Hikmet Keles

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.

Beneficial effects of Aesculus hippocastanum L. seed extract on the body's own antioxidant defense system on subacute administration.

AIM OF THE STUDY Seeds of Aesculus hippocastanum L. have long been used in European phytotherapy to treat inflammatory and vascular problems. In Turkish folk medicine, tea prepared from the crushed seeds was used to pass kidney stone and against stomach ache, while a fraction of seed was swallowed to alleviate hemorrhoids symptoms. In order to evaluate the in vivo effects of escin mixture from Aesculus hippocastanum seed on the blood and tissue antioxidant defense systems in standard pellet diet (SPD) and in high-fat diet (HFD) consumed male mice. MATERIALS AND METHODS Escin mixture was obtained from the ethanol extract of seeds. Escin mixture was administered orally to male mice fed either standard pellet diet (SPD) or high-fat diet (HFD) at 100mg/kg doses daily for 5 weeks and the tissue (liver, kidney and heart) and blood samples were collected at the end of experimental period. The effect of escin mixture on the plasma antioxidant activity; blood and tissue malondialdehyde (MDA) and reduced glutathione (GSH) levels; erythrocyte and tissue superoxide dismutase (SOD) and catalase activity (CAT) in SPD and HFD consumed animals were experimentally studied. RESULTS Escin mixture prohibited the adverse effects of oxidative stress and showed a protective effect on the liver architecture both in SPD and HFD consumed male mice. Escin mixture prohibited the adverse effects of oxidative stress and showed a protective effect on the liver architecture both in SPD and HFD consumed male mice. Combined administration of high-fat diet with escin mixture decreased blood (p<0.01), liver (p<0.01), kidney (p<0.05), and heart (p<0.05) of MDA, liver SOD (p<0.01) and CAT (p<0.05) levels and increased blood (p<0.01) and liver GSH (p<0.001) levels in mice. CONCLUSION The present results indicate that Aesculus hippocastanum increase the antioxidative defense system of the body and prevent HFD-induced lipid peroxidation in male mice.

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.

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 Activity of Rubus sanctus Schreber (Rosaceae): Preclinical Study in Animal Models

Young shoots of Rubus species have been used for healing of wounds, infected insect bites and pimples in folk medicine for ages. In order to evaluate the wound healing activity of Rubus sanctus, four different extracts were prepared from the whole aerial parts of the plant by using n-hexane, chloroform, ethyl acetate and methanol, respectively. Incision wound healing model by using tensiometer on rats and excision model on mice were employed to assess the activity. Remarkable wound healing activity was observed with the ointment formulation of the methanol extract at 1% concentration on the mentioned models. The results of histopathological examination also supported the outcome of both incision and excision wound models. The wound healing effect was comparatively evaluated with a reference ointment Madecassol. The experimental data confirmed the ethnobotanical usage of R. sanctus.

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.

Appraisal on the wound healing and anti-inflammatory activities of the essential oils obtained from the cones and needles of Pinus species by in vivo and in vitro experimental models

ETHNOPHARMACOLOGICAL RELEVANCE According to ethnobotanical data, Pinus species have been used against rheumatic pain and for wound healing in Turkish folk medicine. Essential oils from the cones and needles of five different Pinus species (Pinus brutia Ten., Pinus halepensis Mill., Pinus nigra Arn., Pinus pinea L. and Pinus sylvestris L.) were evaluated for their in vivo wound healing and anti-inflammatory activities. MATERIALS AND METHODS In vivo wound healing activity of the ointments prepared from essential oils was evaluated by linear incision and circular excision experimental wound models subsequently histopathological analysis and hydroxyproline content. Furthermore, the essential oils were screened for anti-hyaluronidase activity. Additionally anti-inflammatory activity was assessed by using the method of Whittle, which is based on the inhibition of acetic acid-induced increase in capillary permeability. RESULTS The essential oils obtained from the cones of Pinus pinea and Pinus halepensis demonstrated the highest effects on the wound healing activity models. On the other hand, the rest of the essential oils did not show any significant wound healing and anti-inflammatory activities. CONCLUSION The experimental study revealed that essential oils obtained from the cones of Pinus pinea and Pinus halepensis display remarkable wound healing activity.

Increased DNA damage and oxidative stress in chickens with natural Marek's disease.

Oxidative stress contributes to the accumulation of genomic abnormalities, prevents cellular apoptosis, and also mediates immunosuppression resulting in tumor formation. Mareks Disease provides excellent opportunities for the study of herpesvirus-induced tumors both in experimental- and natural conditions. The aim of this study was to examine the effects of Mareks Disease (MD) on basal levels of DNA strand breaks and on the oxidative-antioxidative status of chickens with MD. White-Lohmann hens-fifteen infected with Mareks Disease Virus (MDV) and fifteen healthy-of same age and sex were included in this study. MD infection was diagnosed via clinical signs, gross- and micro-pathological findings and also by detection of viral antigens in feather follicle epithelium by the indirect immunoperoxidase method. Compared with healthy controls, DNA damage was greater and levels of malondialdehyde (MDA) and plasma protein carbonyl (PCO), and plasma concentration of nitric oxide metabolites (NOx) higher in the MD group. Furthermore, total antioxidant activities (AOAs) were found lowered and glutathione (GSH) levels reduced in the MD group compared to the control group. Significantly positive correlation was found between DNA damage, MDA, PCO, and NOx in the MD group. DNA strand breaks were found negatively associated with AOA and GSH concentrations in the MD group. Our results demonstrated that oxidative stress markers and DNA damage substantially increased in chickens with MD, which indicated that increased DNA damage levels might be related to the increased oxidative stress and reduced antioxidant 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 yakı sakızı 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.

Cholesterol-reducer, antioxidant and liver protective effects of Thymbra spicata L. var. spicata.

ETHNOPHARMACOLOGICAL RELEVANCE Thymbra spicata is a member of the Lamiaceae family; leaves of this plant have recently gained much popularity as a remedy to combat hypercholesterolaemia. AIM OF THE STUDY To evaluate the antihypercholesterolaemic, antioxidant and anti-steatohepatitic activities of the diethyl ether (DEE), ethyl acetate (EtOAc) and remaining aqueous (RA) extracts from Thymbra spicata var. spicata in mice. MATERIALS AND METHODS In this study, diethyl ether, ethyl acetate and remaining aqueous extracts of Thymbra spicata L. var. spicata P.H.Davis (Lamiaceae) were evaluated for the effects on the plasma total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), triglyceride (TG) and glucose; blood malondialdehyde (MDA) and reduced glutathione (GSH); erythrocyte superoxide dismutase (SOD) and catalase activity (CAT) in mice fed with high-fat diet (HFD). RESULTS The HFD induced an increase in plasma TC, TG, LDL, MDA concentrations compared to control group. However, administration of DEE with HFD reduced TC, LDL, TG and MDA concentrations, while increased HDL concentration, as well as GSH, SOD and CAT activities compared to HFD. The other extract from the plant was RA, which also showed a similar activity profile with DEE except CAT. On the other hand, administration of EtOAc extract with HFD decreased plasma TC, TG and MDA, while GSH concentration was increased. Histopathologically, best liver conditions were observed in DEE and lesser in RA extracts. Based on the results obtained in this investigation it is suggested that the DEE and partially RA extracts of Thymbra spicata var. spicata displayed significant antihypercholesterolaemic, antioxidant and anti-steatohepatitic activities. HPLC analysis of the DEE extract of Thymbra spicata var. spicata revealed the presence of carvacrol (44.13%). CONCLUSION The observed cholesterol-reducer, antioxidant and liver protective effects of the DEE and partially of RA extracts of Thymbra spicata which contain mainly carvacrol indicates that these extracts possess some potential medicinal value and explain their ethnomedical use.