Jelena Živković

Ethnobotanical study on traditional use of medicinal plants in South-Western Serbia, Zlatibor district.

ETHNOPHARMACOLOGICAL RELEVANCE This paper provides significant ethnobotanical information on medicinal plant uses in the Zlatibor district, South-Western Serbia. MATERIALS AND METHODS A survey was performed using questionnaires with 220 informants (mean age 47, 79% female, 21% male). In addition, the use value and the relative importance of species were determined and the informant consensus factor was calculated for the medicinal plants included in the study. Intended plants usage was compared with previous ethnobotanical literature, with reference to the neighboring areas of Zlatibor district. RESULTS The informants provided data for 69 medicinal plants belonging to 36 families. Rosaceae, Lamiaceae and Asteraceae were the predominant locally used families. The species with the highest use value were Mentha piperita, Matricaria chamomilla, Hypericum perforatum and Achillea millefolium. The most frequently reported medicinal uses were ones for treating gastrointestinal ailments, respiratory problems and skin diseases. Usually, the administration was primarily oral followed by topical applications. All different plant parts were utilized, however leaves were the most exploited parts of the plants. CONCLUSIONS Folk medicine in South-Western Serbia, Zlatibor district is intended mainly as a mode of primary health care in healing of minor illnesses. The results indicate a slight reduction in the ethnobotanical and medical knowledge in this area, when compared with neighboring regions.

Addition of blood to a phycogenic bone substitute leads to increased in vivo vascularization

The present study aimed to analyze the effects of the addition of blood to the phycogenic bone substitute Algipore(®) on the severity of in vivo tissue reaction. Initially, Fourier-transform infrared spectroscopy (FTIR) of the bone substitute was conducted to analyze its chemical composition. The subcutaneous implantation model in Balb/c mice was then applied for up to 30 d to analyze the tissue reactions on the basis of specialized histochemical, immunohistochemical, and histomorphometrical methods. The data of the FTIR analysis showed that the phycogenic bone substitute material is mainly composed of hydroxyapatite with some carbonate content. The in vivo analyses revealed that the addition of blood to Algipore(®) had a major impact on both angiogenesis and vessel maturation. The higher vascularization seemed to be based on significantly higher numbers of multinucleated TRAP-positive cells. However, mostly macrophages and a relatively low number of multinucleated giant cells were involved in the tissue reaction to Algipore(®). The presented data show that the addition of blood to a bone substitute impacts the tissue reaction to it. In particular, the immune response and the vascularization were influenced, and these are believed to have a major impact on the regenerative potential of the process of bone tissue regeneration.

Characterisation of volatiles in dried white varieties figs (Ficus Carica L.)

The aromatic profile of volatiles in dried figs varieties Bružetka Bijela and Zimnica were characterised by headspace solid-phase (HS-SPME) procedure with gas chromatography–mass spectrometry analysis (GC-MS). The volatile compounds were distributed by distinct chemical classes, including alcohols, aldehydes, esters, terpenic compounds, and other compounds. The figs were dried in a pilot plant cabinet dryer. Prior to drying process, figs were pre-treated by sulphur dioxide, immersed in solution of citric acid and ascorbic acid, respectively. Several mathematical thin-layer drying models, available in the literature, were fitted to experimental data of figs, implementing non-linear regression analysis techniques. The results showed that pre-treatments of figs decrease significantly the drying time. The best thin-layer drying model in terms of fitting performance was Wang and Singh model. The major volatile compound in dried figs was benzaldehyde. After benzaldehyde, the most abundant aldehyde in dried figs was hexanal. The comparison among dried figs showed the highest abundance of aldehydes, in general, in non-treated (control) dried figs compared to pre-treated samples. Furthermore, ascorbic acid was the most efficient in preserving esters and alcohols in case of Bružetka Bijela, whereas in case of Zimnica, sulphur dioxide was in advance compared to ascorbic acid. Ethyl acetate was the most abundant ester found in dried figs. Among other compounds, 2-butanone,3-hydroxy was the most abundant identified volatiles. Linalool, as the only identified terpen, was in case of both dried fig varieties, preserved by immersion into ascorbic acid. The immersion into citric acid has not been so successful in volatiles conservation.

Phenolic profiling of Veronica spp. grown in mountain, urban and sandy soil environments

Veronica (Plantaginaceae) genus is widely distributed in different habitats. The phenolic compounds of Veronica montana, Veronica polita and Veronica spuria were tentatively identified by HPLC-DAD-ESI/MS. The phenolic profiles showed that flavones were the major compounds (V. montana: 7 phenolic acids, 5 flavones, 4 phenylethanoids and 1 isoflavone; V. polita: 10 flavones, 5 phenolic acids, 2 phenylethanoids, 1 flavonol and 1 isoflavone; V. spuria: 10 phenolic acids, 5 flavones, 2 flavonols, 2 phenylethanoids and 1 isoflavone). V. spuria possessed the highest contents in all groups of phenolic compounds, except flavones, which did not show differences among the assayed species. Overall, these species might be considered good sources of phenolic compounds for industrial or pharmacological applications.

In vivo and in vitro antioxidant effects of three Veronica species

The aim of the present study was to examine the antioxidant activity of three Veronica species (Plantaginaceae). The antioxidant potential of various extracts obtained from aerial flowering parts was evaluated by DPPH-free (1,1-diphenyl-2-picrylhydrazyl-free) radical scavenging activity and ferric-reducing antioxidant power assays. Considerable antioxidant activity was observed in the plant samples (FRAP values ranged from 0.97 to 4.85 mmol Fe2+/g, and DPPH IC50 values from 12.58 to 66.34 µg/ml); however, these levels were lower than the activity of the control compound butylated hydroxytoluene (BHT) (FRAP: 10.58 mmol Fe2+/g; DPPH IC50: 9.57 µg/ml). Also, the in vivo antioxidant effects were evaluated in several hepatic antioxidant systems in rats (activities of glutathione peroxidase, glutathione reductase, peroxidase, catalase, xanthine oxidase, glutathione content and level of thiobarbituric acid reactive substances) after treatment with different Veronica extracts, or in combination with carbon tetrachloride (CCl4). Pretreatment with 100 mg/kg b.w. of Veronica extracts inhibited CCl4-induced liver injury by decreasing TBA-RS level, increasing GSH content, and bringing the activities of CAT and Px to control levels. The present study suggests that the extracts analyzed could protect the liver cells from CCl4-induced liver damage by their antioxidative effect on hepatocytes.

Apatite formation on nanomaterial calcium phosphate/poly-dL-lactide-co-glycolide in simulated body fluid

Simulated body fluid (SBF) is an artificial fluid which has ionic composition and ionic concentration similar to human blood plasma. Purpose This paper compares the interaction between the nanomaterial containing calcium phosphate/poly-DL-lactide-co-glycolide (N-CP/PLGA) and SBF, in order to investigate whether and to what extent inorganic ionic composition of human blood plasma leads to the aforementioned changes in the material. Methods N-CP/PLGA was incubated for 1, 2, 3, and 5 weeks in SBF. The surface of the material was analyzed on SEM-EDS and FTIR spectrometer, while SBF was subjected to pH and electrical conductivity measurement. Results Our results indicate that dissolution of the polymer component of the material N-CP/PLGA and precipitation of the material similar to hydroxyapatite on its surface are based on the morphologic changes seen in this material. Conclusions The mechanism of the apatite formation on the bioceramic surface was intensively studied and was considered crucial in designing the new biomaterials. The results obtained in this work indicate that N-CP/PLGA may be a good candidate for application to bone regeneration.

Fatty Acid Profiles of Four Wild Mushrooms and Their Potential Benefits for Hypertension Treatment

Mushrooms are useful in balancing human diet and treating different health problems. The main determinant of the fluidity of erythrocyte membranes and rheologic properties of blood is the lipid composition of the membrane, which directly depends on lipids present in the diet. Lipid profiles of 4 European wild edible mushrooms, as well as the ability of mushroom lipids to modify the fluidity of erythrocyte membrane, were examined by using gas chromatography-mass spectometry, gas chromatography-flame ionization detector, and electron paramagnetic resonance spin probing technique. Lipids from 2 species--Macrolepiota procera and Collybia platyphylla--provoked an increase in erythrocyte membrane fluidity. Therefore, introduction of these and other wild mushrooms with similar lipid profiles to the human diet could be very beneficial in the treatment of hypertension and other cardiovascular diseases related to decreased fluidity of erythrocyte membranes.

Effects of bone tissue engineering triad components on vascularization process: comparative gene expression and histological evaluation in an ectopic bone-forming model

ABSTRACT Vascular development has a great significance in the osteogenic process and in bone tissue engineering (BTE). BTE is based on various combinations of three principal types of components: biomaterials as scaffolds, regulatory signals and cells. The aim of our study was to evaluate, at gene expression and histological level, the effect of BTE triad components on the vascularization process in an ectopic bone-forming model in mice. Bone mineral matrix (BMM) was used as a scaffold and a carrier, platelet-rich plasma (PRP) as a source of regulatory signals and adipose stem cells (ASCs) as a source of cells for endothelial differentiation, in order to show how and to what extent the biological enrichment of BMM influences the outcome of the osteogenic process and its key precondition, vascularization. Implants composed of BMM, PRP and ASCs in vitro induced into endothelial cells (EPB implants) and implants composed of BMM and PRP (PB implants) were compared with implants composed of BMM only (B implants). More pronounced endothelial-related gene expression and stronger VCAM-1 (vascular cell adhesion molecule-1) immunoexpression were observed in EPB implants in comparison with PB and B ones at later time points of the in vivo experimental period. Osteopontin gene expression and immunoexpression of osteopontin were significantly higher in EPB compared to PB and B implants. Therefore, addition of ASCs combined with PRP to BMM improved the vascularization process in the ectopic bone-forming model, which makes this BTE composition the most favourable among the examined types of implants for application in BTE.

Evaluation of angiogenic and neuroprotective potential of different extracts from three Veronica species

Methanol and 70% acetone extracts from Veronica jacquinii, Veronica urticifolia and Veronica teucrium were evaluated regarding their neuroprotective/antioxidant effects on the human neuroblastoma SH-SY5Y cell line. All extracts exhibited modest protective activity by increasing cell survival in cells stressed with sodium nitroprusside (9–12%) and hydrogen peroxide (16–21%) compared to non-treated cells. These activities were accompanied by reductions in the amount of superoxide radicals and index of lipid peroxidation. Extracts were further analysed for the content of total phenolics, phenylpropanoid glycosides and iridoids, and the major compounds acteoside and aucubin. The highest amount of total phenolics was observed in V. jacquinii, while V. teucrium was richest in total iridoids, acteoside and aucubin. Angiogenic properties on EA.hy926 human endothelial cells were also examined. At the highest non-toxic concentration (25 µg/ml), the tested extracts inhibited spontaneous tube formation of endothelial cells on the extracellular matrix, implying their possible antiangiogenic activity. The most potent inhibitory effects were shown by methanol extract of V. jacquinii and aqueous acetone extract of V. teucrium. The extracts did not significantly change the adhesive or migratory capacity of EA.hy926 cells. Considering the traditional use of Veronica species, these results suggest a need for further assessment of their supposed wound-healing properties.

The Influence of Nanomaterial Calcium Phosphate/poly-(dl-lactide-co-glycolide) on Proliferation and Adherence of HeLa Cells

Nanomaterials are nowadays widely used in various fields of biomedicine. Before the application of biomaterials they have to be tested and fulfill some criteria. The first tests that should be performed after synthesis of some new nanomaterial with potential application in biomedical fields are biocompatibility tests. The aim of this study was to examine the influence of nanomaterial calcium phosphate/poly-(dl-lactide-co-glycolide) (CP/PLGA) on proliferation and adherence of HeLa cells in culture. For both proliferation and adherence examination, cells were treated with suspension of nanomaterial CP/PLGA at concentrations 5, 50 and 500 µg/mL, made in cell culture medium. There were four different types of treatment: (1) cells incubated with CP/PLGA for 3 days; (2) cells incubated with CP/PLGA for 6 days; (3) cells incubated first with CP/PLGA for 3 days and then for the next 3 days with medium and (4) cells incubated first with medium for 3 days and then for the next 3 days with CP/PLGA. Three days after incubation of HeLa cells with different concentrations of CP/PLGA nanoparticles’ suspension, the concentration of 5 µg/mL had mild inhibitory effect on proliferation. Increasing CP/PLGA concentration, there was stimulatory effect on cells’ proliferation. With prolonged incubation period, this dose dependence is lost. The highest adherence of HeLa cells was observed when cells were incubated with the highest examined concentration of CP/PLGA suspension, in both 3-day and 6-day incubation period. Based on the results obtained in our study, we can conclude that the effect of the suspension of nanomaterial CP/PLGA on proliferation and adherence of HeLa cells depends on the duration of incubation with the cells as well as the material concentration and type of the treatment.