tissue of the healing wound (50) and increased blood supply as a result of improved oxygenation (52). Mannose rich polysaccharides of Aloe vera Mill gel enhanced TIMP-2 gene expression, collagen, NAGLA and NAGA synthesis during the skin wound repair of rats (53). Although Aloe vera Mill is an outstanding example of well-studied wound-healing plant, many of its active phytochemicals have not been identified yet. Leaves extracts of other species of Aloe such as Aloe literalis Baker (54), Aloe arborescens Mill, Aloe excelsa Berger and Aloe ferox Mill have also significant WH properties (55).



The WH activity of 13 Verbascum species was assessed in vivo using two models (linear incision and circular excision); V.olympicum Boiss, V. stachydifolium Lam and V. uschackense Lam had the highest activities on the both wound models and the methanolic extracts of V. latisepalum Boiss, V. mucronatum Lam and V. pterocalycinum var. mutense demonstrated promising pro-WH potential (10). Although mechanisms of pro-WH actions of the Verbascum species remain unknown, some active phytochemicals were determined in the case of V. mucronatum Lam (56).

While it is known that various combinations of extracts may be highly beneficial, few researches have been devoted to studying the synergistic interactions between the active extracts and other phytochemicals. Nevertheless, it was shown that ethanol based mixed extract of Curcuma longa L., Areca catechu L., Oryza sativa L., and Garcinia mangostana L. had very low toxicity along with good pro-WH properties in vitro (57). Extract from the mixture of Radix astragali Schishkin and Radix Rechmanniae Libosch in the 2:1 ratio promoted keratinocyte proliferation by regulating expression of growth factor receptors (58). Another factor regulating the WH potential of botanicals is the exposure of plants to the stressful conditions. It is known that plants under stress are able to produce more bioactive compounds. For example, Phlomis viscosa Poiret grown in stressful phytogeographic zone in Judea region (Israel) demonstrated high pro-WH activity (59). Altogether, in the framework of this manuscript, we have collected data on different kinds of wound-healing promoting plant extracts (aqueous, ethanolic, methanolic and so on) from various plant organs (roots, stems, leaves, buds, flowers, fruits, and so on). The major findings related to WH activities of herbal extracts are summarized in Table 2. Only crude extracts were included in Table 1 as the active compounds which are responsible for the WH properties of these plants have not been isolated, purified and identified yet.


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4. Plant phytochemicals with wound healing activity A wide variety of bioactive constituents of different structures stands behind the therapeutic activity of the medicinal plants: polyphenols (flavonoids, phenolic acids, lignans, tannins, stilbenes and coumarins);

terpenes, sulfur-containing compounds (sulfides and

glucosinolates), carotenoids, saponins, furils, alkaloids, polyines, thiophenes, different sugars, fatty oils, resins, phytosterols, proteins, peptides and etc (60). In particular, review of Ghosh and Gaba (61) analyzes WH properties of several plant compounds including flavonoids, quinones, phenolic acids, phenyl propanoids, terpenoids, tannins and sugars demonstrating that the main effects of these active compounds of plants were connected to their antimicrobial activity, antioxidant properties and their abilities to enhance cell proliferation, collagen production and DNA synthesis. It is reasonable to expect that the majority of plant phytochemicals have a great wound-healing potential, however in many cases their active molecules have not been identified yet, and little is known about their modes of action. Among plant phytochemicals, WH promoting activity of alkaloids, flavonoids, terpenes and glycosides is better researched than that of other bioactive plant constituents. Therefore, we present data on the more studied phytochemicals.

4.1. Alkaloids Alkaloids are nitrogen-containing phytochemicals isolated and identified from plants. Some alkaloids have an important role in the process of WH. Many alkaloids are considered as very toxic phytochemicals. For example, nicotine is found in the nightshade family of plants (Solanaceae). Nevertheless, it can improve WH and accelerate angiogenesis in small doses (62) due to stimulating many intracellular processes. Specifically, nicotine binds to the nAChR and increases intracellular level of calcium (63).



The effects of the Aconitum baikalense Turcz alkaloids (mesaconitine, hypaconitine, songorine, napelline, and 12-epinapelline N-oxide) (Figure 1) on functional activity of fibroblast precursors were studied in vitro (64). Exposure to some of these alkaloids (songorine, napelline and hypaconitine) led to direct stimulation of the fibroblast precursors and to higher production of growth factors by the skin stromal cells (65). Another alkaloid taspine (Figure 2) found in various plants including Magnolia, enhanced WH by increasing the autocrine activity of TGF-beta1 and EGF on fibroblasts (66).

4.2. Flavonoids Flavonoids are oxygen-containing aromatic compounds which WH properties are wellknown. These compounds are able to promote the rapid WH due to their antimicrobial, antioxidant and astringent properties (67). Quercetin, found in variety of plants, stimulated incorporation of collagen matrices in case of dermal WH (68). Mixture of flavonoids (quercetin, isorhamnetin and kaempferol), obtained from Hippophae rhamnoides L, improved collagen deposition and maturation, and decreased COX-2 level (69). Later, the WH properties of this plant were also attributed to the gallic acid which stimulated the increase in ascorbic acid contents which in turn is important for collagen metabolism (70). Additional quercetin based flavonoid mixture (quercetin-3-O-[(6-caffeoyl)-β-glucopyranosyl (1 → 3) αrhamnopyranoside]-7-O-α-rhamnopyranoside,

kaempferol-3-O-[(6-caffeoyl)-β-

glucopyranosyl (1 → 3) α-rhamnopyranoside]-7-O-α-rhamnopyranoside and quercetin-3-Omethyl) isolated from the aerial parts of the fern Ophioglossum vulgatum L. proved to be active in a scratch-wound healing assay on keratinocytes through intracellular calciumdependent, ERK1/2 MAP kinase dependent mechanism (71). Furthermore, quercetin 3-Oglucoside isolated from the leaves of Sambucus ebulus L was determined as one of the active components of the WH as determined by animal experiments. Specifically, this agent enhanced re-epithelization (9).


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Another flavone - 2-(3,4-dihydroxy-phenyl-5; 7—dihydroxy-chromen-4 is an active constituent of the Indian pro-WH plant, Pedilanthus tithymaloides L. It promotes WH by accelerating epithelization, scavenging of free radicals, and inhibiting several mediators of inflammatory pathways including the NF-κB activity (72). It was also found that Vicenin-2 flavonoid (isolated from crude methanol extract of food plant Moringa olifera,) was able to enhance the proliferation and viability of human fibroblast cells in vitro which led to subsequent faster WH (73).

Various Hypericum species plants are acknowledged as popular folk remedies for stimulation of WH. Examination of Hypericum scabrum L and Hypericum perforatum L oil extracts for the WH activity showed that the latter plant and its flavonoids (hyperoside, isoquercitrinrutin and epicatechin) indeed are pro-WH (8).

Another popular folk remedy is Silybum marianum L, as investigations into silibinin from Silybum marianum revealed its ability to promote a faster WH process by regulation of STM1 gene (encodes stromelysin-1 which belongs to the metalloproteinase family and has an important role in WH) expression, acetyl hexoseamonine and collagen production (74).

While in case of various flavonoids the mechanism of action is still obscure, studies of 4’,6,7trimethoxyisoflavone (Figure 3), isolated from the Euchresta formosana Ohwi, showed that this flavonoid increased rate of WH by promoting migration, but not proliferation of keratinocytes through induction of NADPH oxidases (NOXs) (the expression both at the mRNA and protein levels was examined) (75).



4.3. Terpenes Terpenes are large classes of organic compounds, produced by a variety of plants, which to a great extent exist in form of essential oils. Gentiana lutea L contains several various terpenes including gentiopicroside, sweroside and swertiamarine which synergistically promote WH stimulating collagen production and the mitotic activity of embryonic fibroblasts (Figure 4) (76). Many other monoterpenes also have beneficial effects on skin WH including ajugol (56), borneol (77), catalpol (56, 78), thymol (79), genipin (80), lasianthoside (56), alphabisabolol (80), alpha-terpiniol (81), and aucubin (56, 82), but the precise mechanisms of their actions are mostly not known. Pro-WH properties of borneol might be associated with its ability to suppress the proinflammatory cytokines at the mRNA level (83). Thymol pro-WH activity could be explained by modulatory effect on the fibroblast metabolism and collagen synthesis (79).

With regard to the more complex terpenes, two lupane triterpenoids from Paullinia pinnata L. 6beta-(3'-methoxy-4'-hydroxybenzoyl)-lup-20(29)-ene-one, and 6beta-(3'-methoxy-4'hydroxybenzoyl)-lup-20(29)-ene-ol had a significant fibroblast stimulatory activity (84). The triterpene oleanolic acid (isolated from Viscum album L) influenced migratory activity of NIH/3T3 Fibroblasts and Hecate Keratinocytes. This in turn accelerated wound closure. Nevertheless, mechanisms behind the beneficial effects of these glycosides remain to be investigated (85). The triterpeniod compounds (asiatic acid, asiaticoside, madecassic acid and madecassoside) isolated from Centella asiatica L promoted WH through induction of expression of genes involved in angiogenesis and the remodeling of extracellular matrix, as well as diverse growth factors (86). In particular, madecassoside’s and asiaticoside’s pro-WH activities could be associated with increased collagen synthesis mediated via activation of the TGF-β/Smad signaling pathway (87, 88). Other known monoterpene derivatives showing


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beneficial effects on intestinal WH are the cannabinoids from Cannabis sativa L. It was found that tetrahydrocannabinoid (THC) enhanced epithelial wound closure (89). Cannabidiol (CBD) the non-psychoactive compound in Cannabis sativa L is also well known for its beneficial effects on intestinal inflammation as well as reducing cytokine release and promoting WH (90,91).

4.4. Glycosides Glycosides are conjugates of sugars with small organic molecules which can be classified by the glycone groups, types of glycosidic bonds, and by the aglycone groups. A classification by the aglycone group is the most useful one as there are several approaches to this classification (92, 93). Many WH associated glycosides have already been mentioned in previous chapters, in particular the flavonoid glycosides.

Many plants are renowned for their pro-WH glycosides. For example, the mixture of the galactoglycerolipids from the fern Ophioglossum vulgatum L. (dominant compound 1,2-di-Olinolencyl-3-O-B-D-galacto-pyranosyl-glycerol) had pro-WH activity and mechanism of their action was similar to that of the flavonoid glycosides (94). Mixture of 11 saponin glycosides from Panax ginseng promoted re-epitalization of the skin wounds in animal models, enhanced collagen synthesis in skin fibroblasts through phosphorylation of Smad 2 protein, and inhibited inflammatory processes at the early phases of WH (95). Compounds from genetically modified flax seeds with high glycoside content (secoisolariciresinol diglycoside, p-coumaric acid glycosides, ferulic acid glycoside, and caffeic acid glucosides) enhanced proliferation and migration of normal human dermal fibroblasts as determined by the wound scratch assay (96). Two steroidal glycosides from Lilium longiflorum Thunmb ((22R,25R)spirosol-5-en-3β-yl

O-α-L-rhamnopyranosyl-(1-2)-β-D-glucopyranosyl-(1-4)-β-D-



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glucopyranoside, and (22R,25R)-spirosol-5-en-3β-yl-O-α-L-rhamnopyranosyl-(1-2)-[6-Oacetyl-β-D-glucopyranosyl-(1-4)]-β-D-glucopyranoside) which have very similar structures (figure 5) could influence the fibroblast migration, NO production and TGF-β receptor I mRNA expression (97). It is interesting to note that the analysis of these compounds in different organs of L. longiflorum demonstrated their highest concentrations in flower buds (acknowledged in folk medicine as WH-remedy) and lower stems (98).

Phenylpropanoid glycosides are widespread phytochemicals of many plants from Asteraceae, Labiateae, Liliceae, Oleaceae families. For example, Verbascum mucronatum is rich in various glycosides including pro-WH one called verbascoside (56). It was shown that this phenylpropanoid glycoside is able to enhance hepatocyte growth factor production in human dermal fibroblasts (99). The WH properties of this compound are connected to its chemical nature ensuring stronger affinity for negatively charged membranes composed of phosphatidylglycerol (100). Verbascoside was also shown to possess pro-WH properties in conjunction

with

another

phenylpropanoid

glycoside

teupolioside

(produced

biotechnologically from Syringa vulgaris L and Ajuga reptans L plant cell cultures) (101) and with luteolin-7-O-glucoside (found in methanol extract of Buddleja globosa Hope) (102).

5. Human targets of herbal pro-WH compounds It is reasonable to suggest that the biological effects of the plant chemicals are to a great extent associated with their protein targets. With this in mind, we have first extracted the data on reported pro-WH compounds available in scientific literature, and then determined the proteins interacting with these chemicals, using the STITCH database - . http://stitch.embl.de/ (103-105) which contains a consolidated set of chemicals assigned with PubChem ID number along with their interactions with proteins (physical interactions).


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Out of more than 30 pro-WH compounds discussed in this work, almost half targeted genes associated with WH modulation (106, 107). Remarkably, 110 out of 207 of the above mentioned WH associated genes were targeted in a pleotropic and synergistic manner by the pro-WH herbal compounds including flavonoids such as curcumin, epicathechin gallate, resveratrol, and quercitin among others. The selected compounds and their respective targets were organized in Table 2 which also includes short description of the targets’ relevance to the WH process. Thus, the pro-WH effects of the described compounds are mediated by their direct involvement in the core WH processes. Subsequently, these interactions and their impact on human proteome/interactome should be a point of future investigations.

6. Clinical trials Although the benefit of medicinal plants and their compounds is confirmed by various experimental studies, few clinical trials are known. In contrast to 62 experimental studies, only 3 human clinical studies were mentioned in literature (108). These studies focused on the effect of medicinal plants on WH of burns as they investigated effect of Aloe vera gel. However, the authors could not reach definite conclusions due to the small number of patients and low methodological level. Nevertheless, the positive effects of dressing with Aloe vera gel on cesarean WH was shown in prospective randomized double-blind clinical trial (109). Another clinical study showed that using Aloe vera and Calendula ointment significantly increased the speed of episiotomy WH (110). Successful clinical trials are summarized in Table 3.

Conclusions This review demonstrates that a wide plethora of medicinal plants from all over the World and the phytochemicals that they contain (alkaloids, flavonoids, terpenes, glycosides) have



beneficial pro-WH properties both in in vivo and in vitro various models. Thus, they seem to be promising candidates for developing new WH drugs targeting the WH associated human proteins. In case of other phytochemicals, it may be too early to reach such a conclusion since their purification and identification have not yet been completed, and their modes of actions were not investigated. For example, aqueous extract from Ananas comosus L (pineapple) crown leaves demonstrated beneficial pro-WH properties which were associated with activity of several proteins which structure and modes of action still remain unknown (111). It is important to stress that the obvious weakness of current state of research is incomplete understanding of mechanisms behind pro-WH activity of many herbal compounds. Based on the above-mentioned studies, we conclude that the majority of the investigated plant extracts contain at least several compounds responsible for their pro-WH activity, and thus their synergistic and pleiotropic activity should be the focus of future investigations.

Abbreviations B.C Before Christ FGF2 Basic fibroblast growth factor EGF Epidermial growth factor FGF2 Fibroblast growth factor 2 HIF-1 Hypoxia-inducible factor IL6


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Interleukin 6 IL8 Interleukin 8 NADPH oxidase Nicotinamide adenine dinucleotide phosphate-oxidase NAGA n-acetyl glucosamine NAGLA n-acetyl galactosamine NF- kB nuclear factor kappa-light-chain-enhancer of activated B cells NOX NADPH oxidase protein family ROS Reactive oxygen species SMAD Intracellular proteins that transduce extracellular signals from transforming growth factor beta ligands to the nucleus STM1 Stromelysin-1 gene TGFβ1 Transforming growth factor beta TIMP-2 metallopeptidase inhibitor 2 VEGFA



Vascular endothelial growth factor A WH Wound healing Wnt Group of signal transduction pathway


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References

1. Cronquist A. An Integrated System of Classification of Flowering Plants. New York: Columbia University Press, 1981. 2. Verpoorte R. Pharmacognosy in the new millennium: lead finding and biotechnology. J Pharm Pharmacol 2000; 52: 253-62. 3. Farnsworth NR, Akerele O, Bingel AS, Soejarto DD, Guo Z. Medicinal plants in therapy. Bull World Health Organ. 1985; 63: 965-81. 4. Mantle D, Gok MA, Lennard TW. Adverse and beneficial effects of plant extracts on skin and skin disorders. Adverse Drug React Toxicol Rev 2001;20(2 Suppl):89-103. 5. Yeşilada E, Honda G, Sezik E, Tabata M, Goto K, Ikeshiro Y.Traditional medicine in Turkey. IV. Folk medicine in the Mediterranean subdivision. J Ethnopharmacol 1993; 39: 31-8. 6. Fronza M, Heinzmann B, Hamburger M, Laufer S, Merfort I. Determination of the wound healing effect of Calendula extracts using the scratch assay with 3T3 fibroblasts. J Ethnopharmacol 2009; 126: 463-7.

7. Linde K. St. John's wort - an overview. Forsch Komplementmed. 2009; 16: 146-55. 8. Suntar IP, Akkol EK, Yilmazer D, Baykal T, Kirmizibekmez H, Alper M, Yeşilada E. Investigations on the in vivo wound healing potential of Hypericum perforatum L. J Ethnopharmacol. 2010; 127: 468-77. 9. Suntar IP,Akkol EK, Yalçin FN, Koca U, Keleş H, Yesilada E. Wound healing potential of Sambucus ebulus L. leaves and isolation of an active component, quercetin 3-O-glucoside J Ethnopharmacol 2010;129:106-14. 10. Suntar I, Tatli II, Kupeli Akkoy E, Keleş H, Kahraman C, Akdemir Z. An ethnopharmacological study on Verbascum species from conventional wound healing use to scientific verification. J Ethnopharmacol. 2010; 132: 408-13.



11. Suntar I, Koca U, Keleş H, Akkol EK. Wound Healing Activity of Rubus sanctus Schreber (Rosaceae): Preclinical Study in Animal Models. Evid Based Complement Alternat Med. 2011; 816156: 1-10. 12. Sen CK, Gordillo GM, Roy S, Kirsner R, Lambert L, Hunt TK, Gottrup F, Gurtner GC, Longaker MT. Human skin wounds: a major and snowballing threat to public health and the economy. Wound Repair Regen.2009; 17(6 Suppl):763-71 13. Gurtner GC, Werner S, Barrandon Y, Longaker MT. Wound repair and regeneration. Nature 2008; 453(7193 Suppl.):314-21. 14. Diegelmann RF, Evans MC. Wound healing: an overview of acute, fibrotic and delayed healing. Front Biosci 2004; 1:283-9. 15. Martin P, Leibovich SJ. Inflammatory cells during wound repair: the good, the bad and the ugly. Trends Cell Biol 2005; 15(11 Suppl):599-607. 16. Cohen HB, Mosser DM.Extrinsic and intrinsic control of macrophage inflammatory responses. J Leukoc Biol 2013;.;94(5 Suppl):913-9. 17. Werner S, Grose R. Regulation of wound healing by growth factors and cytokines. Physiol Rev 2003; 83(3 Suppl):835-70. 18. Opalenik SR, Davidson JM. Fibroblast differentiation of bone marrow-derived cells during wound repair. FASEB J 2005; 11:1561-3. 19. Werner S, Krieg T, Smola H. Keratinocyte-fibroblast interactions in wound healing. J Invest Dermatol 2007; 127(5 Suppl):998-1008. 20. Werner S, Grose R. Regulation of wound healing by growth factors and cytokines. Physiol Rev 2003; 83(3 Suppl):835-70. 21. Agha R, Ogawa R, Pietramaggiori G, Orgill DP. A review of the role of mechanical forces in cutaneous wound healing. J Surg Res 2011; 171(2 Suppl):700-8.


Page 20 of 47

Page 21 of 47


22. Yanai H, Budovsky A, Tacutu R, Fraifeld VE. Is rate of skin wound healing associated with aging or longevity phenotype? Biogerontology 2011; 12(6 Suppl):591-7. 23. Profyris C, Tziotzios C, Do Vale I. Cutaneous scarring: Pathophysiology, molecular mechanisms, and scar reduction therapeutics Part I. The molecular basis of scar formation. J Am Acad Dermatol2012.; 66(1 Suppl):1-10. 24. Diegelmann RF, Evans MC. Wound healing: an overview of acute, fibrotic and delayed healing. Front Biosci. 2004; 1(9 Suppl): 283-9. 25. Ferguson MW, O'Kane S. Scar-free healing: from embryonic mechanisms to adult therapeutic intervention. Philos Trans R Soc Lond B Biol Sci 2004; 359(1445 Suppl):839-50. 26. Baker DJ, Jeganathan KB, Cameron JD, Thompson M, Juneja S, Kopecka A, Kumar R, Jenkins RB, de Groen PC, Roche P, van Deursen JM.BubR1 insufficiency causes early onset of aging-associated phenotypes and infertility in mice. Nat Genet 2004; 36(7 Suppl):744-9. 27. Brown RL, Ormsby I, Doetschman TC, Greenhalgh DG. Wound healing in the transforming growth factor-beta-deficient mouse. Wound Repair Regen 1995; 3(1 Suppl):25-36. 28. Yang L, Chan T, Demare J, Iwashina T, Ghahary A, Scott PG, Tredget EE. Healing of burn wounds in transgenic mice overexpressing transforming growth factor-beta 1 in the epidermis. Am J Pathol 2001; 159(6 Suppl):2147-57. 29. Chalmers RL. The evidence for the role of transforming growth factor-beta in the formation of abnormal scarring. Int Wound J 2011; 8(3 Suppl):218-23. 30. Beanes SR, Dang C, Soo C, Ting K. Skin repair and scar formation: the central role of TGF-beta. Expert Rev Mol Med 2003; 5:1-22.



31. Wynn TA . Common and unique mechanisms regulate fibrosis in various fibroproliferative diseases. J Clin Invest 2007; 117: 524-9. 32. Metcalfe AD, Ferguson MW. Tissue engineering of replacement skin: the crossroads of biomaterials, wound healing, embryonic development, stem cells and regeneration. J R Soc Interface 2007; 4: 413-37. 33. Cheon SS, Cheah AY, Turley S, Nadesan P, Poon R, Clevers H, Alman BA. betaCatenin stabilization dysregulates mesenchymal cell proliferation, motility, and invasiveness and causes aggressive fibromatosis and hyperplastic cutaneous wounds. Proc Natl Acad Sci U S A 2002;99(Suppl10):6973-8. 34. Cheon SS, Wei Q, Gurung A, Youn A, Bright T, Poon R, Whetstone H, Guha A, Alman BA. Beta-catenin regulates wound size and mediates the effect of TGF-beta in cutaneous healing. FASEB J 2006; 20(6 Suppl):692-701. 35. Shah M, Foreman DM, Ferguson MW. Neutralisation of TGF-beta 1 and TGF-beta 2 or exogenous addition of TGF-beta 3 to cutaneous rat wounds reduces scarring. J Cell Sci 1995; 108 (3 Suppl):985-1002. 36. Giannobile WV, Somerman MJ . Growth and amelogenin-like factors in periodontal wound healing. A systematic review. Ann. Periodontol 2003; 8(1 Suppl): 193-204. 37. Wagner AE, Huck G, Stiehi DP, Jelkmann W, Hellwig-Burkel T. Dexamethasone impairs hypoxia-inducible factor-1 function. Biochem. Biophys. Res. Commun 2008 372 (2 Suppl): 336-40. 38. Guo S, Dipietro LA . Factors affecting wound healing. J. Dent. Res 2010; 89 (3Suppl): 219-29. 39. Ciriaco M, Ventrice P, Russo G, Scicchitano M, Mazzitello G, Scicchitano F, Russo E. Ventrice P. Corticosteroid-related central nervous system side effects. J. Pharmacol Pharmacotherap;2013; 4 (1 Suppl): 494-8.


Page 22 of 47

Page 23 of 47


40. O'Brien L, Pandit A. Silicon gel sheeting for preventing and treating hypertrophic and keloid scars. Cochrane Database Syst Rev 2006;1:CD003826. 41. Baumann LS, Spencer J. The effects of topical vitamin E on the cosmetic appearance of scars. Dermatol Surg 1999; 25(4 Suppl):311-5. 42. Widgerow AD, Chait LA, Stals R, Stals PJ. New innovations in scar management or Burn Wound Healing of Centella asiatica Herbs. Aesthetic Plast Surg 2000;24(3 Suppl):227-34. 43. Kontochristopoulos G, Stefanaki C, Panagiotopoulos A, Stefanaki K, Argyrakos T, Petridis A, Katsambas A. Intralesional 5-fluorouracil in the treatment of keloids: an open clinical and histopathologic study. J Am Acad Dermatol 2005; 52(3 Suppl):474-9. 44. Cutroneo KR, White SL, Phan SH, Ehrlich HP. Therapies for bleomycin induced lung fibrosis through regulation of TGF-beta1 induced collagen gene expression. J Cell Physiol 2007; 211(3 Suppl):585-9. 45. Tziotzios C, Profyris C, Sterling J. Cutaneous scarring: Pathophysiology, molecular mechanisms, and scar reduction therapeutics Part II. Strategies to reduce scar formation after dermatologic procedures. J Am Acad Dermatol 2012;66(1 Suppl):13-24. 46. Fabricant DS, Farnsworth NR. The value of plants used in traditional medicine for drug discovery. Environ. Health Perspect 2001; 109 (1 Suppl): 69-75. 47. Sharma Y, Jeyabalan J, Singh R.Potential wound healing agents from medicinal plants: a review. Pharmacologia 2013; 4 (5): 1-10. 48. Gupta VK, Malhorta S . Pharmacological attribute of Aloe vera: Revalidation through experimental and clinical studies. Ayu 2012; 33 (2 Suppl): 193-6. 49. Reynolds T, Dweck AC. Aloe vera leaf gel: a review update. J Ethnopharmacol. 1999;68:3-37.



50. Chithra P, Sajithalal GB, Ghandrakasan G. Influence Aloe Vera, on collagen turnover in healing of dermal wounds in rats. Indian J. Exp. Biol 1998; 36 (3 Suppl): 896-901. 51. Chithra P, Sajithalal GB, Ghandrakasan G. Influence Aloe Vera, on collagen turnover in healing of dermal wounds in rats. Influence Mol. Cell Biochem 1998;181: 71-6. 52. Davis RH, Leitner MG, Russo JM, Byrne ME. Wound healing. Oral and topical activity of Aloe vera. J Am Podiatr Med Assoc 1989; 79(11 Suppl):559-62. 53. Tabandeh MR, Oryan A, Mohammadalipour A. Polysaccharides of Aloe vera induce MMP-3 and TIMP-2 gene expression during the skin wound repair of rat. Int J Biol Macromol 2014;65:424-30. 54. Hajhashemi V, Ghannadi A, Heidari AH. Anti-inflammatory and wound healing activities of Aloe littoralis in rats. Res Pharm Sci 2012; 7(2 Suppl): 73-8. 55. Coopoosamy RM, Naidoo KK . A comparative study of three Aloe species used to treat skin diseases in South African rural communities. J Altern Complement Med 2013; 19(5 Suppl):425-8. 56. Akdemir Z, Kahraman C, Tatlı II, Küpeli Akkol E, Süntar I, Keles H. Bioassay-guided isolation of anti-inflammatory, antinociceptive and wound healer glycosides from the flowers of Verbascum mucronatum Lam J Ethnopharmacol 2011; 136(3Suppl):436-43. 57. Chushri S, Settharaksa S, Chokpaisarn J, Limsuwan S, Voravuthikunchai SP. Thai herbal formulas used for wound treatment: a study of their antibacterial potency, antiinflammatory, antioxidant, and cytotoxicity effects. J Altern Complement Med 2013; 19(7 Suppl):671-6. 58. Ren JW, Chan KM, Lai PK, Lau CB, Yu H, Leung PC, Fung KP, Yu WF, Cho CH. Extracts from Radix Astragali and Radix Rehmanniae promote keratinocyte proliferation by regulating expression of growth factor receptors Phytother Res 2012; 26(10 Suppl):1547-54.


Page 24 of 47

Page 25 of 47


59. Budovsky A, Shteinberg A, Maor H, Duman O, Yanai H, Wolfson M, Fraifeld V , Shteinberg A, Maor H, Duman O, Yanai H, Wolfson M, Fraifeld V. Uncovering the Geroprotective

Potential

of

Medicinal

Plants

from

the

Judea

Region

(Israel).Rejuvenation Res. 2014; 17(2 Suppl):134-9. 60. Rostugno MA, Prado JM. Natural product extraction: principles and applications. RSC Publishing, 2013. 61. Ghosh PK, Gaba A. Phyto-extracts in wound healing. J Pharm Pharm Sci. 2013;16(5 Suppl):760-820. 62. Morimoto N, Takemoto S, Kawazoe T, Suzuki S. Nicotine at a low concentration promotes wound healing. J Surg Res 2008;145(2 Suppl):199-04. 63. Tournie JM, Maouche K, Coraux C, Zahm JM, Cloëz-Tayarani I, Nawrocki-Raby B, Bonnomet A, Burlet H, Lebargy F, Polette M, Birembaut P.

Alpha3alpha5beta2-

Nicotinic acetylcholine receptor contributes to the wound repair of the respiratory epithelium by modulating intracellular calcium in migrating cells. Am J Pathol 2006 ;168(1 Suppl):55-68. 64. Nesterova YV, Povetieva TN, Suslov NI, Zhdanov VV, Hrichkova TY, Udut EV, Chaykovskiy AS, Gaydamovich NN, Andreeva TI, Dygai AM. Regeneratory characteristics of complex extract and isolated diterpene alkaloids of Aconitum baikalense. Bull Exp Biol Med 2012; 152(4 Suppl):439-43. 65. Zyuzkov GN, Krapivin AV, Nesterova YV, Povetieva TN, Zhdanov VV, Suslov NI, Fomina TI, Udut EV, Miroshnichenko LA, Simanina EV, Semenov AA, Kravtsova SS, Dygai AM . Mechanisms of regeneratory effects of baikal aconite diterpene alkaloids.Bull.Exp. Bio. Med 2012; 153 (8 Suppl): 846-50. 66. Dong Y, He L, Chen F. Enhancement of wound healing by taspine and its effect on fibroblast . Zhong Yao Cai 2005; 28(7 Suppl.):579-82.



Page 26 of 47

67. Tsuchiya H, Sato M., Miyazaki T, Fujiwara S, Tanigaki S, Ohyama M., Tanaka T, Iinuma, M. flavanones

Comparative study on the antibacterial activity of phytochemical against

methicillinresistant

Staphylococcus

aureus.

Journal

of

Ethnopharmacology 1996; 50: 27-34. 68. Gomathi K, Gopinath D, Rafiuddin Ahmed M, Jayakumar R. Quercetin incorporated collagen matrices for dermal wound healing processes in rat. Biomaterials 2003;24(16 Suppl):2767-72. 69. Fu SC, Hui CW, Li LC, Cheuk YC, Qin L, Gao J, Chan KM.( 2005). Total flavones of Hippophae rhamnoides promotes early restoration of ultimate stress of healing patellar tendon in a rat model. Med Eng Phys 2005; 27(4 Suppl):313-21. 70. Gupta A, Kumar R, Pal K, Singh V, Banerjee PK, Sawhney RC. Influence of sea buckthorn (Hippophae rhamnoides L.) flavone on dermal wound healing in rats. Mol Cell Biochem 2006; 290:193-8. 71. Clericuzio M, Tinello S, Burlando B, Ranzato E, Martinotti S, Cornara L, La Rocca A. Flavonoid oligoglycosides from Ophioglossum vulgatum L. having wound healing properties. Planta Med 2012;78(15 Suppl):1639-44. 72. Ghosh S, Samanta A, Mandal NB, Bannerjee S, Chattopadhyay D. Evaluation of the wound healing activity of methanol extract of Pedilanthus tithymaloides (L.) Poit leaf and its isolated active constituents in topical formulation J Ethnopharmacol 2012;142(3Suppl):714-22. 73. Muhammad AA, Pauzi NA, Arulselvan P, Abas F, Fakurazi S . In vitro wound healing potential and identification of bioactive compounds from Moringa oleifera Lam. Biomed Res Int 2013; 974580: 1-8. 74. Tabandeh MR, Oryan A, Mohhammad-Alipour A, Tabatabaei-Naieni A. Silibinin regulates matrix metalloproteinase 3 (stromelysine1) gene expression, hexoseamines


Page 27 of 47


and collagen production during rat skin wound healing. Phytother Res 2013; 27(8 Suppl):1149-53. 75. Bui NT, Ho MT, Kim YM, Lim Y, Cho M. Flavonoids promoting HaCaT migration: II. Molecular mechanism of 4',6,7-trimethoxyisoflavone via NOX2 activation. Phytomedicine 2013; 13: 419-23. 76. Ozturk N, Korkmaz S, Oztürk Y, Başer KH. Effects of gentiopicroside, sweroside and swertiamarine, secoiridoids from gentian (Gentiana lutea ssp. symphyandra), on cultured chicken embryonic fibroblasts. Planta Med 2006; 72(4 Suppl):289-94. 77. Mai LM, Lin CY, Chen CY, Tsai YC. Synergistic effect of bismuth subgallate and borneol, the major components of Sulbogin, on the healing of skin wound. Biomaterials 2003; 24(18 Suppl):3005-12. 78. Stevenson PC, Simmonds MS, Sampson J, Houghton PJ, Grice P. Wound healing activity of acylated iridoid glycosides from Scrophularia nodosa. Phytother Res 2002; 16(1 Suppl):33-5. 79. Riella KR, Marinho RR, Santos JS, Pereira-Filho RN, Cardoso JC, Albuquerque-Junior RL, Thomazzi SM.

Anti-inflammatory and cicatrizing activities of thymol, a

monoterpene of the essential oil from Lippia gracilis, in rodents J Ethnopharmacol 2012; 143(2 Suppl):656-63. 80. Zhang K, Qian Y, Wang H, Fan L, Huang C, Yin A, Mo X. Genipin-crosslinked silk fibroin/hydroxybutyl chitosan nanofibrous scaffolds for tissue-engineering application. J Biomed Mater Res A 2010; 95:870-81. 81. Villegas LF, Marçalo A, Martin J, Fernández ID, Maldonado H, Vaisberg AJ, Hammond GB. +)-epi-Alpha-bisabolol [correction of bisbolol] is the wound-healing principle of Peperomia galioides: investigation of the in vivo wound-healing activity of related terpenoids. J Nat Prod 2001; 64(10 Suppl):1357-9.



82. Shim KM, Choi SH, Jeong MJ, Kang SS. Effects of aucubin on the healing of oral wounds. In Vivo 2007; 21(6 Suppl.):1037-41. 83. Park TJ, Park YS, Lee TG, Ha H, Kim KT.Inhibition of acethylcholine -mediated effects by borneol. Biochem Pharmacol2003; 65(1 Suppl):83-90. 84. Annan K, Houghton,PJ . Two novel lupane triterpenoids from Paullinia pinnata L. with fibroblast stimulatory activity. J Pharm Pharmacol 2010; 62(5 Suppl):663-8. 85. Kuonen R, Weissenstein U, Urech K, Kunz M, Hostanska K, Estko M, Heusser P, Baumgartner S. Effects of Lipophilic Extract of Viscum album L. and Oleanolic Acid on Migratory Activity of NIH/3T3 Fibroblasts and on HaCat Keratinocytes. Evid Based Complement Alternat Med 2013; 718105-11. 86. Coldren CD, Hashim P, Ali JM, Oh SK, Sinskey AJ, Rha C. Gene expression changes in the human fibroblast induced by Centella asiatica triterpenoids. Planta Med 2003; 69(8 Suppl):725-32. 87. Liu M1, Dai Y, Li Y, Luo Y, Huang F, Gong Z, Meng Q. Madecassoside isolated from Centella asiatica herbs facilitates burn wound healing in mice. Planta Med 2008; 74(8 Suppl):809-15. 88. Wu F, Bian D, Xia Y, Gong Z, Tan Q, Chen J, Dai Y. Identification of Major Active Ingredients Responsible for Burn Wound Healing of Centella asiatica Herbs. Evid Based Complement Alternat Med 2012;848093-101. 89. Wright K, Rooney N, Feeney M, Tate J, Robertson D, Welham M, Ward S. Differential expression of cannabinoid receptors in the human colon: cannabinoids promote epithelial wound healing. Gastroenterology 2005; 129(2 Suppl):437-53. 90. Ben-Shabat S, Hanus LO, Katzavian G, Gallily R.

New cannabidiol derivatives:

synthesis, binding to cannabinoid receptor, and evaluation of their antiinflammatory activity. J Med Chem 2006; 49(3 Suppl):1113-7.


Page 28 of 47

Page 29 of 47


91. Drozgyik A, Hegedüs V, Lotz G, P Ónody P, A Szijártó A, A Blázovics A, K Ditrói K. Effect of different dose cannabidiol treatment on experimental induced colitis. Z Gastroenterol 2011; 49:12. 92. Dembitsky VM. A Chemistry and biodiversity of the biologically active natural glycosides. Chemistry and biodiversity2004; 1: 673-778. 93. Lindhorst, TK. Essentials of Carbohydrate Chemistry and Biochemistry. Wiley-VCH, 2007. 94. Clericuzio M, Burlando B, Gandini G, Tinello S, Ranzato E, Martinotti S, Cornara L. Keratinocyte wound healing activity of galactoglycerolipids from the fern Ophioglossum vulgatum L. J Nat Med2014;68(1 Suppl):31-7. 95. Kim YS, Cho IH, Jeong MJ, Jeong SJ, Nah SY, Cho YS, Kim SH, Go A, Kim SE, Kang SS, Moon CJ, Kim JC, Kim SH, Bae CS. Therapeutic effect of total ginseng saponin on skin wound healing. J Ginseng Res2011;35(3 Suppl):360-7. 96. Czemplik M1, Kulma A, Bazela K, Szopa J. The biomedical potential of genetically modified flax seeds overexpressing the glucosyltransferase gene. BMC Complement Altern Med 2012; 10:251-9. 97. Esposito D, Munafo JP Jr, Lucibello T, Baldeon M, Komarnytsky S, Gianfagna TJ. Steroidal glycosides from the bulbs of Easter lily (Lilium longiflorum Thunb.) promote dermal fibroblast migration in vitro. J Ethnopharmacol 2013;148(2 Suppl):433-40. 98. Munafo JP, Ramanathan A, Jimenez LS, Gianfagna TJ.

Isolation and structural

determination of steroidal glycosides from the bulbs of easter lily (Lilium longiflorum Thunb.) J Agric Food Chem2011;58(15 Suppl):8806-13. 99. Kurisu M, , Nakasone R, Miyamae Y, Matsuura D, Kanatani H, Yano S, Shigemori H. Induction of hepatocyte growth factor production in human dermal fibroblasts by caffeic acid derivatives. Biol Pharm Bull 2013; 36(12 Suppl):2018-21.



100.

Page 30 of 47

Funes L, Laporta O, Cerdán-Calero M, Micol V. Effects of verbascoside, a

phenylpropanoid glycoside from lemon verbena, on phospholipid model membranes. Chem Phys Lipids 2010; 163(2 Suppl):190-9. 101.Korkina LG, M ikhal'chik E, Suprun MV, Pastore S, Dal Toso R.

Molecular

mechanisms underlying wound healing and anti-inflammatory properties of naturally occurring biotechnologically produced phenylpropanoid glycosides. Cell Mol Biol 2007;53(5):84-91. 102. Backhouse N, Delporte C, Apablaza C, Farías M, Goïty L, Arrau S, Negrete R, Castro C, Miranda H. Antinociceptive activity of Buddleja globosa (matico) in several models of pain. J Ethnopharmacol 2008;119(1 Suppl):160-5. 103. Kuhn M, von Mering C, Campillos M, Jensen LJ, Bork P. STITCH: interaction networks of chemicals and proteins. Nucleic Acids Res 2008;36: 684-8. 104. Kuhn M, Szklarczyk D, Franceschini A, Campillos M, von Mering C, Jensen LJ, Beyer A, Bork P. STITCH 2: an interaction network database for small molecules and proteins. Nucleic Acids Res 2010; 38: 552-6. 105. Kuhn M, Szklarczyk D, Pletscher-Frankild S, Blicher TH, von Mering C, Jensen LJ, Bork P. STITCH 4: integration of protein-chemical interactions with user data. Nucleic Acids Res 2014; 42: 401-7. 106. Yanai H, Budovsky A, Tacutu R, Fraifeld VE. Is rate of skin wound healing associated with aging or longevity phenotype? Biogerontology 2011; 12(6 Suppl.):591-7. 107. Wound healing and fibrosis-related genes. http://www.resolve-whfg.appspot.com. 108. Bahramsoltani R, Farzaei MH, Rahimi R. Medicinal plants and their natural components as future drugs for the treatment of burn wounds: an integrative review. Arch Dermatol Res. 2014; 306(7 Suppl.):601-17.


Page 31 of 47


109. Molazem Z, Mohseni F, Younesi M, Keshavarzi S. Aloe vera gel and cesarean wound healing; a randomized controlled clinical trial. Glob J Health Sci. 2014; 7(1 Suppl):37955. 110.Eqhdampour F, Jahdie F, Kheyrkhah M, Taghizadeh M, Naghizadeh S, Hagani H. The Impact of Aloe vera and Calendula on Perineal Healing after Episiotomy in Primiparous Women: A Randomized Clinical Trial. J Caring Sci. 2013;2(4 Suppl.):279-86. 111. Dutta S, Bhattacharyya D. Enzymatic, antimicrobial and toxicity studies of the aqueous extract of Ananas comosus (pineapple) crown leaf. J Ethnopharmacol 2013; 150(2 Suppl.):451-7. 112. Fikru A, Makonnen E, Eguale T, Debella A, Abie Mekonnen G. Evaluation of in vivo wound healing activity of methanol extract of Achyranthes aspera L. J Ethnopharmacol.2012; 143(2 Suppl):469-74. 113. Barua CC, Talukdar A, Begum SA, Pathack DC, Sarma DK, Borah RS, Gupta A (2012) In vivo wound-healing efficacy and antioxidant activity of Achyranthes aspera in experimental burns. Pharm. Boil 2012; 50 (7 Suppl): 892-9. 114 . Jain N, Jain R, Jain A, Jain DK, Chandel HS. Evaluation of wound-healing activity of Acorus calamus Linn. Nat Prod Res 2010; 24(6 Suppl):534-41. 115. Shi GB, Wang B, Wu Q, Wang TC, Wang CL, Sun XH, Zong WT, Yan M, Zhao QC, Chen YF, Zhang W. Evaluation of the wound-healing activity and anti-inflammatory activity of aqueous extracts from Acorus calamus L. Pak J Pharm Sci 2014;27(1 Suppl):91-5. 116. Nayak S, Nalabothu P, Sandiford S, Bhogadi V, Adogwa A. Evaluation of wound healing activity of Allamanda cathartica. L. and Laurus nobilis L. extracts on rats. BMC Complement Altern Med 2006; 6:12. 117. Barua CC, Begum SA, Barua AG, Borah RS, Lankar M .Anxyolytic and anticonvulsant activity of methanol extract of leaves of Alternanthera brasiliana (L)



Kuntze (Amaranthaceae) in laboratory animals. Indian J. Pharmacol 2013; 44 (6 Suppl): 694-8. 118. Baru CC, Talukdar A, Begum SA, Buragohain B, Roy JD, Pathak DC, Sarma DK, Gupta AK, Bora RS. Effect of Alternanthera brasiliana (L) Kuntze on healing of dermal burn wound. Indian J. Exp. Biol 2012;50 (1 Suppl.): 56- 60. 119. Gal P, Vasilenko T, Kováč I, Kostelníková M, Jakubčo J, Szabo P, Dvořánková B, Sabol F, Gabius HJ, Smetana KJ Atropa belladonna L. water extract: modulator of extracellular matrix formation in vitro and in vivo. Physiol.Res 2012; 61 (3 Suppl.): 24150. 120. Parente LM,. Andrade MA, Brito LA, Moura VM, Miguel MP, Lino-Júnior Rde S, Tresvenzol LF, Paula JR, Paulo NM. Angiogenic activity of Calendula officinalis flowers L. in rats. Acta Cir Bras 2011; 26(1 Suppl):19-24. 121. Parenthe LM, Lino Júnior Rde S, Tresvenzol LM, Vinaud MC, de Paula JR, Paulo NM

Wound Healing and Anti-Inflammatory Effect in Animal Models of Calendula

officinalis L. Growing in Brazil. Evid Based Complement Alternat Med 2012; 375671:110. 122. Sanwal R, Chaudhary AK. Wound healing and antimicrobial potential of Carissa spinarum Linn. in albino mice. J Ethnopharmacol 2011 ;135(3 Suppl.):792-6. 123. Sharma Y, Jeyabalan J, Singh R.Potential wound healing agents from medicinal plants: a review. Pharmacologia 2013; 4 (5 Suppl): 1-10. 124. Harishkumar M, Masatoshi Y, Hiroshi S, Tsuyomu I, Masugi M. Revealing the mechanism of in vitro wound healing properties of Citrus tamurana extract. Biomed. Res. Int. 2013; 963457: 1-10. 125. Priva KS, Gnanamani A, Radhakrishnan N, Babu M. Healing potential of Datura alba on burn wounds in albino rats. J Ethnopharmacol 2009; 83(3 Suppl):193-9.


Page 32 of 47

Page 33 of 47


126. Bhaskar A, Nithya V. Evaluation of the wound-healing activity of Hibiscus rosa sinensis L (Malvaceae) in Wistar albino rats. Indian J. Exp. Biol 2012;51 (6 Suppl): 450-7. 127. Khalil EA, Afifi FU, Al-Hussaini M. Evaluation of the wound healing effect of some Jordanian traditional medicinal plants formulated in Pluronic F127 using mice (Mus musculus) J Ethnopharmacol 2007;109 (1 Suppl):104-12. 128. Agyare C, Bempah SB, Boakye YD, Avande PG, Adarkwa-Yiadom M, Mensah KB. Evaluation of antimicrobial and wound healing potential of Justicia flava and Lannea welwitschii. Evid. Based Complement Alternat Med 2013;63927: 1-70. 129. Schmidt C, Fronza M, Goettert M, Geller F, Luik S, Flores EM, Bittencourt CF, Zanetti GD, Heinzmann BM, Laufer S, Merfort I. Biological studies on Brazilian plants used in wound healing. J Ethnopharmacol 2009;122(3 Suppl):523-32. 130. Nayak SB, Rodrigues V, Maharaj S, Bhogadi VS. (2013) Wound healing activity of the fruit skin of Punica granatum. J Med Food.;16(9):857-861. 131. Roy P, Amdekar S, Kumar A, Singh R, Sharma P, Singh V. In vivo antioxidative property, antimicrobial and wound healing activity of flower extracts of Pyrostegia venusta (Ker Gawl) Miers. J Ethnopharmacol 2012; 140(1 Suppl):186-92. 132. Lodhi S, Pawar RS, Jain AP, Singhai AK. Wound healing potential of Tephrosia purpurea (Linn.) Pers. in rats. J Ethnopharmacol 2006;108(2 Suppl):204-10. 133. Panda V,

Thakur T. Wound Healing Activity of the Inflorescence of Typha

elephantina (Cattail). Int J Low Extrem Wounds 2013; 13(1 Suppl):50-7. 134. Sarmento Pde A, Ataíde Tda R, Pinto AP, Araújo-Júnior JX, Lúcio IM, Bastos ML. Evaluation of the extract of Zeyheria tuberculosa with a view to products for wound healing .Rev Lat Am Enfermagem 2014; 22(1 Suppl):165-72.



135. Somanath PR1, Chen J, Byzova TV. Akt1 is necessary for the vascular maturation and angiogenesis during cutaneous wound healing. Angiogenesis 2008; 11(3 Suppl):27788. 136. Low QE, Drugea IA, Duffner LA, Quinn DG, Cook DN, Rollins BJ, Kovacs EJ, DiPietro LA. Wound healing in MIP-1alpha(-/-) and MCP-1(-/-) mice. Am J Pathol 2001; 159(2 Suppl ):457-63. 137. Beare AH, O'Kane S, Krane SM, Ferguson MW. Severely impaired wound healing in the collagenase-resistant mouse. J Invest Dermatol 2003; 120(1 Suppl):153-63. 138. Hansen LA, Alexander N, Hogan ME, Sundberg JP, Dlugosz A, Threadgill DW, Magnuson T, Yuspa SH. Genetically null mice reveal a central role for epidermal growth factor receptor in the differentiation of the hair follicle and normal hair development.Am. J.Pathol 1997;150 (6 Suppl): 1959-75. 139. Kapoor M, Liu S, Shi-wen X, Huh K, McCann M, Denton CP, Woodgett JR, Abraham DJ, Leask A. GSK-3beta in mouse fibroblasts controls wound healing and fibrosis through an endothelin-1-dependent mechanism. J Clin Invest 2008; 118(10 Suppl):3279-90. 140. Owings RA, Boerma M, Wang J, Berbee M, Laderoute KR, Soderberg LS, Vural E, Jensen MH. Selective deficiency of HIF-1alpha in myeloid cells influences secondary intention wound healing in mouse skin.In Vivo 2009; 23(6 Suppl):879-84. 141. Gallucci RM, Lee EG, Tomasek JJ. IL-6 modulates alpha-smooth muscle actin expression in dermal fibroblasts from IL-6-deficient mice. J Invest Dermatol 2006; 126(3 Suppl):561-8. 142. Sano S, Itami S, Takeda K, Tarutani M, Yamaguchi Y, Miura H, Yoshikawa K, Akira S, Takeda J. Keratinocyte-specific ablation of Stat3 exhibits impaired skin


Page 34 of 47

Page 35 of 47


remodeling, but does not affect skin morphogenesis. EMBO J.1999; 18(17 Suppl):465768. 143. Shinozaki M, Okada Y, Kitano A, Ikeda K, Saika S, Shinozaki M. Impaired cutaneous wound healing with excess granulation tissue formation in TNFalpha-null mice. Arch Dermatol Res 2009; 301(7 Suppl):531-7. 144. Tyner SD, Venkatachalam S, Choi J, Jones S, Ghebranious N, Igelmann H, Lu X, Soron G, Cooper B, Brayton C, Park SH, Thompson T, Karsenty G, Bradley A, Donehower LA. p53 mutant mice that display early ageing-associated phenotypes. Nature 2002; 415:45-53.



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Table 1 WH properties of plant extracts Plant (Scientific name) Achyranthes aspera L

Plant organs

Type extract

Leaves

Methanol

Acorus calamus Leaves L

Aqueous

Allamanda cathartica L Alternanthera brasiliana L

Leaves

Aqueous

Leaves

Methanol

Atropa belladonna L

Overground parts

Aqueous

Calendula officinalis L Carissa spinarum L Cedrus deodara Roxb Citrus tamurana Hort ex Datura alba L Hibiscus rosa sinensis L

Flowers

Ethanol

Root

Ethanol

Overground parts

Hexane

Peel

Aqueous

Leaves

Ethanol

Flowers

Ethanol

Overground parts? Leaves

Aqueous

Inula viscosa L Justicia flava L

Methanol

of Mechanism action

of References

Up-regulated expression of matrix metalloproteinases (MMP-2 and 9) Inhibition of the expression of inflammatory mediators at the mRNA level Increased collagen turnover Collagen deposition, fibroblast proliferation, angiogenesis, and development of basement membrane Induction of fibronectin and galectin-1 rich ECM formation in vitro and in vivo. Angiogenesis

(112, 113)

(114, 115)

(116) (117, 118)

(119)

(120, 121)

Increasing collagen level Unknown

(122)

Transcriptional regulation of fibroblasts Expression MMP9 Increase DNA, total protein and total collagen Unknown

(124)

Improving angiogenesis,


(123)

(125) (126)

(127) (128)

Page 37 of 47


Lannea welwitschii (Hiern) Engl.

Leaves

Methanol

Parietaria diffusa Mert. & Petiveria alliacea L

Overground parts? Overground parts?.

Aqueous

Punica granatum L

Fruit skin

Pyrostegia venusta Miers

Flowers

Rubus sanctus Overground Schreber parts

Schinus L

molle Overground parts

Tephrosia purpurea L

Overground parts

Typha elephantine Roxb Watheria douradinha Saint-Hilaire

Flowers

Zeyheria tuberculosa Buman

Overground parts.

Stems

collagenation and reepithelialization Improving (128) angiogenesis, collagenation and reepithelialization Unknown (127)

Ethanol

Decrease NF-cB activation, influence on elastase Methanol During early WH phase increasing TNF-α and IL-6 level Methanol During early WH phase increasing TNF-α and IL-6 level n-hexane, Speeding up the chloroform, proliferation ethyl acetate phase and methanol Schmidt et al, Decrease NF-cB 2009 activation, influence on elastase Ethanol Increase in fibroblast cells, collagen fibres and blood vessels formation Methanol Increasing collagen formation Ethanol, Decrease NF-kB hexane activation, influence on elastase Ethanol Unknown


(129)

(130)

(131)

(11)

(129)

(132)

(133)

(129)

(134)


Page 38 of 47

Table 2 Selected targets of pro-WH herbal compounds

Herbal compounds

Target

Relevance of the target to WH

Reference

Curcumin,

AKT1

Akt1 is associated with assembly of (135)

Epicathechin gallate

collagen in skin wounds and around the

Gallic

blood vessels. Wounds in Akt1 (-/-) mice,

acid,

Isorhamnetin,

but not in

Akt2

(-/-) mice,

were

Kaempferol, Nicotine

characterized by reduced vascular area as

Quercetin,

well as impaired vascular maturation as

Resveratrol

evidenced by reduced smooth muscle cell recruitment.

Curcumin,

CCL2

Associated

with

wound

re- (136)


epithelialization as the KO mice had

Kaemferol

impaired wound angiogenesis.

Quercetin Asiaticoside,

COL1A1

WH was severely delayed in the KO mice, (137)


Page 39 of 47


Curcumin

with

wounds

remaining

significantly

Epicathechin gallate,

wider than wild-type for the first 2 wk

Nicotine,

after injury. Re-epithelialization of the

Quercetin

Col1a1(r/r) wounds took 7 d longer than in the wild-type. The Col1a1(r/r) wounds had a prolonged early inflammatory response.

Curcumin,

EGFR


Modifies both normal and wound-induced (138) epidermal proliferation

Quercetin, Resveratrol, Silibinin Curcumin,

GSK3B

Gsk3b-conditional-KO mice (Gsk3b-CKO (139)


mice)

exhibited

accelerated

Quercetin,

closure,

Resveratrol

excessive scarring compared with control

increased

wound

fibrogenesis,

and

mice. In addition, Gsk3b-CKO mice showed elevated collagen production, decreased cell apoptosis, elevated levels of profibrotic alpha-SMA, and increased myofibroblast formation during wound healing. Curcumin,

HIF1A

Early

wound

closure

occurred (140)


significantly faster in HIF-1alpha KO

Quercetin,

mice than in WT mice. Wounds of KO

Resveratrol, Silibinin

mice

contained

similar

numbers

of

neutrophils and macrophages, but more activated keratinocytes, consistent with accelerated re-epithelialization. Curcumin,

IL6

IL-6-deficient transgenic mice (IL-6 KO) (141)

Epicatechin,

displayed significantly delayed cutaneous


wound healing compared with wild-type

Kaempferol

control animals, requiring up to threefold

Nicotine,

longer to heal. This was characterized by

Quercetin,



Resveratrol

minimal

epithelial

bridge

Page 40 of 47

formation,

decreased inflammation, and granulation tissue formation. Curcumin,

STAT3

KO mice, whose epidermal and follicular (142)


keratinocytes lack functional Stat3, were

Nicotine, Resveratrol,

viable and the development of epidermis

Silibinin

and

hair

follicles

appeared

normal.

However, hair cycle and wound healing processes were severely compromised. Curcumin,

TNF

Knockout promotes granulation tissue (143)


formation and retards re-epithelialization

Gallic acid, Nicotine, Quercetin Resveratrol, Silibinin Asiatic

acid, TRP53

The wound-healing assay showed a (144)

Curcumin,

significant reduction in wound closure for


the pL53 transgenic mice 4 days after

Nicotine,

induction of a 3-mm punch biopsy in the

Quercetin,

Resveratrol, Silibinin

dorsal skin

Table 3 Clinical trials dealing with the effect of medicinal plant products on WH. Herbal product tested

Outcome of the study

Aloe vera gel

Double-blind clinical trial 109 demonstrated

References

significant

WH effect of the gel at 24 hours post cesarean


Page 41 of 47


Aloe

vera,

ointment

Calendula One

control

and

two 110

experimental groups were investigated. of

each

Application product

significantly increased the speed of episiotomy WH.



Page 42 of 47

Figure legends Figure 1: Chemical structures of alkaloids isolated from Aconitum baikalense: (1-mesaconitine, 2-

hypaconitine, 3-songorine and 4-napelline) Figure 2: Chemical structure of Taspine - an alkaloid isolated from various plants of the Magnoliaceae family. Figure 3: Chemical structure of 4’,6,7-trimethoxyisoflavone. Figure 4: Chemical structures of Gentiopicroside, sweroside and swertiamarine isolated from Gentiana lutea Figure 5: Chemical structures of (22R,25R)-spirosol-5-en-3β-yl - O-α-L-rhamnopyranosyl-(1-2)-β-Dglucopyranosyl-(1-4)-β-D-glucopyranosides isolated from Lilium longiflorum


Page 43 of 47


Figure 1:

OCH3

HO

OH

O O

OCH3 O H3C

N

O O HO O

O

N

OAc OH O H3CO

H3CO

OH

O O

Mesaconitine

Hypaconitine

OH

O

CH2

OH

CH2

OH H3C

N

N

H3C

OH CH3

Napelline

OH CH3

Songorine



Figure 2: OCH3 O

O

CH3 N CH3 O

O

OCH3


Page 44 of 47

Page 45 of 47


Figure 3: OCH3 O H3CO

H3CO

O



Page 46 of 47

Figure 4:

CH2 O

HO

OH O

O

HO

O

O O

O HO O

O

HO

OH

OH

OH

H2C

Sweroside

Gentiopicroside

CH2

OH

HO

OH

O O

O

O OH

O

OH

Swertiamarine


O

Page 47 of 47


Figure 5: NH O

Glu

O Rham Glu


Determination of the Wound Healing Potentials of Medicinal Plants Historically Used in Ghana.

Effect of taurine on wound healing.

Effect of negative pressure wound therapy on wound healing.

Wound Healing Properties of Selected Plants Used in Ethnoveterinary Medicine.

Periodontal repair in dogs: effect of wound stabilization on healing.

Effect of Ampelopsis Radix on wound healing in scalded rats.

The effect of selective desalivation on wound healing in mice.

Effect of low power laser on incisional wound healing.

Effect of low power laser on incisional wound healing.

Effect of ultraviolet therapy on rat skin wound healing.

Effect of Propolis on Experimental Cutaneous Wound Healing in Dogs.

Effect of haematocrit on wound healing in children.

Lateral patellar release in knee arthroplasty. Effect on wound healing.

Wound healing: a review. I. The biology of wound healing.

Influence of oxygen on wound healing.

Influence of the pineal on wound healing.

Wound healing.

Wound healing.

Wound healing: a review. II. Environmental factors affecting wound healing.

Indian Medicinal Plants.

Wound healing.

Indian Medicinal Plants.

Lanna Medicinal Plant Recipe Database "MANOSROI III".

The effect of uraemia upon wound healing: an experimental study.