Chemical composition determination and evaluation of the antibacterial activity of essential oils from Ruellia asperula (Mart. Ex Ness) Lindau and Ruellia paniculata L. against oral streptococci.

This study investigated the chemical composition and evaluated the antibacterial and antibiofilm activities of essential oils (EOs) extracted from Ruellia asperula (EORA) and Ruellia paniculata (EORP) against oral streptococci. The EO constituents were analyzed by gas chromatography/mass spectrometry. The antimicrobial potential of EOs was evaluated using the minimum inhibitory concentration, minimum bactericidal concentration, and time-kill determination. Furthermore, the quantification of total biomass and the number of viable cells in the biofilms were evaluated. The major constituents of EORA were cariophylla-4(12)-8-(13)-dien-5ß-ol (14.1%), (ß)-caryophyllene (22.7%), and caryophyllene oxide (29.4%). For EORP, the major constituents were (ß)-caryophyllene (11.0%), spathulenol (13.1%), and δ-amorphene (14.9%). The tested EOs exhibited antibacterial activity against planktonic growth and biofilm formation. Thus, the EOs from R. asperula and R. paniculata prove to be promising alternatives for bacterial growth control and biofilm formation prevention of oral streptococci.

Antifungal and Antioxidant Activities of Vernonia Chalybaea Mart. ex DC. Essential Oil and their Major Constituent β -caryophyllene

Abstract: This article describes the chemical composition of Vernonia chalybaea essential oil, and investigates its antimicrobial, antioxidant and hemolytic activities. The evaluation of the antifungal activity was performed by the broth microdilution method using strains of yeasts and dermatophytic fungi. The checkerboard technique to find antimicrobial modulatory effects was performed using ketoconazole as standard drug. The antioxidant activity was evaluated by DPPH scavenging assay and β-carotene/linoleic-acid system. The toxicity was characterized by the brine shrimp lethality test and hemolysis bioassays. The essential oil was obtained by hydrodistillation and analyzed by GC-MS method, showing to be rich in the sesquiterpenes β-caryophyllene (39.06%) and bicyclogermacrene (19.69%), and also demonstrated a relevant antifungal activity against strains of Trichophyton rubrum. In the modulatory activity assay, the essential oil of V. chalybaea and β-caryophyllene demonstrated a synergistic interaction with ketoconazole, with increasing of its antifungal action. The antioxidant activity was evidenced mainly by β-carotene/linoleic acid system, with IC50 value of 35.87 ± 0.32 µg/mL. The results suggest that V. chalybaea essential oil and β-caryophyllene are valuable natural medicinal agents with antioxidant and antimicrobial activities.

Tocoyena sellowiana extract decreases bone loss in an experimental model of periodontitis in rats: Putative role for cyclooxygenase-2 and IL-1? inhibition.

Tocoyena sellowiana (Cham. & Schltdl.) K.Schum is one of the most important families of Brazilian medicinal plants. This study aimed to evaluate the effect of Tocoyena sellowiana (Cham. & Schltdl.) K.Schum ethanolic extract in a pre-clinical trial of periodontitis and to investigate possible mechanisms underlying such effects. Periodontitis was induced in Wistar rats by placing a nylon thread ligature around second upper left molars for 11 days. Rats received (per os) Tocoyena sellowiana (0.1, 1 or 10?mg?kg) or vehicle 1?h before ligature and daily until day 11. Macroscopic, histopathological, and COX-2 immunohistochemical analyses were performed to evaluate the periodontium. The gingival tissue was used to quantify the myeloperoxidase (MPO) activity and interleukin (IL)-1? levels by ELISA. Blood samples were collected to evaluate bone-specific alkaline phosphatase (BALP), the dosage of creatinine, aspartate and alanine transaminases. The liver, kidneys, spleen, and body mass variations were also evaluated. Tocoyena sellowiana decreased bone loss, reduced MPO, IL-1? levels as well as COX-2 immunostaining, and increased BALP activity. Moreover, Tocoyena sellowiana did not alter organs nor body weight. Tocoyena sellowiana reduced bone loss in rats and its efficacy was at least partially dependent upon both IL-1? and cyclooxygenase-2 inhibition.

Effect of the triterpene 3ß, 6ß, 16ß-trihydroxylup-20(29)-ene isolated from the leaves of Combretum leprosum Mart. on cutaneous wounds in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Combretum leprosum Mart. is a native species of the Caatinga, an important biome in the Brazilian semi-arid region. This species is commonly used in Brazil as a healing agent, as well as for the treatment of skin diseases. AIM OF THE STUDY: This study investigated the healing potential of the ethanolic extract (EECL) and the bioactive triterpene 3ß, 6ß, 16ß-trihydroxylup-20 (29)-ene (CLF-1) isolated from the leaves of C. leprosum. MATERIALS AND METHODS: Skin wounds (1cm(2)) were created in the dorsal zone of mice with a scalpel blade number 15. The treatment consisted in a daily topical application of 100 µl of 150 mM NaCl, EECL and CLF-1 (at 10 µg/100 µl) for 12 days. The lesions were then macro and microscopically evaluated. RESULTS: On postoperative day (POD) 2, the lesions treated with EECL and CLF-1 showed a moderate presence of vessels of the granulation tissue progressing in the dermis. The same effect was not observed in the control group. The treatment with EECL and CLF-1 stimulated angiogenesis, resulting in a rapid deposition of extracellular matrix (ECM). Moreover, the animals treated with EECL and CLF-1 showed smaller lesions on POD 7, primarily due to the contraction in the reticular dermis induced by organization of myofibroblasts, which was not observed in the group treated with NaCl. In addition, the lesions treated with EECL and CLF-1 showed ECM restructuration and presence of epithelium coating, which was not observed in the group treated with NaCl, in which the lesions showed no epithelial lining, suggesting delayed healing. CONCLUSION: CLF-1 isolated from the leaves of C. leprosum may be considered to be an important molecule for the treatment of skin lesions. However, further investigations are necessary to establish its role in chronic lesions and to elucidate the mechanism of action involved in the cutaneous healing process. To the best of our knowledge, this is the first report describing the pro-healing activity of the ethanolic extract and the triterpene 3ß, 6ß, 16ß-trihydroxylup-20(29)-ene isolated from leaves of C. leprosum.

Antibacterial and antioxidant activities of derriobtusone A isolated from Lonchocarpus obtusus.

This study evaluated the effect of derriobtusone A, a flavonoid isolated from Lonchocarpus obtusus, on two important pathogenic bacteria, Staphylococcus aureus and Escherichia coli, as well as its antioxidant activity and toxicity. Planktonic growth assays were performed, and the inhibition of biofilm formation was evaluated. In addition, antioxidant activity was assessed by DPPH radical scavenging assay, ferrous ion chelating assay, ferric-reducing antioxidant power assay, and ß -carotene bleaching assay. Toxicity was evaluated by the brine shrimp lethality test. Results showed that derriobtusone A completely inhibited the planktonic growth of S. aureus at 250 and 500 µ g/mL; however, it did not have the same activity on E. coli. Derriobtusone A reduced the biomass and colony-forming unit (cfu) of S. aureus biofilm at concentrations of 250 and 500 µ g/mL. In various concentrations, it reduced the biofilm biomass of E. coli, and, in all concentrations, it weakly reduced the cfu. Derriobtusone A showed highly efficient antioxidant ability in scavenging DPPH radical and inhibiting ß -carotene oxidation. The compound showed no lethality to Artemia sp. nauplii. In conclusion, derriobtusone A may be an effective molecule against S. aureus and its biofilm, as well as a potential antioxidant compound with no toxicity.

Antimicrobial and antibiofilm action of Casbane Diterpene from Croton nepetaefolius against oral bacteria.

OBJECTIVE: The antibacterial activity of Casbane Diterpene (CD) was evaluated in vitro against Streptococcus oralis, S. mutans, S. salivarius, S. sobrinus, S. mitis and S. sanguinis. The viability of planktonic cells was analysed by susceptibility tests (MIC and MBC) and antibiofilm action was assayed. METHODS: The minimal inhibitory and bactericidal concentrations (MIC and MBC) of oral Streptococcus were evaluated through microdilution tests. To assay antibiofilm activity, biofilms were generated on 96-wells polystyrene plates under the presence of CD and quantified by a crystal violet technique and colonies forming units counting. RESULTS: The CD isolated from Croton nepetaefolius showed antimicrobial effect on planktonic forms and biofilms of oral pathogens, with MIC values of 62.5 µg/mL against Streptococcus oralis and values between 125 and 500 µg/mL against S. mutans, S. salivarius, S. sobrinus, S. mitis and S. sanguinis. CD showed an inhibitory effect on S. mutans biofilm formation at 250 µg/mL, and a decrease on viable cell of 94.28% compared to the normal biofilm growth. CONCLUSIONS: The compound CD can be considered as a promising molecule for the treatment against oral pathogens responsible for dental biofilm.

Casbane diterpene as a promising natural antimicrobial agent against biofilm-associated infections.

Croton nepetaefolius is a native plant from northeastern Brazil that belongs to the Euphorbiaceae family. The biological action of this plant has been extensively explored, being the secondary metabolites responsible for its properties alkaloids, diterpenes, and triterpenes. This study aimed to evaluate the ability of casbane diterpene (CD), isolated from the ethanolic extract of C. nepetaefolius, to inhibit microbial growth and biofilm formation of several clinical relevant species (bacteria and yeasts). It was found that CD possessed biocidal and biostatic activity against the majority of the species screened, with minimal active concentrations ranging between 125 and 500 µg/mL. In addition, it was observed that biofilm formation was inhibited even when the planktonic growth was not significantly affected. In conclusion, CD showed potential to be a natural tool for the treatment of diseases caused by different infectious microorganisms.