Anti-inflammatory,wound healing and in-vivo antioxidant properties of the leaves of Ficus amplissima Smith

Ethno pharmacological relevance

Ficus amplissima, commonly known as kal-itchchi, being used in folklore medicine, leaves juice is applied externally on old chronic wounds and the latex is applied on the wound.

Aim

There was no scientific evidence justifying the medicinal use of Ficus amplissima, therefore the present study was aimed at evaluation of anti-inflammatory, wound healing and in-vivo antioxidant activity of the plant.

Materials and methods

In the present study the acetone extract of Ficus amplissima leaves were studied for enzymatic antioxidant activity through different assays, anti-inflammatory by using carrageenan and histamine induced rat paw edema models (50 mg, 100 mg acetone extract) and wound healing activity by incorporating the two doses (1% (w/w) and 2% (w/w)) of acetone extract and simple ointment base B.P. in concentration of 0.5% (w/w) using excision and incision wound models in rats. In case of the excision wound model wound contraction and period of epithelization was studied while incision wound model was evaluated by determining tensile strength.

Results

Ficus amplissima leaves expressed the potent anti-inflammatory and in-vivo antioxidant activity where 100 mg acetone extract showed high activity. Treatment of wound with ointment containing 2% (w/w) acetone extract exhibited significant (P<0.001) wound healing activity.

Conclusion

The acetone extract of Ficus amplissima leaves exhibited better anti-inflammatory, wound healing and in-vivo antioxidant activity probably due to phenols constituents.     

In vivo anti-inflammatory and wound healing activities of Centaurea iberica Trev. ex Spreng

Aim of the study

Several Centaurea species (Asteraceae) are used in Turkish folk medicine to alleviate pain and inflammatory symptoms in rheumatoid arthritis, high fever, head ache and for wound healing. Particularly, the aerial part of Centaurea iberica Trev. ex Spreng. has been practiced on wounds for healing. In order to evaluate the anti-inflammatory and wound healing activities of the plant, extracts were prepared with variety of solvents: hexane, chloroform, ethyl acetate and aqueous methanol (85%) from the aerial parts of Centaurea iberica.

Materials and methods

The incision by using tensiometer and excision models were used in order to assess the effect of the plant extracts on wound healing in mice and rats. Results were also evaluated histopathologically. In vivo inhibitory effect of the extracts on acetic acid-induced increase in capillary permeability was studied for the assessment of anti-inflammatory activity.

Results

The wound healing effect was comparatively evaluated with a reference ointment Madecassol^®. Noteworthy wound healing activity was observed for the ointment formulation prepared with 1% methanol extract. The results of histopathological evaluation supported the outcome of both incision and excision wound models. Moreover, the methanol extract exerted remarkable wound healing activity and also demonstrated a significant and dose-dependent anti-inflammatory activity.

Conclusion

The experimental study revealed that Centaurea iberica displays remarkable wound healing and anti-inflammatory activity.     

Wound healing activity of Calotropis gigantea root bark in rats

Ethnopharmacological relevance

Calotropis gigantea R.Br. (Asclepiadaceae) is a perennial undershrub found chiefly in wastelands throughout India. It has been reported as a traditional folkloric medicine for a variety of alignments. The plant Calotropis gigantea is also used in some parts of India for wound healing in combination with other plants. However there are no scientific reports on wound healing activity of the plant Calotropis gigantea R.Br.

Aim of the study

To investigate the effects of Calotropis gigantea root bark on wound healing activity in rats by excision, incision and dead space wound healing models in rats.

Methodology

Wistar albino rats of either sex weighing between 180 and 200 g were topically treated with extract formulated in ointment by using simple ointment BP as base. 5% (w/w) ointment was applied once daily in excision wound model. Calotropis gigantea ethanolic extract was given orally at a dose of 100, 200, and 400 mg/kg in incision and dead space wound healing models. Rats of standard groups were treated with 5% Povidone iodine ointment topically. The percentage wound closure, epithelization time, hydroxyproline content and scar area on complete epithelization were measured.

Results

Topical application of Calotropis gigantea in excision wound model increased the percentage of wound contraction. Scar area and epithelization time were decreased. In incision wound and dead space wound breaking strength of wounds and hydroxyproline was increased.

Conclusion

Calotropis gigantea accelerated wound healing in rats and thus supports its traditional use.     

In vivo wound healing activity and phytochemical screening of the crude extract and various fractions of Kalanchoe petitiana A. Rich (Crassulaceae) leaves in mice

Ethnopharmacological relevance

The leaves of Kalanchoe petitiana A. Rich (Crassulaceae) are used in Ethiopian folk medicine for treatment of evil eye, fractured surface for bone setting and several skin disorders including for the treatment of sores, boils, and malignant wounds.

Aim of the study

In order to scientifically prove the claimed utilization of the plant, the effects of the extracts and the fractions were investigated using in vivo excision, incision and dead space wound models.

Materials and method

Mice were used for wound healing study, while rats and rabbit were used for skin irritation test. For studying healing activity 80% methanolic extract and the fractions were formulated in strength of 5% and 10%, either as ointment (hydroalcoholic extract, aqueous and methanol fractions) or gel (chloroform fraction). Oral administration of the crude extract was used for dead space model. Negative controls were treated either with simple ointment or sodium carboxyl methyl cellulose xerogel, while positive controls with nitrofurazone (0.2 w/v) skin ointment. Negative controls for dead space model were treated with 1% carboxy methyl cellulose. Parameters, including rate of wound contraction, period of complete epithelializtion, hydroxyproline contents and skin breaking strength were evaluated.

Results

Significant wound healing activity was observed with ointment formulated from the crude extract at both 5% and 10% concentration (p<0.01) compared to controls in both excision and incision models. In dead space model, 600 mg/kg (p<0.01) but not 300 mg/kg significantly increased hydroxyproline content. Fractions showed variable effect, with the chloroform fraction lacking any significant effect. Both 5% and 10% formulations of the aqueous and methanolic fractions significantly increased wound contraction, decreased epithelializtion time and increased hydroxyproline content in excision wound model (p<0.05) as compared to controls. These fractions were also endowed with higher skin breaking strength in incision wound model (p<0.01).

Conclusions

The present study provided evidence that the leaves of Kalanchoe petitiana A. Rich possess remarkable wound healing activities supporting the folkloric assertion of the plant. Fractionation revealed that polar or semi polar compound may play vital role, as both aqueous and methanolic fractions were endowed with wound healing activity.     

Evaluation of wound healing activity of root of Mimosa pudica

Ethno pharmacological relevance

Mimosa pudica, commonly known as touch-me-not, is used in folklore medicine in arresting bleeding and in skin diseases.

Aim

There was no scientific evidence justifying the use of Mimosa pudica, therefore the present study was aimed at evaluation of wound healing activity of the plant.

Materials and methods

In the present study the roots of Mimosa pudica were studied for wound healing activity by incorporating the methanolic and the total aqueous extract in simple ointment base B.P. in concentration of 0.5% (w/w), 1% (w/w) and 2% (w/w). Wound healing activity was studied in three types of model in rats viz. excision, incision and estimation of biochemical parameter. In case of the excision wound model wound contraction and period of epithelization was studied while in incision wound model was evaluated by determining tensile strength and hydroxyproline content in the scab.

Results

Treatment of wound with ointment containing 2% (w/w) the methanolic and 2% (w/w) the total aqueous extract exhibited significant (P < 0.001) wound healing activity. The methanolic and total aqueous extracts were analyzed for total phenols content equivalent to Gallic acid. The content of total phenols was 11% (w/w) and 17% (w/w) in methanolic and total aqueous extract respectively.

Conclusion

The methanolic extract exhibited good wound healing activity probably due to phenols constituents.     

Exploring the wound healing activity of Arnebia densiflora (Nordm.) Ledeb. by in vivo models

Ethnopharmacological relevance

The roots of Arnebia densiflora (Nordm.) Ledeb. (Boraginaceae), which is an endemic plant for Turkey, have been used for healing of wounds in folk medicine. The extract of the roots in olive oil and mostly the root barks was applied onto open wounds for rapid healing.

Aim of the study

To evaluate the wound healing activity of the roots, extracts were prepared with different solvents; hexane, chloroform, ethyl acetate and methanol, respectively from the roots of Arnebia densiflora.

Materials and methods

Incision by using tensiometer and excision models were employed on mice and rats.

Results

Significant wound healing activity was observed with the ointment formulation prepared by using hexane extract at 1% concentration on the mentioned models. The results of histopathological examination supported the outcome of both incision and excision wound models. The wound healing effect was comparatively evaluated with a reference ointment Madecassol^®.

Conclusion

The experimental data demonstrated that Arnebia densiflora displayed remarkable wound healing activity.     

Bioactivity guided evaluation of anti-inflammatory and antinociceptive activities of Arceuthobium oxycedri (D.C.) M. Bieb.

Ethnopharmacological relevance

Arceuthobium oxycedri (D.C.) M. Bieb. (Loranthaceae) or dwarf mistletoe is a rare semi-parasitic plant that lives on the branches of Juniperus oxycedrus as the host plant. In Turkish folk medicine, the plant is prescribed as a panacea for every kind of diseases, including, against infectious and inflammatory disorders of upper respiratory system and gastro-intestinal complaints or as a hypotensive remedy.

Aim of the study

In the current study, in vivo anti-inflammatory and antinociceptive effects of Arceuthobium oxycedri have been investigated.

Material and methods

The crude ethanolic extract of the whole plant was sequentially fractionated into five subextracts; explicitly, n-hexane, chloroform, ethyl acetate (EtOAc), n-butanol, and remaining water extracts. Further studies were carried out on the most active subextract, i.e. the EtOAc subextract, was further subjected to fractionation through successive column chromatographic applications on Silica gel 60, Sephadex LH-20 and LiChropep RP-18. For the activity assessment, each extract or fraction was submitted to bioassay systems; carrageenan-induced hind paw edema model for anti-inflammatory activity and p-benzoquinone induced abdominal contraction test for antinociceptive activity assessment.

Results

Among the extracts obtained, the ethanolic extract, EtOAc and n-butanol subextracts showed significant inhibitory activity in the bioassay systems. From the EtOAc subextract, a major component was isolated and its structure was determined as (+)-catechin by means of spectral techniques.

Conclusion

Present study confirms the claimed utilization of the plant against inflammatory complaints in Turkish folk medicine.     

Biological activities of Pseudevernia furfuracea (L.) Zopf extracts and isolation of the active compounds

Ethnopharmacological relevance

Pseudevernia furfuracea (L.) Zopf (Parmeliaceae) is a common epiphytic lichen in the conifer-hardwood forest of Anatolia. This species is used in traditional medicine in Turkey as a treatment for wounds, eczema and hemorrhoids.

Aim of the study

The present study was designed to investigate the active compounds from Pseudevernia. furfuracea, and the isolation studies yielded atraric acid ( Aslan et al., 2006) as the major compound and a mixture of methyl hematommate ( Baumann, 1960) and methyl chlorohematommate ( Bay?r et al., 2006). Furthermore, methanolic extract from thalli of Pseudevernia. furfuracea and its fractions and isolates ( 1, 2 and 3) were investigated for in vitro antimicrobial and antioxidant activities, and in vivo antinociceptive, anti-inflammatory and wound healing activities.

Material and methods

Antimicrobial activities of the samples were determined by using the disc diffusion technique. 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay was used as a rapid TLC screening method to evaluate the antioxidant activity of Pseudevernia. furfuracea. The thiobarbituric acid (TBA) test was used to assess the efficacy of the extracts in protecting liposomes from lipid peroxidation. In vivo inhibitory effect of the extracts on the carrageenan-induced hind paw edema model in mice was studied for the assessment of anti-inflammatory activity. p-Benzoquinone-induced abdominal constriction test was used to explore the antinociceptive effects of the extracts. Moreover, the wound healing potential of the plant extracts that were evaluated by using in vivo incision and excision wound models on rats and mice, were comparatively assessed with a reference ointment Madecassol^®.

Results

Significant antimicrobial activities were observed against Gram (+) microorganisms and Candida krusei and Candida. dubliniensis in dichloromethane (DCM) and ethyl acetate (EtOAc) extracts and isolates. The methanol (MeOH), DCM and EtOAc extracts of the lichen were found to possess moderate inhibitory activity on lipid peroxidation. Methanolic extract of the lichen was found to possess significant inhibitory activity on the carrageenan-induced hind paw edema model in mice whereas the other fractions did not show any activity. While DCM and EtOAc extracts and fractions showed notable anti-inflammatory activity on carrageenan-induced hind paw edema model without inducing any apparent acute toxicity or gastric damage. Moreover, topical application of the ointment prepared with MeOH extract and EtOAc fraction onto the incised wounds exerted remarkable wound healing activity.

Conclusion

The results of these experimental studies exhibited that nonpolar fractions of Pseudevernia. furfuracea have significant antimicrobial activity against especially Candida species and polar fractions (especially MeOH) display antioxidant, anti-inflammatory, antinociceptive and wound healing activities.     

A novel wound healing ointment: a formulation of Hypericum perforatum oil and sage and oregano essential oils based on traditional Turkish knowledge

Aim of the study

Hypericum perforatum L. (Hypericaceae), olive oil (Oleaceae), Origanum Tourn ex L. and Salvia L. species (Lamiaceae) are used against inflammatory disorders and for healing of skin wounds in traditional Turkish medicine. A new ointment formulation was developed to provide more efficient wound healing activity. The content of the formulation was as follows; olive oil extract of flowering aerial parts of Hypericum perforatum L., olive oil, an equivalent mixture of Origanum majorana L. and Origanum minutiflorum Schwrd. et Davis essential oils (Origani aetheroleum), Salvia triloba L. essential oil. The aim of the present study is to assess the wound healing potential of this new formulation by using in vivo and in vitro models as well as histopathological methods.

Materials and methods

For the evaluation of wound healing potential of this formulation (HPP crème mit Rotöl ointment) in vivo wound healing experimental models were employed on rats and mice and the efficiency was comparatively assessed against a reference ointment Madecassol^®. Tissue sections were also evaluated histopathologically. Furthermore, the wound healing activity of each component was also investigated individually to determine the improvement in the healing capacity of the formulation.

Results

The ointments of HPP crème mit Rotöl and Hypericum perforatum L. demonstrated the highest activities on both wound models when compared to reference ointment Madecassol^®, while the other ingredients did not show any remarkable wound healing effect. However, the efficacy of the formulation was remarkably higher than the Hypericum L. ointment alone which was also confirmed by histopathological evaluation. On the other hand, the formulation did not reduce elastase activity in vitro, but inhibited the collagenase activity. HPP crème mit Rotöl also exerted bactericidal and candicidal activities.

Conclusions

The experimental studies revealed that HPP crème mit Rotöl formulation displays remarkable wound healing activity. To be acting on the different stages of wound healing process could be considered as a beneficial effect of the formulation for the treatment of wounds.     

Determination of the wound healing effect of Calendula extracts using the scratch assay with 3T3 fibroblasts

Pharmacological relevance

Presentation of the scratch assay as a convenient and inexpensive in vitro tool to gain first insights in the wound healing potential of plant extracts and natural compounds.

Aim of the study

The present study deals with the optimization of the scratch assay which can be used as an in vitro model for quantification of fibroblast migration to and proliferation into the wounded area. It is suitable for the first evaluation of the wound re-epithelialization potential of crude herbal extracts, isolated compounds and pharmaceutical preparations. As a proof of concept three preparations from traditional medicinal plants were investigated.

Materials and methods

Swiss 3T3 albino mouse fibroblasts were used in monolayers and platelet derived growth factor as positive control. Hexane and ethanolic extracts from Calendula officinalis and Matricaria recutita, Hypericum oil as well as the triterpenoids faradiol myristate and palmitate were studied. To differentiate between proliferation and migration antimitotic mitomycin C was added.

Results

Both extracts of Calendula officinalis stimulated proliferation and migration of fibroblasts at low concentrations, e.g. 10 μg/ml enhanced cell numbers by 64.35% and 70.53%, respectively. Inhibition of proliferation showed that this effect is mainly due to stimulation of migration. Faradiol myristate and palmitate gave comparable stimulation rates at an almost 50 μg/ml concentration, indicating that they contribute partially, but not most significantly to the wound healing effects of Calendula preparations. Extracts from Matricaria recutita were only moderately active. Hypericum oil was cytotoxic at concentrations higher than 0.5 μg/ml.

Conclusions

The scratch assay in the present form can be used as a promising scientific approach and platform to differentiate between plant extracts known for their wound healing and their anti-inflammatory properties.     

Inhibitory effect of St. John׳s Wort oil macerates on TNFα-induced NF-κB activation and their fatty acid composition

Ethnopharmacological relevance

The oil macerates of Hypericum perforatum L. (St. John?s Wort=SJW) have a long history of medicinal use and SJW has been used in traditional medicine both orally and topically for centuries worldwide mainly for wound healing, ulcer and inflammation.

Materials and methods

We analyzed the fatty acid composition of 10 traditionally (home-made) and 13 commercially (ready-made) prepared SJW oil macerates by GC–MS. The acid, peroxide, iodine, saponification values, and the unsaponifiable matters of the samples were determined according to the European Pharmacopoeia. We also explored potential mechanism of wound healing effect of the samples, i.e. TNFα-induced NF-κB activation.

Results

The most home-made oil samples contained oleic acid predominantly and complied with the requirements set for olive oil by European Pharmacopoeia. However, majority of the ready-made samples appeared to have adulteration with some other oils. Moderate NF-κB inhibitory effects have been observed with some of the oil samples.

Conclusion

This study sheds light on the fact that application of the proper traditional method to prepare olive oil macerates from Hypericum perforatum is able to get bioactive constituents in the oil. Besides, inhibition of TNFα-induced NF-κB activation appears to be a potential mechanism for topical wound healing activity of SJW oil macerates.     

Hypericum grandifolium Choisy: A species native to Macaronesian Region with antidepressant effect

Aim of the study

Various species of Hypericum genus have been used in the Canary Islands as sedative, diuretic, vermifuge, wound healing, antihysteric and antidepressant agent. Studies have shown that methanol extract of Hypericum grandifolium Choisy is active in tetrabenazine-induced ptosis and forced swimming tests. In the current study, the aqueous, butanol and chloroform fractions obtained from the methanol extract as well as three sub-fractions derived from the chloroform fraction were evaluated for their central nervous effects in mice, particularly their antidepressant activity.

Materials and methods

The central nervous effect of different fractions and sub-fractions of Hypericum grandifolium was evaluated in mice using various behavioural models including locomotor and muscle relaxant activity, forced swimming test, effect on normal body temperature, barbiturate-induced sleep, tetrabenazine-induced syndrome and 5-hydroxytryptohan-induced head twitches and syndrome.

Results

We found that the butanol and chloroform fractions and all sub-fractions showed an antidepressant effect in the forced swimming test, the chloroform fraction being the most active. They produced no effects or only a slight depression of locomotor activity. Chloroform fraction significantly increased the pentobarbital-induced sleeping time, produced a slight but significant hypothermia and antagonized tetrabenazine-induced ptosis, whereas the butanol fraction produced a slight potentiation of 5-HTP-induced head twitches and syndrome.

Conclusions

The present results, together with previous pharmacological and phytochemical data, indicated that Hypericum grandifolium possess antidepressant-like effects in mice and that different constituents, such as the flavonoids and the benzophenone derivatives, could be responsible at least in part for the antidepressant effects observed for this species.     

Evaluation of the wound healing property of Boesenbergia longiflora rhizomes

Ethnopharmacological relevance

The rhizomes of Boesenbergia longiflora (Wall.) Kuntze (Zingiberaceae) have been traditionally used for treatment of inflammatory bowel disease, ulcerative colitis, aphthous ulcer and abscess by decoction with alcohol.

Aim of the study

The rhizomes of Boesenbergia longiflora were carried out to investigate for anti-inflammatory and wound healing activities in order to support the traditional use.

Material and methods

The ethanolic extract of Boesenbergia longiflora and its fractions were tested using relevant in vitro anti-inflammatory and wound healing assays. For the in vitro studies, murine macrophage RAW264.7 cells and mouse fibroblast L929 cells were assessed for anti-inflammatory and fibroblast stimulatory activities, respectively. In vivo anti-inflammatory activity was determined by carrageenan-induced rat paw edema model as well as acute toxicity estimated by the up-and-down method in mice.

Results

The present study has demonstrated that the ethanolic extract of Boesenbergia longiflora rhizomes possesses a potent anti-inflammatory and wound healing activities. Among the isolated fractions, the CHCl₃ fraction showed potent anti-inflammatory effect through nitric oxide inhibitory activity (IC₅₀=5.5 μg/ml) and reduction of carrageenan-induced rat paw edema (ED₅₀=222.7 mg/kg), whereas this fraction exhibited wound healing property via fibroblast migration on both day 1 (77.3%) and day 2 (100%) as well as enhanced collagen production (187.5 μg/ml) at concentration of 3 μg/ml, compared to that of the controls, 39.4% for fibroblast and 60.8 μg/ml for collagen, respectively. The anti-inflammatory mechanism of the CHCl₃ fraction is found to suppress the iNOS and COX-2 mRNA expression.

Conclusion

The scientific investigation of wound healing activity of Boesenbergia longiflora rhizomes support the Thai traditional uses for treatment of inflammatory bowel disease, ulcerative colitis, aphthous ulcer and abscess. The EtOH extract and CHCl₃ fraction exert potential wound healing property through NO inhibition, anti-oxidant effect and stimulation of fibroblast migration and collagen production. The phytochemical screening revealed that the CHCl₃ fraction of Boesenbergia longiflora rhizomes contains diarylheptanoids, flavonoids and terpenes. The isolation of the compounds responsible for the wound healing effect is now in progress.     

Anti-ulcerogenic activity and isolation of the active principles from Sambucus ebulus L. leaves

Ethnopharmacological relevance

Sambucus ebulus L. has a very prominent place in Turkish folk medicine. Leaves of the plant are practiced externally to relieve rheumatic pain, to treat abscess, for wound healing and internally against hemorrhoids and stomachache. In a previous work, aqueous extract of the leaves was shown to possess potent antiulcerogenic activity on water immersion and immobilization-induced stress ulcer model in rats. This study aims to investigate the antiulcerogenic activity profile of the plant on various in vivo peptic ulcer models and gastric biochemical parameters and through bioassay-guided processing to isolate the active constituent (s) and to elucidate its structure.

Materials and methods

Among the subextracts obtained by successive solvent extractions from the MeOH extract of the leaves, the butanol subextract exerted significant antiulcerogenic activity against water-immersion and immobilization-induced stress ulcer model in rats as the bioassay model. This subextract was then subjected to successive chemical separation techniques (precipitation, column chromatography based on ion-exchange, silica gel and sephadex) and the activity of each fraction/subfraction was tested using the same bioassay model. After determination of active principles, further studies were performed on the active subextract by using various in vivo test models (ethanol-, serotonin-, pyloric ligation-induced ulcerogenesis) in rats as well as biochemical methods for the evaluation of antiulcerogenic potential.

Results

Bioassay-guided fractionation procedures yielded two flavonol glycosides as the active antiulcerogenic principles. The structures of these compounds were elucidated as isorhamnetin-3-O-monoglycoside and quercetin-3-O-monoglycoside by using ¹H, ¹³C-NMR, and FAB-MS techniques.

Conclusion

This study has proven the folkloric use of Sambucus ebulus leaves for the treatment of gastric ailments in Turkish folk medicine. The antiulcerogenic activity of the two flavonol glycosides isolated in the present study was not previously reported elsewhere.     

Wound healing activity of latex of Euphorbia caducifolia

Ethanopharmacological relevance

Euphorbia caducifolia is a Euphorbiaceae species native to Thar Desert of India, where latex of E. caducifolia (ECL) is used by the local inhabitants for treatment of bleeding wound, cutaneous eruption and other skin diseases. Looking to the medicinal importance of the plant, it was considered worthwhile to scientifically evaluate the ECL for the wound healing activity.

Material and methods

In vitro methods, clotting of platelet free plasma and angiogenesis in chick chorioallantoic membrane (CCM) were used for studying effect of ECL on clotting and angiogenesis. Excision and incision wounds model were used to study effect of ECL on wound contraction, tensile strength and hydroxyproline and DNA content.

Results

ECL treatment stimulates the blood vessel formation in CCM, at the all dose levels. 56.77, 74.48 and 78.09% increase in area of angiogenesis was observed in ECL 2.5, 5.0 and 10 mg/ml treated groups respectively. The excised skin of ECL 5.0 and 10 mg/ml treated animals found to have higher content of hydroxyproline and DNA, when compared with control group. An acceleration of the wound closure was observed with an enhanced number of fibroblasts and collagen content in ECL treated mice.

Conclusion

Result of in vitro and in vivo experiments indicate that ECL exerts significant wound healing activity, demonstrating its relevant therapeutic potential.     

Wound healing effect of Flabellaria paniculata leaf extracts

Aim of the study

This study evaluated wound healing activity of the chloroform and aqueous fractions of the methanolic extract of Flabellaria paniculata leaf on infected wounds in rats as a follow up to an initial study of the crude extract.

Materials and methods

Wounds were inflicted on Wistar rats using excision model. Local infection was introduced into rat abdominal wounds with Staphylococcus aureus and Pseudomonas aeruginosa using a 10⁸ cells/ml inoculum. Rate of wound healing was assessed by contraction and period of epithelization.

Results

Chloroform fraction exhibited significant wound healing potency (p < 0.05) as compared with controls. The test drug achieved 100% wound contraction on day 14 in non-infected group, on day 16 in Staphylococcus aureus infected group and on day 18 in Pseudomonas aeruginosa group.

Conclusions

Chloroform extract of Flabellaria paniculata proves to be a potential anti-infective and wound healing agent. Its in vitro antibacterial and in vivo wound healing activities are in good agreement with the local medicinal use of the plant for skin diseases and sores.     

Evaluation of in vivo wound healing activity of methanol extract of Achyranthes aspera L

Ethnopharmacological relevance

The leaves of Achyranthes aspera L. (Amarenthacea) has been used traditionally for the treatment of wound in various parts of Ethiopia. However, the plant has not been explored scientifically for its wound healing activity. Therefore, this study was designed to investigate the wound healing activity of methanol extract of Achyranthes aspera L. leaves in rats.

Materials and methods

Incision and excision wounds were inflicted on albino rats of either sex, under diethyl ether anesthesia. Group I served as positive control and was treated with 1% silver sulphadiazine, group II, III, IV treated with simple ointment containing 2.5%, 5% and 10% (w/w) methanol extract of the leaves of Achyranthes aspera L., respectively, whereas group V served as negative control and was treated with simple ointment. All the animals were treated topically once a day. Wound healing potential was assessed with excision and incision wound model. Excision wound model was used to assess the change in percentage contraction of wound, epithelization time, DNA content and histological features whereas rats inflicted with the incision wounds were used to determine breaking strength.

Results

Based on the results of percentage wound contraction, the DNA content and epithelization time, all groups of rats treated with methanol extract of the leaves of Achyranthes aspera L. showed significant (p<0.05) wound healing activity compared to group of rats treated with simple ointment (negative control) group. The difference in breaking strength was, however, significant (p<0.05) only for the 5% and 10% methanol extract of Achyranthes aspera (w/w) ointment treated groups. Histological evaluation showed well organized epidermal layer, increased number of fibrocytes, remarkable degree of neovascularization and epithelization which was comparable to the standard on the 21st day after treatment; especially in the 5% and 10% (w/w) extract treated group.

Conclusion

The present study provides a scientific rationale for the traditional use of the leaf extracts of Achyranthes aspera L. in the treatment of wound.     

Comparative assessment of dermal wound healing potentials of various Trifolium L. extracts and determination of their isoflavone contents as potential active ingredients

Ethnopharmacological relevance

Trifolium species are used in Turkish folk medicine as a wound healing agent, expectorant, antiseptic, sedative and to alleviate pain in rheumatism. In the present study, the aqueous methanolic extracts (80%) of 13 Trifolium species (Trifolium ambigum, Trifolium arvense var. arvense, Trifolium campestre, Trifolium canescens, Trifolium hybridum var. anatolicum, Trifolium hybridum var. hybridum, Trifolium pannonicum, Trifolium pratense var. pratense, Trifolium purpureum var. purpureum, Trifolium repens var. repens, Trifolium resupinatum var. microcephalum, Trifolium spadiceum and Trifolium trichocephalum) collected from different regions of Anatolia were evaluated for their in vivo wound healing effects.

Materials and methods

In vivo wound healing activities of the plant aqueous methanolic extracts were evaluated by linear incision and circular excision wound models subsequent to histopathological analysis. Active constituents were determined by a validated high performance liquid chromatographic method. Precision of the method was performed by the evaluation of intra-day and inter-day variations of the each standard at limits of quantification (LOQ) levels.

Results

The aqueous methanolic extracts of Trifolium canescens and Trifolium pretense var. pratense possessed better wound healing activity compared to the other extracts and control groups. The animal groups treated with the Trifolium canescens extract demonstrated increased contraction (48.96%) on excision and a significant increase in wound tensile strength (35.6%) on incision models. The main compounds were detected as genistein and biochanin A for Trifolium canescens.

Conclusion

The results of the present study revealed the wound healing potential of Trifolium canescens. This might be due to the combined effect of the isoflavones genistein, formononetin, daidzein, and biochanin A present in the extract.     

In vivo antioxidative property, antimicrobial and wound healing activity of flower extracts of Pyrostegia venusta (Ker Gawl) Miers

Ethnopharmacological relevance

Pyrostegia venusta (Ker Gawl) Miers. (Bignoniaceae), has been traditionally used as a remedy for treating white patches and infections on the skin (leukoderma, vitiligo).

Aim of the study

To investigate wound healing and antimicrobial activity of flower extract of Pyrostegia venusta, including in vivo antioxidant activity.

Materials and methods

Methanolic extracts of Pyrostegia venusta flowers were studied for wound healing efficiency along with its effect on pro-inflammatory and anti-inflammatory cytokines was assessed using excision and incision model of wound repair in Wistar rats. Healing was assessed by the rate of wound contraction, tensile strength, breaking strength, hydroxyproline and hexosamine content. Antimicrobial activity of the flower extract against twelve microorganisms was also assessed. In vivo antioxidant activity was performed to understand the mechanism of wound healing potency.

Results

The results indicated that Pyrostegia venusta extract has potent wound healing capacity as evident from the wound contraction and increased tensile strength. Hydroxyproline and hexosamine expression were also correlative with the healing pattern observed. Pyrostegia venusta extract exhibited moderate antimicrobial activity against the organisms: Bacillus subtilis, Staphylococcus epidermidis, Staphylococcus pyogenes, Staphylococcus aureus, Escherichia coli, Micrococcus luteus, Enterobacter aerogenes, Salmonella typhi, Pseudomonas aeruginosa, Candida albicans, Aspergillus niger and Candida tropicana. During early wound healing phase TNF-α and IL-6 level were found to be up regulated by Pyrostegia venusta treatment.

Conclusion

Increased wound contraction and tensile strength, augmented hydroxyproline and hexosamine content along with antioxidative activity and moderate antimicrobial activity support the early wound healing exhibited by Pyrostegia venusta flower extract. Induction in cytokine production may be one of the mechanisms involved in accelerating the wound healing by Pyrostegia venusta extract. Results suggest that Pyrostegia venusta may be useful in the tropical management of wound healing.     

Eleutherine indica L. accelerates in vivo cutaneous wound healing by stimulating Smad-mediated collagen production

Ethnopharmacological relevance

Eleutherine indica L. has been used for healing of wound, painful and irregular menstruation, dysentery and lesions, and topically used as antiseptic and antimicrobial agent in folk medicine. In the present study, methanolic extracts of Eleutherine indica was subjected to scientific investigation for in-vivo cutaneous wound healing in wistar rat.

Materials and methods

In-vivo wound healing activity of Eleutherine indica was evaluated by using circular excision experimental models, followed by histopathological and western blot analysis. The healing potential was comparatively assessed with a reference gentamicin sulfate hydrogel (0.01% w/w). Wound contraction measurement, hydroxyproline estimation and western blot for COL3A1, bFGF, Smad-2, -3, -4, and -7 were performed.

Results

The methanolic extract of Eleutherine indica showed accelerated wound healing activity as evidenced by fast wound contraction rate and higher hydroxyproline content of granulation tissue. Western blot revealed the Smad-mediated collagen production promoting property of Eleutherine indica methanolic extract. Histopathological examinations also supported the experimental findings.

Conclusion

The study revealed that Eleutherine indica promotes wound healing by augmenting Smad-mediated collagen production in wound granulation tissue.