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.     

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.     

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.     

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.     

Wound healing potential of methanolic extract of Trichosanthes dioica Roxb (fruits) in rats

Aim of the study

The present study provides a scientific evaluation for the wound healing potential of methanolic (MeOH) extract of TDR fruits.

Materials and methods

Excision and incision wounds were inflicted upon three groups of six rats each. Group I was assigned as control (ointment base), Group II was treated with standard silver sulfadiazine (0.01%) cream. Group III was treated with 5% MeOH extract ointment. The parameters observed were percentage of wound contraction, epithelialization period, hydroxyproline content, tensile strength including histopathological studies.

Results

It was noted that the effect produced by the extract ointment showed significant (P < 0.01) healing in both the wound models when compared with control group. All parameters such as wound contraction, epithelialization period, hydroxyproline content, tensile strength and histopathological studies showed significant changes when compared to control.

Conclusion

The result shows that TDR extract ointment demonstrates wound healing potential in both excision and incision models.     

Wound healing potential of Sambucus ebulus L. leaves and isolation of an active component,quercetin 3-O-glucoside

Ethnopharmacological relevance

The leaves of Sambucus ebulus L. are used in Turkish folk medicine for treatment of high fever, rheumatic pains, snake bites and wounds.

Aim of the study

Four different solvent extracts by using n-hexane, diethyl ether, ethyl acetate and methanol were prepared from the leaves of Sambucus ebulus in order to investigate the claimed wound healing activity in vivo and isolation of active component(s) from the active extract through the bioassay-guided fractionation procedures.

Materials and methods

Linear incision and circular excision wound models on rats and mice were employed. The methanol extract was fractionated by successive chromatographic techniques. Wound healing activity of each fraction was investigated following the bioassay-guided fractionation procedures. Moreover, the tissue samples were examined histopathologically.

Results

Significant wound healing activity was observed for the ointment prepared with methanol extract at 1% concentration. The methanol extract treated groups of animals showed 84.3% contraction in circular excision model, which was close to contraction value of the reference drug Madecassol^® (100%). On the other hand, a significant increase (43.7%) in the wound tensile strength was determined with the same extract on incision wound model. Subfractions showed significant but reduced wound healing activity on both in vivo wound models. A flavonoid derivative “quercetin 3-O-glucoside” was isolated and determined as one of the active component of active final subfraction. The results of histopathological examination supported the outcome of linear incision and circular excision wound models.

Conclusion

The experimental data revealed that the methanolic extract of Sambucus ebulus leaves displayed remarkable wound healing activity.     

Investigations on the in vivo wound healing potential of Hypericum perforatum L.

Ethnopharmacological relevance

Olive oil extract of the flowering aerial parts of Hypericum perforatum L. (Hypericaceae) is a popular folk remedy for the treatment of wounds in Turkey.

Aim of the study

In order to prove the claimed utilization of the plant, the effects of the extracts and the fractions were investigated by using bioassay-guided procedures. For the wound healing activity assessment, in vivo excision and incision wound models were applied. For the anti-inflammatory activity, an in vivo model, based on the inhibition of acetic acid-induced increase in capillary permeability was used as well. Moreover, a parallel study was run on Hypericum scabrum L., which is a widespread species of the gender but not known as a folk remedy for wound healing, to provide a preliminary data to compare and emphasize the selection of correct plant species.

Results

Initial investigations proved that the olive oil extract of Hypericum perforatum has a significant wound healing effect on excision (5.1–82.6% inhibition) and circular incision (20.2–100.0% inhibition) wound models. In order to determine the active wound healing ingredient(s), aerial parts of the plant was extracted with ethanol, noteworthy wound healing activity profile was observed with the wound models; between 18.3% and 95.6% in excision model and from 13.9% to 100.0% inhibitions in incision model were determined. The ethanolic extract was then submitted to successive solvent extractions with n-hexane, chloroform and ethyl acetate (EtOAc). Each solvent extract was also applied on the same wound models, consequently, EtOAc subextract was found to be the most active one by inhibiting wounds between 17.9% and 100.0% in excision model, subsequently between 9.4% and 100.0% in incision model. However, all subfractions obtained from the EtOAc subextract using Sephadex LH-20 column chromatography showed wound healing activity not more than the whole EtOAc subextract, which revealed that a possible synergistic activity that might be questioned. Among the active Sephadex fractions, Fr. A further yielded hyperoside, isoquercitrin, rutin and (−)-epicatechin and Fr. B yielded hypericin as the major components. Moreover, a dose-dependent anti-inflammatory activity was found for the ethanol extract, EtOAc subextract and Sephadex fractions of Hypericum perforatum. These results suggest that anti-inflammatory activity of the active fractions might have a contributory role in the wound healing effect of the plant.

Conclusion

Results of the present study have proved that aerial parts of Hypericum perforatum possess remarkable wound healing and anti-inflammatory activities supporting the folkloric assertion of the plant in Turkish folk medicine. Flavonoids [hyperoside, isoquercitrin, rutin and (−)-epicatechin] and naphthoquinones (hypericins) were found as the active components of Hypericum perforatum. On the other hand, ethanol extract of Hypericum scabrum showed neither remarkable wound healing nor anti-inflammatory activity demonstrating the importance of correct plant species selection in therapeutic applications.     

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.     

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.     

Evaluation of the wound healing activity of methanol extract of Pedilanthus tithymaloides (L.) Poit leaf and its isolated active constituents in topical formulation

Ethnopharmacological relevance

Pedilanthus tithymaloides leaves are widely used in Indian medicine to heal wounds, burn, mouth ulcers. However, systematic evaluation of these activities is lacking. Thus, the present study aimed to assesses the wound healing activity of Pedilanthus leaves and its isolated constituents in topical ointment formulation.

Materials and Methods

Bioassay-guided chromatographic fractionation of the methanol extract of leaves resulted in the isolation of 2-(3,4-dihydroxy-phenyl)-5,7-dihydroxy-chromen-4-one and 1, 2-tetradecanediol, 1-(hydrogen sulfate), sodium salt. The ointment formulation of methanol extract (2.5%, 5% w/w) and isolated compounds (0.25% w/w) was prepared and evaluated on excision, incision and dead space wound models in rats. The effects of formulations on wound healing were assessed by the rate of wound closure, period of epithelialization, tensile strength, granulation tissue weight, hydroxyproline content and histopathology.

Results

Significant wound healing activity was observed with methanol extract and isolated constituents. Topical application of isolated compound ointments caused faster epithelialization, significant wound contraction (95.41%), and better tensile strength (565.33 g) on 16 post-wounding day, while 5% extract showed wound epithelialization with 95.55% contraction on 18th post-wounding day, better than the control group (76.39% on 22 day). The tensile strength of incision wound was significantly increased in extract and compound treated animals. Moreover, in dead space model the extract significantly increased granuloma tissue weight, tensile strength and hydroxyproline content. The tissue histology of ointment treated groups showed complete epithelialization with increased collagenation, compared to the povidone–iodine group.

Conclusions

The results validated the traditional use of Pedilanthus tithymaloides for cutaneous wound management.     

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.     

In vivo topical anti-inflammatory and wound healing activities of the fixed oil of Caryocar coriaceum Wittm. seeds

Ethnopharmacological relevance

Caryocar coriaceum Wittm. (Caryocaraceae), popularly known as pequi, is important source of food and used in the folk medicine as wound healing, anti-inflammatory and for the treatment of diseases of the respiratory tract, rheumatic and muscular pains and gastric ulcer.

Aim of the study

To evaluate the effects of fixed oil of Caryocar coriaceum (FOCC) on the topical inflammation and cutaneous wound healing.

Materials and methods

FOCC was purchased commercially and used at 6%, 12%, 25%, 50% and 100% on model of ear edema induced by xylene and ointments formulations at 6% and 12% on wound excision model in mice. Skin lesions were evaluated by planimetry and histological analysis.

Results

FOCC was capable of reducing inflammation in a dose-dependent fashion. FOCC in nature (100%) inhibited the ear edema in 38.01% and 39.20% at time 15 min and 1 h, respectively, after induction of inflammation, when compared to the positive control. The topical administration of FOCC ointment 12% showed a significantly reduction of unhealed wound area, with increased percentage of wound contraction (96.54%) on day 7 when compared to the other groups. The results of histological evaluation supported the outcome of excision wound model.

Conclusion

Caryocar coriaceum inhibits the topical inflammation and accelerate cutaneous wound repair.     

Topical application of serine proteases from Wrightia tinctoria R. Br. (Apocyanaceae) latex augments healing of experimentally induced excision wound in mice

Ethnopharmacological relevance

Wrightia tinctoria R. Br. (Apocyanaceae) is a folk medicinal plant known to have immunomodulatory, anti-inflammatory and antihemorrhagic potential. Wrightia tinctoria latex is used for treatment of various clinical conditions including psoriasis, blisters, mouth ulcers, and extensively for topical application on fresh wounds to promote accelerated healing.

Aims of the study

To investigate the wound healing potential of Wrightia tinctoria latex proteases using a mouse model.

Materials and methods

Proteolytic activity of Wrightia tinctoria latex proteases (WTLP) was determined on various substrates (casein, gelatin and collagen (type-I and IV)). The thermal stability and the class of proteases present in WTLP were determined using heat treatment and specific protease inhibitors, respectively. Excision wound model in mice was used to evaluate the healing potential of WTLP application (twice daily, 10 mg/kg). Neosporin, a standard drug, was used for comparison. The progression of healing was monitored using physical (wound contraction), biochemical (collagen content, catalase and MMP activity) and histological examinations.

Results

WTLP contains thermostable serine proteases, which are completely inhibited by PMSF. WTLP showed strong caseinolytic, gelatinolytic and collagenolytic activity. The excision wound healing rate upon WTLP treatment was significantly higher than (>2-fold) the control group (49% vs. 18%, ^??p<0.01) on day 3 and throughout the study. PMSF pre-treated and heat denatured WTLP failed to promote wound healing. In addition, serial biochemical analysis of the granulation tissue demonstrated 1.5-fold more (2444±100 vs. 1579±121 µg/100 mg tissue) hydroxyproline content and 5.6-fold higher catalase activity (16.7±1.3 vs. 3±0.3 units/mg) compared to controls. Further, the enhanced collagen content and matrix metalloproteinase activity correlated with wound contraction rate following WTLP and Neosporin treatment. Histological analysis on day 9 confirmed complete epithelialization, re-establishment of skin structure and accelerated wound healing following WTLP treatment.

Conclusions

The thermostable serine proteases of Wrightia tinctoria latex are directly involved in the wound healing process. Our findings provide a biochemical basis for the role of WTLP in the enhancement of wound healing. The study supports traditional topical application of Wrightia tinctoria latex on fresh wounds to promote accelerated healing.     

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

Ethnopharmacological Relevance

Salvia L. species are widely used against wounds and skin infections in Turkish folk medicine.

Aim of the study

The aim of the present study is to evaluate wound healing activity of the ethanol (EtOH) extracts of Salvia cryptantha and Salvia cyanescens.

Materials and methods

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.

Results and Conclusions

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.     

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.     

Topical effects of frog “Rana ridibunda” skin secretions on wound healing and reduction of wound microbial load

Ethnopharmacological relevance

Study of the interrelationships between human and the animals in their environment has always been a subject of interest and caused discoveries of the animal applications in medicine. From the latest century, these remedies called back in traditional medicine of Vietnam and South America and frog skin was used as a biological dressing and had good effects in healing wounds. Also, frog skin secretions have wound healing properties and reduce inflammation. In this study we applied these secretions in the form of ointment to investigate their healing activities.

Materials and methods

Skin secretions were extracted from Rana ridibunda to evaluate their effects on wound healing in mice. Secretion used as raw extract (RE) and ultrafiltrated extract, using a membrane with cutoff 10 kDa as under 10 kDa (U10E), was administrated as ointment every 48 h on wound site. Control group was left without any treatment and also there was other group treated with ointment (O group) alone. On 2, 4 and 6 days post injury, animals were euthanized and images were taken for wound closure evaluation. Then wound locations were removed for histological assays. Also wound microbial load was examined. Observational parameters including wound closure and wound microbiology in experimental groups compared with the control and O groups have been studied.

Results

The results showed U10E group has better effects than RE, O and control groups. Histological parameters, including numbers of inflammatory and fibroblast cells and amount of collagen fibers, neovascularization, as well, represented greater degree of wound healing in U10E group compared with RE, O, and control groups.

Conclusions

Our results showed that frog skin secretions were significantly effective in promoting wound healing process. The U10E extract from the frog R. ridibunda possesses a potent accelerating wound healing effect that promises good potential for clinical application in wound care. Further studies will be required to characterize special molecules encompassing healing properties.     

The potential role of female flowers inflorescence of Typha domingensis Pers. in wound management

Aim of the study

Female flowers inflorescence of Typha species including Typha domingensis Pers. are used externally for burns and wound healing in Turkish folk medicine. In order to verify the folkloric assertion, the female and male flowers inflorescences were individually submitted to in vivo wound models.

Material and methods

Ointment formulations prepared directly either from the male or female flowers inflorescences of Typha domingensis in 5% and 10% concentrations were submitted to activity testing. After that, female flowers inflorescence was further submitted to successive extractions with solvents in increasing polarity; i.e., n-hexane, chloroform, methanol and water and the wound healing activity of each extract was investigated. The linear incision and circular excision wound models were used for the evaluation of the healing potential of the test materials in rats and mice. Tissue sections were also evaluated by histopathological techniques.

Results

Remarkable wound healing activity was observed only for the female flowers inflorescence at 5% concentration in ointment base and its methanolic and aqueous extracts. The wound healing effect was found comparable to that of reference ointment Madecassol^®. The results of histopathological evaluation supported the outcome of both linear incision and circular excision wound models.

Conclusion

The experimental study revealed that the female flowers inflorescence of Typha domingensis displayed notable wound healing activity in mice and rats, at the models tested.     

The traditional Hungarian medicinal plant Centaurea sadleriana Janka accelerates wound healing in rats

Ethnopharmacological relevance

The aerial parts of Centaurea sadleriana Janka, a species native to Hungary, have been used for the healing of wounds of livestock in Hungarian folk medicine. This is the first report of the ethnomedicinal use of this plant.

Aim of the study

This study was aimed at investigating the wound-healing efficiency of different extracts of Centaurea sadleriana.

Materials and methods

Experimental wounds inflicted on healthy rats by means of a branding iron were treated topically with different extracts and fractions of extracts of the aerial parts of Centaurea sadleriana. To assess the effectiveness of treatment, an absolute control (no treatment), a vehicle control (Carbomer gel) and a positive control group (1% salicylic acid in Carbomer gel) were applied.

Results

The n-hexane fraction of the methanol extract significantly accelerated the wound-healing process. This effect was rather similar to that of the positive control gel. Other fractions exhibited more moderate activities.

Conclusions

The apolar fraction of the methanol extract of Centaurea sadleriana facilitated wound healing significantly, corroborating the folk medicinal use of this plant.