Linear Incision Wound Models Research Articles

Preclinical evaluation of the diabetic wound healing activity of phytoconstituents extracted from Ficus racemosa Linn. leaves

Human body has several multi-layered organs, but skin is one of biggest and easiest to access. It serves as body's primary line of defense alongside various skin diseased conditions. Despite receiving sufficient and appropriate care, diabetes wounds heal slowly and may take a week to complete. A progression of connective tissue patch up is the body's natural defense against tissue damage. Fresh leaves of Ficus racemosa were utilized for the study. In this study, two distinct models were employed to compare how well different Ficus racemosa leaf extracts healed wounds. Excision wounds healed more quickly and to a greater extent after being treated with a flavonoid and tannin fraction of Ficus racemosa leaf extract, suggesting improved epithelization. The extract-treated groups also experienced an increase in breaking potency of incision wounds made; higher breaking strength indicates better wound healing. Complete closure of wound of flavonoid fraction and in fraction of Ficus racemosa extract occurred in 16 and 17 days respectively. Standard treatment increased tensile strength in the diabetic linear incision wound model, followed by treatment with the flavonoid fraction and tannin fraction of Ficus racemosa leaves extract. Ultimate finding and outcome of the present study experimentally demonstrates that extracts of the flavonoid and tannin fractions of Ficus racemosa have wound-healing properties and are effective in treating diabetic wounds. From this study, we state that Ficus racemosa flavonoid fraction and tannin fraction extract has a beneficial effect on blood glucose levels, which shows hypoglycaemic activity.

Phytochemical Profiling and Biological Activity of Achillea sintenisii Hub.-Mor.

Achillea (Asteraceae) species have been traditionally used for their different therapeutical properties. In this study, phytochemical composition of aerial parts of A. sintenisii which is endemic in Turkey was determined with Liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS). To evaluate the wound healing potential, the cream formulation prepared from A. sintenisii was tested on the linear incision wound model in mice. In vitro enzyme inhibitory activity tests were performed on elastase, hyaluronidase, and collagenase. In the histopathological examination, angiogenesis and granulation tissue formation were significantly increased in A. sintenisii treatment groups compared to the negative control group. As a result of this study, it is thought that the enzyme inhibition and antioxidant activity of the plant may contribute to the wound healing process. According to LC/MS/MS analysis result, quinic acid (24.261 μg/mg extract) and chlorogenic acid (14.97 μg/mg extract) were identified as main constituents of the extract.

In vivo wound-healing activity of Euphorbia characias subsp. wulfenii: Isolation and quantification of quercetin glycosides as bioactive compounds

Ethnopharmacological relevanceThe latex and the aerial parts of Euphorbia characias L. (Euphorbiaceae) have been used as medicinal plant to treat wounds and warts in traditional medicine. Aim of the studyThe effect of the plant extract was tested in vivo and in vitro with experimental models to find scientific evidence for traditional use in wound healing. Potentially active wound-healer compounds were isolated from the active fraction using fractionation procedures under the guidance of biological assay and the possible role of the compounds in the wound healing process was also determined. Material and methodsN-hexane, ethyl acetate, and methanol extracts were successively prepared from the aerial parts of E. characias subsp. wulfenii. The extracts were tested with linear incision, circular excision wound models and the hydroxyproline assay method to assess the wound-healing activity. The inhibition of the increase in capillary permeability induced by acetic acid, an acute inflammation model, was used to assay the anti-inflammatory activity. Different chromatographic separation techniques on sephadex and silica gel columns, and bioassay guided assay techniques have been used to isolate the active compounds of the plant. Moreover, hyaluronidase, collagenase and elastase enzymes inhibitory effect of active principle were investigated in vitro to find out the mechanism of action. ResultsThe methanol (MeOH-ex) extract of the aerial parts of E. characias subsp. wulfenii showed significant wound healing activity (linear incision wound model: 43.04%; circular excision wound model 65.24%) and anti-inflammatory activity (34.74%). The methanol extract was separated into its fractions by column chromatography for isolation of efficient compounds. Biological activity of the fractions were assessed and further isolation and purification processes have been carried out in the active fraction. Isolation studies were carried out from the MeOH-ex fraction to obtain active constituents and their structures were elucidated to be quercetin-3-O-rhamnoside (quercitrin), quercetin-3-O-galactoside (hyperoside), and quercetin-3-O-arabinoside (guaijaverin). Further in vitro and in vivo assays showed that quercetin derivatives were responsible for the wound-healing activity of the plant, and also found to be significant anti-elastase and anti-collagenase activities. The amounts of three compounds, isolated from active fraction, were determined by using high performance liquid chromatography. Calibration equation was calculated with dilutions, prepared from pure substances, and assay was performed in total extract, prepared from E. characias subsp. wulfenii. It was detected that the plant had 1.22% quercitrin, 0.35% hyperoside, and 0.11% guaijaverin. The validation of the analytical method was performed by linearity, precision, limit of detection, and limit of quantification parameters. ConclusionPresent study supported the traditional use of the aerial parts E. characias subsp. wulfenii as wound healer and quercetin derivatives were isolated as active components from the active fraction by using bioassay-guided fractionation technique.

Gas chromatography coupled to mass spectrometry characterization, anti-inflammatory effect, wound-healing potential, and hair growth-promoting activity of Algerian Carthamus caeruleus L (Asteraceae).

OBJECTIVES:The roots of Carthamus caeruleus have been used by the population of Northern Algeria to treat several pathological conditions, including wound healing and hair growth. The present study was conducted to evaluate the anti-inflammatory activity, wound-healing potential, and hair growth-promoting activity attributed to C. caeruleus root.MATERIALS AND METHODS:In this study, we have investigated the anti-inflammatory effect using carrageenan-induced paw edema test, evaluated the wound-healing potential by linear incision wound model, and evaluated hair growth activity using in vivo hair growth-promoting test attributed to C. caeruleus root. Preliminary phytochemical screening and gas chromatography coupled to mass spectrometry (GC/MS) characterization were also performed.RESULTS:It was found that the methanolic extract of C. caeruleus was characterized by the presence of tannins, flavonoids, anthocyanins, leucoanthocyanins, sennosides, free quinones, saponins, glycosides, mucilage, and coumarins. The GC/MS analysis could identify 22 compounds and showed that the major chemical constituents were palmitic acid (12.88%), mono(2-ethylhexyl) phthalate (12.75%), and 5-(hydroxymethyl)-2-furancarboxaldehyde, (9.19%). The phytoextract strongly inhibited (P < 0.001) paw edema formation in mice. The roots of C. caeruleus also showed a significant (P < 0.05) wound-healing and hair growth-promoting effects.CONCLUSION:The results indicate the richness of the roots of the Algerian C. caeruleus in biomolecules. These molecules exhibit an excellent reducing inflammation activity, a wound-healing property, and an interesting hair-promoting growth activity. All in all, the findings promote the usage of the Algerian C. caeruleus as an effective and a safe potential skincare alternative remedy.

Isoflavonoids as wound healing agents from Ononidis Radix

Ethnopharmacological relevanceDried roots of Ononis spinosa L. are traditionally used for their diuretic, anti-inflammatory and wound healing effects. Aim of the studyIsolation of the bioactive compounds of Ononis spinosa L. subsp. leiosperma (Boiss.) Sirj. Materials and methodsEthyl acetate extract prepared from the roots of Ononis spinosa L. subsp. leiosperma (Boiss.) Sirj. was subjected to silica gel column. The fractions were tested for their wound healing and anti-inflammatory activities. Linear incision and circular excision wound models and hydroxypyroline estimation assay were used for the wound healing activity. Carrageenan-induced hind paw edema, TPA-induced ear edema and acetic acid-induced increase in capillary permeability tests as acute inflammation; FCA-induced arthritis as chronic inflammation models were used for the assessment of anti-inflammatory activity. Antioxidant capacities of the fractions were tested using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid (ABTS) scavenging activity assay, reducing power assay and hydroxyl radical (OH-) scavenging assay. The isolation procedure was continued with the active fraction (Fr-E5). ResultsFr-E5 exhibited remarkable wound healing activity with the 33.4% tensile strength value on the linear incision wound model and 51.4% reduction of the wound area at the day 12 on the circular excision wound model. Hydroxyproline content of the tissue treated by Fr-E5 was found to be 30.9 ± 0.72μg/mg. Acetic acid induced increase in capillary permeability test results revealed that Fr-E5 inhibited inflammation by the value of 40.3%. Fr-E5 showed 28.1–32.2% inhibition in carrageenan-induced hind paw edema test while did not possess activity on TPA-induced ear edema and FCA-induced arthritis models. Trifolirhizin, ononin, medicarpin-3-O-glucoside, onogenin-7-O-glucoside and sativanone-7-O-glucoside were isolated from Fr-E5 and tested for their wound healing activities using by measuring their inhibition of hyaluronidase, collagenase and elastase enzymes. Ononin and sativanone-7-O-glucoside inhibited hyaluronidase and elastase enzymes by 31.66% and 41.75%; 45.58% and 46.88% values respectively at the dose of 100μg/mL. ConclusionAmong five isolated compounds, ononin and sativanone-7-O-glucoside were found to inhibit hyaluronidase and elastase enzymes. According to the results, these compounds may majorly be responsible for the wound healing activity of the extract.

Wound healing and anti-inflammatory activity of some Ononis taxons

Ononis species are used for their laxative, diuretic, analgesic, anti-inflammatory, antiviral, cytotoxic and antifungal effects as well as against skin diseases for wound healing activity. In the light of this information n-hexane, ethylacetate and methanol extracts prepared from Ononis spinosa L. subsp. leiosperma (Boiss.) Sirj., Ononis variegata L., Ononis viscosa L. subsp. brevifolia (DC) Nym. and Ononis natrix L. subsp. natrix L. were tested for their wound healing, anti-inflammatory and antioxidant activities. Linear incision and circular excision wound models and hydroxypyroline estimation assay were used for the wound healing activity. For the assessment of chronic inflammation FCA-induced arthritis and for acute inflammation carrageenan-induced hind paw edema, TPA-induced ear edema and acetic acid-induced increase in capillary permeability tests were conducted. 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) scavenging activity assay, reducing power assay and hydroxyl radical (OH−) scavenging assay were used for determining antioxidant activities of the extracts. Results showed that O. spinosa subsp. leiosperma roots ethyl acetate extract exhibited remarkable wound healing activity with the 42.6% tensile strength value on the linear incision wound model and 60.1% reduction of the wound area at the day 12 on the circular excision wound model. Hydroxyproline content of the tissue treated by O. spinosa subsp. leiosperma roots ethyl acetate extract was found to be 41.3μg/mg. Acetic acid induced increase in capillary permeability test results revealed that O. spinosa subsp. leiosperma roots ethyl acetate extract and O. spinosa subsp. leiosperma roots methanol extract inhibited inflammation by 40.4% and 35.4% values respectively. O. spinosa subsp. leiosperma roots ethyl acetate extract showed 21.2-27.2% inhibition in carrageenan-induced hind paw edema test while did not posses activity on TPA-induced ear edema and FCA-induced arthritis models.

Anti-Inflammatory and Wound-Healing Potential of Golden Chanterelle Mushroom, Cantharellus cibarius (Agaricomycetes).

The golden chanterelle mushroom, Cantharellus cibarius, is an edible mushroom with medicinal value. Given that this species has good radical scavenging activity and strong antioxidant potential and bactericidal effects, this study was designed to investigate the anti-inflammatory and wound-healing activity of C. cibarius extract in rats. For this experimental study, circular excision and linear incision wound models were used in 4 groups of male Wistar rats: nontreated, vehicle-treated, treated with C. cibarius extract ointment (2% w/w), and treated with the reference drug (Madecassol). All the animals were treated topically once a day. The circular and linear wounds were treated for 9 and 17 days, respectively. At the end of the study, samples from healing wounds were taken for histopathological assessment to determine the in vivo anti-inflammatory activity and investigate immunohistochemistry by cyclooxygenase-2 (COX-2). Significant wound-healing activity in each wound model was observed in the C. cibarius extract-treated and Madecassol-treated groups compared with the nontreated and vehicle-treated groups (P < 0.05). Histological assessment showed complete repair of the epidermal layer, increased collagen production, and a remarkable degree of neovascularization and epithelization in the extract group, which were significantly different from those in the other groups (P < 0.05). In addition, a significantly lower rate of COX-2 expression was detected in the extract group than in the nontreated and vehicle-treated groups (P < 0.0001). Therefore, the experimental data reveal that C. cibarius extract showed significant wound-healing and anti-inflammatory effects, which could be the scientific rationale for the medicinal use of the golden chanterelle mushroom in treating wounds.

Evaluation of wound healing property of Caesalpinia mimosoides Lam.

Ethnopharmacological relevanceCaesalpinia mimosoides Lam. is one of the important traditional folk medicinal plants in the treatment of skin diseases and wounds used by healers of Uttara Kannada district of Karnataka state (India). However scientific validation of documented traditional knowledge related to medicinal plants is an important path in current scenario to fulfill the increasing demand of herbal medicine. Aim of the studyThe study was carried out to evaluate the claimed uses of Caesalpinia mimosoides using antimicrobial, wound healing and antioxidant activities followed by detection of possible active bio-constituents. Materials and methodsExtracts prepared by hot percolation method were subjected to preliminary phytochemical analysis followed by antimicrobial activity using MIC assay. In vivo wound healing activity was evaluated by circular excision and linear incision wound models. The extract with significant antimicrobial and wound healing activity was investigated for antioxidant capacity using DPPH, nitric oxide, antilipid peroxidation and total antioxidant activity methods. Total phenolic and flavonoid contents were also determined by Folin–Ciocalteu, Swain and Hillis methods. Possible bio-active constituents were identified by GC-MS technique. RP-UFLC-DAD analysis was carried out to quantify ethyl gallate and gallic acid in the plant extract. ResultsPreliminary phytochemical analysis showed positive results for ethanol and aqueous extracts for all the chemical constituents. The ethanol extract proved potent antimicrobial activity against both bacterial and fungal skin pathogens compared to other extracts. The efficacy of topical application of potent ethanol extract and traditionally used aqueous extracts was evidenced by the complete re-epithelization of the epidermal layer with increased percentage of wound contraction in a shorter period. However, aqueous extract failed to perform a consistent effect in the histopathological assessment. Ethanol extract showed effective scavenging activity against DPPH and nitric oxide free radicals with an expressive amount of phenolic and moderate concentration of flavonoid contents. Ethyl gallate and gallic acid were found to be the probable bio-active compounds evidenced by GCMS and RP-UFLC-DAD analysis. ConclusionThe study revealed the significant antimicrobial, wound healing and antioxidant activities of tender parts of C. mimosoides and proved the traditional folklore knowledge.

Bioassay-guided isolation of wound healing active compounds from Echium species growing in Turkey

Ethnopharmacological relevanceThe roots and root barks of Echium sp. have been used to treat ulcers, burns and wounds in traditional Turkish medicine. Aim of the studyOn the basis of them traditional use and literature references, four Echium species were selected for evaluation of them wound healing potential. Isolation of active component(s) from the active extracts through the bioassay guided fractionation procedures. Material and methodsIn vivo the wound healing activity of the plants was evaluated by linear incision experimental models. The chloroform extract of Echium italicum L. was fractionated by successive chromatographic techniques. Wound healing activity of each fraction was investigated following the bioassay-guided fractionation procedures. Moreover, the tissue samples of isolated compounds were examined histopathologically. The healing potential was comparatively assessed with a reference ointment Madecassol®, which contains 1% extract of Centella asiatica. ResultsSignificant wound healing activity was observed from the ointment prepared with ethanol extract at 1% concentration. The ethanol root extract treated in groups of animals showed a significant increase (37.38%, 40.97% and 35.29% separately for E. italicum L, Echium vulgare L. and Echium angustifolium Miller) wound tensile strength in the incision wound model. Subfractions showed significant but reduced wound healing activity on in vivo wound models. Shikonin derivatives “Acetylshikonin”, “Deoxyshikonin” and “2-methyl-n-butyrylshikonin+Isovalerylshikonin”, were isolated and determined as active components of active final subfraction from E. italicum L. roots. The results of histopathological examination supported the outcome of linear incision wound models. ConclusionThe experimental study revealed that Echium species display remarkable wound healing activity.

Favorable effects of superficially deacetylated chitin nanofibrils on the wound healing process

Previous reports indicate that the beneficial effect of chitin nanofibrils (CNFs), and chitosan nanofibrils (CSNFs) for wound healing. In this study, the wound healing effects of superficially deacetylated chitin nanofibrils (SDACNFs) were evaluated using an experimental model. In the experiments using circular excision wound model, SDACNFs induced re-epithelium and proliferation of the fibroblasts and collagen tissue. In the chitin, CNFs, and CSNFs, on the other hand, the e-epithelium and proliferation of the fibroblasts and collagen tissue were not induced perfectly compared with the SDACNFs group. In particular, re-epithelization was observed on day 4 in the only SDACNF group. Moreover, SDACNFs did not induce severe systemic inflammation in the linear incision wound model. The data indicated that SDACNFs effectively induced the proliferation and re-modeling phases compared with chitin, CNFs, and CSNFs in the wound. These data indicate that SDACNFs can be beneficial as a new biomaterial for wound healing.

Evaluation of the wound healing activity of Buxus sempervirens L. and Centaurium erythraea (L.) Rafn. by using in vivo and in vitro Methods

Buxus sempervirens L. (Buxaceae) and Centaurium erythraea (L.) Rafn. (Gentianaceae) have been used for wound healing in Turkish traditional medicine. In order to evaluate the wound healing potential of these plants three different extracts were successively prepared from 500 gr of plant materials by extracting with 4000 ml of n-hexane, ethyl acetate and methanol at room temparature for 24 hours. The extracts were evaporated to dryness in vacuo. The healing process was investigated with a circular excision and a linear incision wound model to analyse the wound contraction and tensile strength in vivo. The anti-inflammatory potential, which is related to the wound healing activity, was evaluated by an in vivo experimental model based on the inhibition of acetic acid-induced increase in capillary permeability assessment. Biochemical and histopathological evaluations were done too. For the antioxidant activity assessment DPPH (2,2-diphenyl-1-picrylhydrazyl) assay was applied. According to the results of the present study, methanol extracts of both B. sempervirens and C. erythraea displayed remarkable wound healing activity with the significant tensile strength values of 30.4 and 34.7%, respectively on the linear incision wound model. Similarly, methanol extracts demonstrated notable wound healing effect with the 59.5 and 71.5% contraction values, respectively on the circular excision wound model.

Investigating wound healing, tyrosinase inhibitory and antioxidant activities of the ethanol extracts of Salvia cryptantha and Salvia cyanescens using in vivo and in vitro experimental models

Salvia L. species are widely used against wounds and skin infections in Turkish folk medicine. The aim of the present study is to evaluate wound healing activity of the ethanol (EtOH) extracts of Salvia cryptantha and Salvia cyanescens. For the assessment of wound healing activity linear incision and circular excision wound models were employed on rats and mice. The wound healing effect was comparatively evaluated with the standard skin ointment Madecassol(®). Inhibition of tyrosinase, a key enzyme in skin aging, was achieved using ELISA microplate reader. Antioxidant activity was evaluated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and superoxide radical scavenger effect, ferrous ion-chelating ability, and ferric-reducing antioxidant power (FRAP) tests. The EtOH extract of Salvia cryptantha treated groups of animals showed 56.5% contraction, whereas the reference drug Madecassol(®) showed 100% contraction. On the other hand, the same extract on linear incision wound model demonstrated a significant increase (33.2%) in wound tensile strength as compared to other groups. The results of histopathological examination maintained the upshot of linear incision and circular excision wound models as well. These findings specify that Salvia cryptantha for wound healing activity can be appealed further phytochemical estimation for spotting its active components.