Wound repair and anti-inflammatory potential of essential oils from cones of Pinaceae: preclinical experimental research in animal models

Ethnopharmacological relevance

Ethnobotanical surveys revealed that Abies bornmulleriana, Abies cilicica, Abies nordmanniana and Cedrus libani have been used to promote wound healing in Turkish folk medicine. Four different fir species (Abies cilicica subsp. cilicica, Abies nordmanniana subsp. bornmulleriana, Abies nordmanniana subsp. equi-trojani, and Abies nordmanniana subsp. nordmanniana), Cedrus libani and Picea orientalis were assessed for their in vivo wound healing and anti-inflammatory activities.

Materials and methods

The essential oils from six different coniferous cones were used. In vivo wound healing activity of the plants was evaluated by linear incision and circular excision experimental wound models subsequently histopathological analysis. The healing potential was comparatively assessed with a reference ointment Madecassol^®, which contains 1% extract of Centella asiatica. Additionally acetic acid-induced capillary permeability test was used for the oils’ anti-inflammatory activity.

Results

The essential oils from Cedrus libani and Abies cilicica subsp. cilicica demonstrated the highest activities on the both wound models. Moreover, the oil from Abies nordmanniana subsp. bornmulleriana was found generally highly effective. On the other hand, the rest of the species did not show any remarkable wound healing effect. Results of the present study support the continued and expanded utilization of these plant species employed in Turkish folk medicine.

Conclusion

The experimental study revealed that Cedrus libani and Abies cilicica subsp. cilicica display remarkable wound healing and anti-inflammatory activities.     

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 and anti-inflammatory properties of Ranunculus pedatus and Ranunculus constantinapolitanus: a comparative study

Ethnopharmacological relevance

In Turkish folk medicine Ranunculus species are used for wound healing and for the treatment of rheumatism. The present study was conducted to evaluate in vivo wound healing and anti-inflammatory properties of Ranunculus pedatus and Ranunculus constantinapolitanus.

Material and methods

In vivo wound healing activity of the extracts prepared from Ranunculus pedatus and Ranunculus constantinapolitanus was evaluated by linear incision and circular excision wound models. Hydroxyproline content of the treated tissues was also assessed. We also studied the anti-inflammatory activity using Whittle method with some modifications.

Results

Methanolic extract of Ranunculus pedatus showed significant wound healing effect both in incision (31.4%) and excision (55.74%) wound models. Methanolic extract of both Ranunculus pedatus and Ranunculus constantinapolitanus demonstrated anti-inflammatory activity with the inhibition value of 26.2% and 23.3% respectively, at the dose of 100 mg/kg. Hydroxyproline content of the tissues treated with the methanolic and aqueous extracts of Ranunculus pedatus and methanolic extract of Ranunculus constantinapolitanus were found to be significantly higher than that of the other extracts.

Conclusion

The experimental data revealed that Ranunculus pedatus showed significant wound healing and anti-inflammatory effect.     

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.     

Bioassay-guided isolation of anti-inflammatory, antinociceptive and wound healer glycosides from the flowers of Verbascum mucronatum Lam.

Ethnopharmacological relevance

The leaves, flowers and whole aerial parts of Verbascum L. species have been used to treat respiratory problems, haemorrhoids and other types of inflammatory conditions in traditional Turkish medicine.

Aim of the study

In order to evaluate this traditional information, the anti-inflammatory, antinociceptive and wound healing activities of Verbascum mucronatum Lam. which is used as haemostatic in Turkish folk medicine were investigated.

Materials and methods

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. Moreover, the wound healing potential of the plant were evaluated by using in vivo wound healing experimental models, i.e. incision and excision models on mice and rats, were comparatively assessed with a reference ointment Madecassol^®. Skin samples were also evaluated histopathologically.

Results

The results of these experimental studies exhibited that Verbascum mucronatum displays anti-inflammatory, antinociceptive and wound healing activities. Through bioassay-guided fractionation and isolation procedures four iridoid glucosides, ajugol (1), aucubin (2), lasianthoside I (3), catalpol (4), two saponins, ilwensisaponin A (5) and C (6) and a phenylethanoid glycoside, verbascoside (7) were isolated and their structures were elucidated by spectral techniques. Verbascoside (7) was found to possess significant wound healing activity as well as antinociceptive and anti-inflammatory potentials, per os without inducing any apparent acute toxicity or gastric damage.

Conclusion

The experimental study revealed that Verbascum mucronatum displays remarkable antinociceptive, anti-inflammatory and wound healing activities.     

Wound healing acceleration effect of endemic Ononis species growing in Turkey

Aim of the study

Ononis L. species have been used for healing of wounds, eczema and rheumatic complaints, against skin cancer and lesions and topically used as antiseptic and antimicrobial agent in folk medicine. In the present study, the aqueous and methanolic extracts of three endemic Ononis species growing in Turkey, including Ononis sessilifolia Bornm., Ononis basiadnata Hub.-Mor. and Ononis macrosperma Hub.-Mor. were assessed for their in vivo wound healing and anti-inflammatory activities.

Materials and methods

In vivo wound healing activities of the plants were evaluated by using linear incision and circular excision experimental models subsequently histopathological analysis. The healing potential was comparatively assessed with a reference ointment Madecassol^®, which contains 1% extract of Centella asiatica. 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 aqueous and ethanolic extracts of the aerial parts of O. macrosperma demonstrated the highest activity in both wound models and significant anti-inflammatory activity in acetic acid-induced increase in capillary permeability test. Furthermore, ethanolic extract of the aerial parts of O. sessilifolia showed anti-inflammatory effect but was devoid of wound healing activity. The rest of the species did not show remarkable wound healing effect. Ethyl acetate fraction of O. macrosperma ethanolic extract demonstrated significant but reduced wound healing activity, which revealed a possible combined effect. The results of histopathological examination supported the outcome of linear incision and circular excision wound models.

Conclusion

The experimental study revealed that the aerial parts of O. macrosperma display remarkable wound healing and anti-inflammatory activities.     

An ethnopharmacological study on Verbascum species: From conventional wound healing use to scientific verification

Aim of the study

The leaves, flowers, and whole aerial parts of Verbascum L. (Scrophulariaceae) species are used to treat eczema and other types of inflammatory skin conditions and as a desiccant for wounds in Turkish traditional medicine. In the present study, the methanolic extracts of 13 Verbascum species growing in Turkey, including Verbascum chionophyllum Hub.-Mor., Verbascum cilicicum Boiss., Verbascum dudleyanum (Hub.-Mor.) Hub.-Mor., Verbascum lasianthum Boiss., Verbascum latisepalum Hub.-Mor., Verbascum mucronatum Lam., Verbascum olympicum Boiss., Verbascum pterocalycinum var. mutense Hub.-Mor., Verbascum pycnostachyum Boiss. & Heldr., Verbascum salviifolium Boiss., Verbascum splendidum Boiss., Verbascum stachydifolium Boiss. & Heldr and Verbascum uschackense (Murb.) Hub.-Mor. were assessed for their in vivo wound healing activity.

Material and methods

In vivo wound healing activity of the plants were evaluated by linear incision and circular excision experimental models subsequently histopathological analysis. The healing potential was comparatively assessed with a reference ointment Madecassol^®, which contains 1% extract of Centella asiatica.

Results

The methanolic extracts of Verbascum olympicum, Verbascum stachydifolium and Verbascum uschackense demonstrated the highest activities on the both wound models. Moreover, the methanolic extracts of Verbascum latisepalum, Verbascum mucronatum, and Verbascum pterocalycinum var. mutense were found generally highly effective. On the other hand, the rest of the species did not show any remarkable wound healing effect. Results of the present study support the continued and expanded utilization of these plant species employed in Turkish folk medicine.

Conclusion

The experimental study revealed that Verbascum species display remarkable wound healing activity.     

Enhancement of wound healing by topical application of Scorzonera species: determination of the constituents by HPLC with new validated reverse phase method

Ethnopharmacological relevance Scorzonera species are mainly used against arteriosclerosis, kidney diseases, hypertension, diabetes mellitus and rheumatism, as well as for pain relief and wound healing in Turkish folk medicine. In the current study, aerial parts and roots of Scorzonera cinerea, Scorzonera incisa, Scorzonera latifolia, Scorzonera mollis ssp. szowitsii, Scorzonera parviflora, and Scorzonera tomentosa collected from different regions of Anatolia and yak? sak?z? which is prepared by drying the latex is obtained from the roots of Scorzonera latifolia were evaluated for their wound healing and anti-inflammatory effects.

Materials and methods

In vivo wound healing activity of the plants was evaluated by linear incision and circular excision wound models. Hydroxyproline content of the treated tissues was also assessed. Acetic acid-induced capillary permeability test was used for anti-inflammatory activity. For the determination of the active constituents, a new high performance liquid chromatographic method was developed. Precision of the developed 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 Scorzonera latifolia, Scorzonera mollis ssp. szowitsii demonstrated the highest activities. The main compound was detected as chlorogenic acid both in roots and aerial parts of the most Scorzonera species and hyperoside or rutin was determined as one of the main component of the aerial parts.

Conclusion

The experimental study revealed that methanol-water extracts from aerial parts of Scorzonera latifolia and Scorzonera mollis ssp. szowitsii were found to have better activity on the wound healing compared to the other extracts and control groups. This might be due to the combined effect of the constituents present in the extracts.     

Efficacy of Daphne oleoides subsp. kurdica used for wound healing: identification of active compounds through bioassay guided isolation technique

Ethnopharmacological relevance

In Turkish traditional medicine, the aerial parts of Daphne oleoides Schreber subsp. kurdica (DOK) have been used to treat malaria, rheumatism and for wound healing.The aim was to evaluate the ethnopharmacological usage of the plant using in vivo and in vitro pharmacological experimental models, and to perform bioassay-guided fractionation of the 85% methanolic extract of DOK for the isolation and identification of active wound-healing component(s) and to elucidate possible mechanism of the wound-healing activity.

Materials and methods

In vivo wound-healing activity was evaluated by the linear incision and the circular excision wound models. Anti-inflammatory and antioxidant activities, which are known to support the wound healing process, were also assessed by the Whittle method and the DPPH (2,2-diphenyl-1-picrylhydrazyl) radical-scavenging assays, respectively. The total phenolic content of the extract and subextracts was estimated to establish any correlation between the phenolic content and the antioxidant activity. The methanolic extract of DOK was subjected to various chromatographic separation techniques leading to the isolation and identification of the active component(s). Furthermore, in vitro hyaluronidase, collagenase and elastase enzymes inhibitory activity assays were conducted on the active components to explore the activity pathways of the remedy.

Results

After confirmation of the wound-healing activity, the methanolic extract was subjected to successive solvent partitioning using solvents of increasing polarity creating five subextracts. Each subextract was tested on the same biological activity model and the ethyl acetate (EtOAc) subextract had the highest activity. The EtOAc subextract was subjected to further chromatographic separation for the isolation of components 1, 2 and 3. The structures of these compounds were elucidated as daphnetin (1), demethyldaphnoretin 7-O-glucoside (2) and luteolin-7-O-glucoside (3). Further in vivo testing revealed that luteolin-7-O-glucoside was responsible for the wound-healing activity of the aerial parts. It was also found to exert significant anti-inflammatory, antioxidant, anti-hyaluronidase and anti-collagenase activities.

Conclusion

The present study explored the wound-healing potential of Daphne oleoides subsp. kurdica. Through bioassay-guided fractionation and isolation techniques, luteolin-7-O-glucoside was determined as the main active component of the aerial parts. This compound exerts its activity through inhibition of hyaluronidase and collagenase enzymes activity as well as interfering with the inflammatory stage.     

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.     

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.     

The healing and anti-scar effects of astragaloside IV on the wound repair in vitro and in vivo

Ethnopharmacological relevance

Astragaloside IV is the chief ingredient of Radix Astragali, which has been used in the Traditional Chinese Medicine as a major component of many polyherbal formulations for the repair and regeneration of injured organ and tissues. This study is to investigate the influence of astragaloside IV on both of the wound healing and scar formation.

Materials and methods

For the in vitro evaluation, the influence of the astragaloside IV in the wound scratch test of keratinocytes and the secretion of transforming growth factor-β1, a key factor contributing to scar formation were determined. With the rat skin excision model, the in vivo regulation of astragaloside IV on wound closure, angiogenesis and collagen disposition were also evaluated.

Results

Astragaloside IV was shown to significantly promote the migration of keratinocytes in wound scratching assay. The superior effect of Astragaloside IV was observed at 100 μmol/L, in which the recover rates was increased with 2 and 3 folds after 48 h and 96 h respectively than that of blank control (P < 0.01). Animal skin closure measurement showed that astragaloside IV could stimulate the wound healing, e.g. with 21% recover in contrast to the 8% of blank control at the 6th day. Biomechanic and Masson's trichrome stain analysis indicated that astragaloside IV may improve the strength of the repaired skin and promoted the angiogenesis and collagen synthesis. Meanwhile, the picrosirius-sirus red stain and Elisa test definitely showed the anti-scar effects of astragaloside IV by decreasing the levels of collagen I/III and TGF-β1 secretion by firbroblasts with a dose-dependent manner (25-100 μmol/L).

Conclusions

Astragaloside IV was shown a promising natural product with both healing and anti-scar effects for wound treatment. These results give the evidence for the application of astragaloside IV in the treatment of injury.     

The essential oil of Croton zehntneri and trans-anethole improves cutaneous wound healing

Ethnopharmacology relevance

Croton zehntneri is a Euphorbiaceae species native to northeastern Brazil, where teas and beverages made from Croton zehntneri leaves are used as healing agents. To our knowledge, there is no experimental study supporting this claim of pharmacological activity.

Materials and methods

Full-thickness excisional wounds were made in the left and right sides of the dorsum of anesthetised Swiss mice, and a topical pharmaceutical formulation, developed by including essential oil extracted from the leaves of Croton zehntneri (2% and 20% EOCz) in pluronic-127 (PF-127), was administered to mice twice daily for 15 days post-wounding. To evaluate the contribution of trans-anethole, the major constituent of EOCz (85.7%), in the wound healing activity of EOCz, the effect of the topical administration of trans-AT on wound tissue repair was also evaluated and compared to other groups. A macroscopic analysis of swelling and exudates was performed and scored as 0 (missing), 1 (light), 2 (moderate) and 3 (intense). The number of capillaries and leukocytes was counted in hematoxylin and eosin (HE)-stained sections of the injured tissue. For extracellular matrix remodelling analysis, fibroblasts and collagen fibres present in the photomicrography of the Masson’s Trichrome (MT)-stained sections were counted. Each experimental group comprised six mice.

Results

At day 3 post-wounding, it was observed that treatment with 20% EOCz greatly reduced the swelling and exudates with a similar magnitude to the dexamethasone treatment. The inflammatory cell infiltration and angiogenesis were not altered by either the EOCz- or trans-AT treatments. In contrast, an acceleration of the wound closure was observed, with an enhanced number of fibroblasts and collagen fibres in both the 20% EOCz- and trans-AT-treated mice.

Conclusion

Our data indicate that EOCz exerts significant wound healing activity, demonstrating its relevant therapeutic potential.     

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.     

Antinociceptive, anti-inflammatory and wound healing features in animal models treated with a semisolid herbal medicine based on Aleurites moluccana L. Willd. Euforbiaceae standardized leaf extract: Semisolid Herbal

Ethnopharmacological relevance

Aleurites moluccana L. (Willd) Euforbiaceae is a native tree of Indonesia and India that has become acclimatized and well-adapted to the South and Southwest of Brazil. It is commonly used in traditional medicine to treat pain, fever, inflammation, asthma, hepatitis, headache, gastric ulcer, cuts, skin sores and other ailments. The oral antinociceptive effects of standardized 70:30 (v/v) ethanol:water spray dried extract of A. moluccana leaf, as well as its flavonoids 2?-O-rhamnosylswertisin (I) and swertisin (II), have previously been reported.

Aim

The aim of this study was to develop a stable and effective semisolid herbal medicine for topical use in the treatment of pain, inflammation and wound healing, containing 0.5 and 1.0% of standardized dried extract of A. moluccana.

Materials and methods

The chemical markers I and II were assayed by HPLC-UV analysis after extraction by matrix solid dispersion phase (MSDP) followed analytical validation as ICH Guidelines. The semisolid preparations of Hostacerin CG^® vehicle containing 0.5 and 1.0% of dried extract of A. moluccana were submitted to stability studies (180 day of accelerated and long-term studies). The phytomedicine semisolid was analysed in croton oil-induced ear oedema model in mice, in the healing process, using the excisional wound model in rats, and to prevent mechanical sensitization following plantar incision in rats in the postoperative model of pain.

Results

The MSDP method showed average recovery of 101.6 and 105.7% for I and II, respectively, with good precision (RSD<2.0%) and selectivity, without interference of the excipients. The formulations were approved in the stability studies, maintaining conformity after 180 day of accelerated and long-term studies, with variation<10% in the analytical parameters. The phytomedicine reduced the ear oedema in 37.6±5.7% and 64.8±6.2%, for 0.5 and 1.0% of dried extract, respectively. The formulation also accelerated the healing process by up to 50.8±4.1% and 46.0±4.0% at 0.5 and 1.0% of extract, respectively, and both amounts were capable of preventing the development of mechanical sensitization following plantar incision in rats.

Conclusions

The MSDP followed by HPLC-UV analytical method was appropriate for the quality control of the topical phytomedicine based on A. moluccana. The formulation developed at 0.5 and 1.0% of A. moluccana dried extract proved to be effective as an analgesic, anti-inflammatory and wound healing in the pre-clinical studies, which is in agreement with the ethnopharmacological data.     

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.     

Evaluation of in-vivo wound healing activity of Hypericum patulum (Family: hypericaceae) leaf extract on different wound model in rats

The methanol extract of Hypericum patulum Thumb. leaves were investigated for the evaluation of their wound healing potential on different experimental models of wounds in rats. The methanol extract of leaves (HPM), in the form of an ointment with two different concentrations (5% and 10% w/w ointment of leaf extract in simple ointment base) was evaluated for wound healing potential in an excision wound model and an incision wound model in rats. Both concentrations of the methanol extract ointment showed significant responses in both the wound types tested when compared with the control group. The effect produced by the extract ointment, in terms of wound contracting ability, wound closure time, regeneration of tissues at wound site, tensile strength of the wound and histopathological characteristics were comparable to those of a standard drug nitrofurazone ointment.     

In vitro growth stimulatory and in vivo wound healing studies on cycloartane-type saponins of Astragalus genus

Aim of the study

The present study was undertaken to evaluate the wound healing effects of the four chief saponins of Astragalus species [cycloastragenol (CA), astragaloside IV (AG), cyclocephaloside I (CCI) and cyclocanthoside E (CCE)].

Material and methods

Effects of cell viability and proliferation of the isolated compounds were evaluated by the MTT assay on human keratinocyte. The wound healing activity was studied by using in vitro wound healing, proliferation and migration scratch assay. In order to see in vivo effectiveness of the compounds, an animal study with Sprague-Dawley male rats at the age of 12 weeks was carried out, and then the main histological outcomes were investigated to observe reepithelization, neovascularization, and presence of inflammatory cells, granulation tissue amount and maturation.

Results

All the compounds increased both fibroblast proliferation and migration, but the effects were much superior for CA at 1 ng/ml concentration. Among the compounds, based on the histological findings, 5% CA preparation was found to be the most remarkable in vivo wound healing agent showing greater cell density, more regularly organized dermis and more newly formed blood vessels.

Conclusion

Results of this study indicate that the cycloartane-type saponins are the principal constituents responsible for wound healing activities of the roots of Astragalus species substantiating its use in traditional medicine.