Antimicrobial potential of plant extracts from the Brazilian Cerrado.

Bacteria are related do different oral diseases, such as dental caries and periodontal disease. Therefore, the control or/and eradication of microorganisms and their by-products is primordial for the success of their treatment. An alternative for decrease bacterial load is the use of plant extracts used in popular medicine. The cytotoxicity and antimicrobial action of extracts of Cariniana rubra Gardiner ex Miers, Senna martiniana, Anadenanthera colubrina (Vell.) Brenan and Spiranthera odoratissima St. Hil. against strains of Streptococcus mutans, Enterococcus faecalis, Staphylococcus aureus, Escherichia coli, Aggregatibacter actinomyces- tencomitans and Candida albicans were investigated. Cytotoxicity was assessed at concentrations of 1, 10, 40, 80, 100 and 1000 µg/mL by means of the MTT test and compared to a control group with untreated cells. Those with acceptable cytotoxicity had the antimicrobial action measured by the XTT test. As a positive control, sodium hypochlorite was used. Cariniana rubra Gardiner ex Miers had the highest citototoxicity results while Spiranthera odoratissima St. Hil. had the best results, but all extracts showed acceptable cytotoxicity at different concentrations. The plant extracts showed higher activity against A. actinomycetencomitans: Anadenanthera columbrina (Vell.) Brenan (80.52%) at 40 µg/mL, Spiranthera odoratissima St. Hil (78.48%) in 1 µg/mL, Senna martiniana (73.28%) in the concentration of 40 µg/mL and Cariniana rubra Gardiner ex Miers (70.50%) in 10 µg/mL. All extracts analyzed showed acceptable cytotoxicity at different concentrations and were promising for inhibition of the pathogenic microorganisms studied.

Antimicrobial potential of plant extracts from the Brazilian Cerrado

Abstract Bacteria are related do different oral diseases, such as dental caries and periodontal disease. Therefore, the control or/and eradication of microorganisms and their by-products is primordial for the success of their treatment. An alternative for decrease bacterial load is the use of plant extracts used in popular medicine. The cytotoxicity and antimicrobial action of extracts of Cariniana rubra Gardiner ex Miers, Senna martiniana, Anadenanthera colubrina (Vell.) Brenan and Spiranthera odoratissima St. Hil. against strains of Streptococcus mutans, Enterococcus faecalis, Staphylococcus aureus, Escherichia coli, Aggregatibacter actinomyces- tencomitans and Candida albicans were investigated. Cytotoxicity was assessed at concentrations of 1, 10, 40, 80, 100 and 1000 μg/mL by means of the MTT test and compared to a control group with untreated cells. Those with acceptable cytotoxicity had the antimicrobial action measured by the XTT test. As a positive control, sodium hypochlorite was used. Cariniana rubra Gardiner ex Miers had the highest citototoxicity results while Spiranthera odoratissima St. Hil. had the best results, but all extracts showed acceptable cytotoxicity at different concentrations. The plant extracts showed higher activity against A. actinomycetencomitans: Anadenanthera columbrina (Vell.) Brenan (80.52%) at 40 μg/mL, Spiranthera odoratissima St. Hil (78.48%) in 1 μg/mL, Senna martiniana (73.28%) in the concentration of 40 μg/mL and Cariniana rubra Gardiner ex Miers (70.50%) in 10 μg/mL. All extracts analyzed showed acceptable cytotoxicity at different concentrations and were promising for inhibition of the pathogenic microorganisms studied.

Resumo Bactérias estão relacionadas a diferentes doenças bucais, como a cárie dentária e a doença periodontal. Assim, o controle e/ou erradicação de microrganismos e seus subprodutos é primordial para o sucesso dos tratamentos. Uma alternativa para diminuir a carga bacteriana é a utilização de extratos vegetais utilizados na medicina popular. A citotoxicidade e ação antimicrobiana de extratos de Cariniana rubra Gardinerex Miers, Senna martiniana H.S. Irwin & Barneby, Anadenanthera colubrina (Vell.) Brenan e Spiranthera odoratissima St. Hil. contra cepas de Streptococcus mutans, Enterococcusfaecalis, Staphylococcus aureus, Escherichia coli, Agartibacter actinomycetencomitans e Candida albicans foram investigados. A citotoxicidade foi avaliada nas concentrações de 1, 10, 40, 80, 100 e 1000 μg/mL por meio do teste MTT. Aqueles com citotoxicidade aceitável tiveram a ação antimicrobiana medida pelo teste XTT. Cariniana rubra Gardinerex Miers apresentou os maiores resultados de citototoxicidade, enquanto Spiranthera odoratissima St. Hil. obteve os melhores resultados, mas todos os extratos apresentaram citotoxicidade aceitável em diferentes concentrações. Os extratos vegetais apresentaram maior atividade contra A. actinomycetencomitans: Anadenanthera columbrina (Vell.) Brenan (80,52%) a 40 μg/mL, Spiranthera odoratissima St. Hil (78,48%) em 1 μg/mL, Senna martiniana H.S. Irwin & Barneby (73,28%) na concentração de 40 μg/mL e Cariniana rubra Gardinerex Miers (70,50%) em 10 μg/mL. Todos os extratos analisados apresentaram citotoxicidade aceitável em diferentes concentrações e foram promissores na inibição dos microrganismos patogênicos estudados.

Evaluation of Myrcia bella in murine osteosarcoma cells: Effect of the extract and enriched fractions of tannins and flavonoids.

Myrcia bella Cambess (Myrtaceae) is an important and common plant, native to the Brazilian Cerrado, with cytotoxicity, antimicrobial, and antidiabetic properties. Therefore, the effects of crude hydroalcoholic extract (CE) and fractions of ellagitannins (ELT) and flavonoids (FV) from Myrcia bella leaves were evaluated in a UMR-106 murine osteosarcoma cells and MC3T3 (normal cell). Cell viability and migration, production of reactive oxygen species (ROS) and matrix metalloproteinase (MMP) -2 and -9 activities were evaluated. In general, CE (80 µg/mL), ELT (160 µg/mL) and FV (64 µg/mL) reduced cell viability (p < 0.05). FV (64 µg/mL) was more effective in inhibition of cell migration, ROS production, and MMP-2 activity when compared to CE and ELT. Myrcia bella a rich source of phenolic compounds and its fraction of flavonoids have cytotoxic effects on osteosarcoma cells, preserving the viability of normal osteoblasts. Due to its antioxidant capacity, flavonoid may be a new therapeutic strategy for cancer.

Antimicrobial action, cytotoxicity, calcium ion release, and pH variation of a calcium hydroxide-based paste associated with Myracrodruon urundeuva Allemão extract.

The extract of Myracrodruon urundeuva Allemão (aroeira), as a vehicle, associated with calcium hydroxide (CH) paste was evaluated based on cell viability, antimicrobial action, calcium ion release, and pH variation. Calcium hydroxide with propylene glycol was used as control. The pH variation was measured at 3, 24, 72, 168, 140, 360, and 720 h and calcium ion release was measured on days 7, 15, and 30. Cell viability was assessed with NIH/3T3 cells using MTT and crystal violet assays, after 24, 48, and 72 h. Antibacterial activity was determined by the disc diffusion method, while microbial reduction (Enterococcus faecalis) was evaluated using the time-kill test. The CH paste formulated with aroeira showed antibacterial activity against Enterococcus faecalis and further did not interfere with pH, calcium ion release, or cell viability; moreover, the formulation had antimicrobial activity and could serve as a vehicle for CH paste.

Effects of Qualea grandiflora Extract on the Expression of MMP-14 and HIF-1α in Cultured Fibroblasts and Preosteoblasts

Abstract Background: Qualea grandiflora (QG) (Vochysiaceae), also known as "pau-ferro", "pau-terra" or "pau-de-tucano", is a very common deciduous tree in the Brazilian Cerrado used in traditional medicine to treat inflammations, ulcers, diarrhea, and infections. There are reports in the scientific literature that demonstrate the medicinal effects of the bark and leaf of the QG. However, studies involving this plant are rather imited. Aim of the study: To perform the phytochemical analysis of the QG hydroalcoholic extract (HAE) of leaves, and to investigate it effects on fibroblast and preosteoblasts. Methods: Phytochemical analysis was done by HPLC-DAD. Murine NIH/3T3 fibroblasts and MC3T3-E1 preosteoblasts cell lines (ATCC) were used for the experiments. Cell viability was assessed by the MTT colorimetric assay and the expression of MMP-14 and HIF-1α by immunofluorescence. Results and conclusion: The following compounds were identified by HPLC-DAD, such as quinic acid, ethyl galate, ellagic acid derivatives as O-methylellagic acid O-galloyl, O-methylellagic acid O-deoxyhexoside, galloyl derivatives, flavonol glycoside as kaempferol-O-deoxyhexoside, quercetin-O-deoxyhexoside, myricetin-O-deoxyhexoside and the pentacyclic triterpene arjunglucoside. Cell viability results demonstrated no cytotoxic effects in the studied concentrations. We found in QG HAE some compounds with therapeutic properties that can increase the expression of MMP-14 and HIF-1α, in fibroblasts and preosteoblasts. These data suggest that QG HAE has an action on these two molecules widely involved in physiological conditions, such as collagen remodeling, bone development and growth and pathological processes as HIF signaling in cancer metastasis.

Updating the role of matrix metalloproteinases in mineralized tissue and related diseases.

Bone development and healing processes involve a complex cascade of biological events requiring well-orchestrated synergism with bone cells, growth factors, and other trophic signaling molecules and cellular structures. Beyond health processes, MMPs play several key roles in the installation of heart and blood vessel related diseases and cancer, ranging from accelerating metastatic cells to ectopic vascular mineralization by smooth muscle cells in complementary manner. The tissue inhibitors of MMPs (TIMPs) have an important role in controlling proteolysis. Paired with the post-transcriptional efficiency of specific miRNAs, they modulate MMP performance. If druggable, these molecules are suggested to be a platform for development of "smart" medications and further clinical trials. Thus, considering the pleiotropic effect of MMPs on mammals, the purpose of this review is to update the role of those multifaceted proteases in mineralized tissues in health, such as bone, and pathophysiological disorders, such as ectopic vascular calcification and cancer.

Effect of hydroalcoholic extract of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability and activity of microcosm biofilm and on enamel demineralization.

OBJECTIVES: The aim of this study was to assess the effect of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves hydroalcoholic extracts on viability and metabolism of a microcosm biofilm and on enamel demineralization prevention. METHODOLOGY: Microcosm biofilm was produced on bovine enamel using inoculum from pooled human saliva mixed with McBain saliva, under 0.2% sucrose exposure, for 14 days. The biofilm was daily-treated with the extracts for 1 min. At the end, it was analyzed with respect to viability by fluorescence, CFU counting and extracellular polysaccharides (phenol-sulphuric acid colorimetric assay) and lactic acid (enzymatic assay) production. The demineralization was measured by TMR. The data were compared using ANOVA or Kruskal-Wallis (p<0.05). RESULTS: M. urundeuva All. at 100, 10 and 0.1 µg/mL and Q. grandiflora Mart. at 100 and 0.1 µg/mL reduced biofilm viability similarly to positive control (chlorhexidine) and significantly more than the negative-vehicle control (35% ethanol). M. urundeuva at 1000, 100 and 0.1 µg/mL were able to reduce both lactobacilli and mutans streptococci CFU counting, while Q. grandiflora (1000 and 1.0 µg/mL) significantly reduced mutans streptococci CFU counting. On the other hand, the natural extracts were unable to significantly reduce extracellular polysaccharides and lactic acid productions neither the development of enamel carious lesions. CONCLUSIONS: The extracts showed antimicrobial properties on microcosm biofilm, however, they had no effect on biofilm metabolism and caries protection.

Low level laser therapy modulates viability, alkaline phosphatase and matrix metalloproteinase-2 activities of osteoblasts.

Low level laser therapy (LLLT) has been shown to stimulate bone cell metabolism but their impact on the matrix metalloproteinase (MMP) expression and activity is little explored. This study evaluated the influence of LLLT at two different wavelengths, red and infrared, on MC3T3-E1 preosteoblast viability, alkaline phosphatase (ALP) and MMP-2 and -9 activities. To accomplish this, MC3T3-E1 cells were irradiated with a punctual application of either red (660nm; InGaAIP active medium) or infrared (780nm; GaAlAs active medium) lasers both at a potency of 20mW, energy dose of 0.08 or 0.16J, and energy density of 1.9J/cm2 or 3.8J/cm2, respectively. The control group received no irradiation. Cellular viability, ALP and MMP-2 and -9 activities were assessed by MTT assay, enzymatic activity and zymography, respectively, at 24, 48 and 72h. The treatment of cells with both red and infrared lasers significantly increased the cellular viability compared to the non-irradiated control group at 24 and 48h. The ALP activity was also up modulated in infrared groups at 24 and 72h, depending on the energy densities. In addition, the irradiation with red laser at the energy density of 1.9J/cm2 promoted an enhancement of MMP-2 activity at 48 and 72h. However, no differences were observed for the MMP-9 activity. In conclusion, when used at these specific parameters, LLL modulates both preosteoblast viability and differentiation highlighted by the increased ALP and MMP-2 activities induced by irradiation.

Influence of iron on modulation of the antioxidant system in rat brains exposed to lead.

The objective of this study was to evaluate markers of oxidative stress in the brains of rats exposed to lead acetate (Pb(C2 H3 O2 )2 ), either associated or not associated with ferrous sulfate (FeSO4 ). A total of 36 weaning rats (Rattus norvegicus) were divided into 6 groups of six animals and exposed to lead acetate for six weeks. In the control group (control), the animals received deionized water. The Pb260 and Pb260 + Fe received 260 µM lead acetate, and the Pb1050 and Pb1050 + Fe received 1050 µM lead acetate. The Pb260 + Fe and Pb1050 + Fe were supplemented with 20 mg of ferrous sulfate/Kg body weight every 2 days. Group Fe received deionized water and ferrous sulfate. The rat brains were collected to analyze the enzymatic activity of catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and the concentration of reduced glutathione (GSH), lipid peroxidation (TBARS), and total antioxidant substance (TAS) (DPPH• technique). The activity of SOD and GPx in the experimental groups decreased compared to the control, together with the concentration of GSH (p < 0.05). For CAT analysis, SOD tended to increase in concentration in the experimental groups without a concomitant exposure to FeSO4 , whereas GPx showed a slight tendency to increase in activity compared to the control. For TAS-DPPH• , there was a decrease in the experimental groups (p < 0.05). According to the results, SOD, GPx, and GSH were affected by lead acetate and exposure to ferrous sulfate changed this dynamic. However, further studies are needed to verify whether ferrous sulfate acts as a protectant against the toxic effects of lead. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 813-822, 2017.

The Dynamics of Subcutaneous Tissue Response to Microorganisms Associated with the Extract of Araçá (Psidium cattleianum): An Edemogenic and Microscopic Analysis / A Dinâmica da Resposta do Tecido Subcutâneo para Associação de Microorganismos com o Extrato de Araçá (Psidium cattleianum): Uma Análise Microscópica e Edemogênica

Objective: To evaluate in vivo tissue reaction to the extract of araçá (Psidium cattleianum) associated with inactivated microorganisms. Material and Methods: A 0.1 mL suspension was used containing Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Enterococcus faecalis, Peptostreptococcus micros, and Porphyromonas endodontalis, which were inactivated by heat and mixed into a 1.0 mL saline (control group), an aqueous solution, or a hydroalcoholic extract of araçá. Eighteen male rats (Rattus norvegiccus) under general anesthesia received 0.2 mL of 1% intravenous Evans blue. Thirty minutes later, 0.1 mL of one of the associations was injected into the animals' dorsal region. The animals were euthanized after 3 and 6 hours, and the materials obtained were placed in formamide for 72 hours then analyzed in a spectrophotometer (λ=630 ηm). For the morphological analysis, 30 rats received polyethylene tubes implants with the extracts or the saline with the associations in the dorsal region and euthanized after 7 and 30 days to be analyzed according to an inflammation cell score. Results: No significant difference (p>0.05) was observed in the edema among groups. The optical microscopy results showed a repair in the 30-day-period, which was higher when compared to the 7-day-period (p< 0.0001). Nevertheless, in the 7-day-period, the hydroalcoholic extract presented a significant response compared to the aqueous extract (p=0.05) and a trend for better results than the control group. Conclusion: The aqueous and hydroalcoholic araçá extracts associated with inactivated microorganisms showed similar responses to control, indicating no interference on the toxic effects of the bacterial components in tissue repair. (AU)

Objetivo: Avaliar in vivo a reação tecidual do extrato de araçá (Psidium cattleianum) associado com microorganismos inativados. Material e Métodos: Uma suspensão de 0.1mL foi usada contendo Porphyromonas gingivalis, Prevotella intermedia, Fusobacterium nucleatum, Enterococcus faecalis, Peptostreptococcus micros e Porphyromonas endodontalis dos quais foram inativos por aquecimento e misturados a 1,0 mL de soro fisiológico (grupo controle), uma solução aquosa ou hidroalcoólica de araçá. Dezoito ratos machos (Rattus norvegiccus) sob anestesia geral receberam 0,2mL de Azul de Evans a 1% intravenoso. Após trinta minutos, 0,1mL de um dos extratos (associado com microorganismos inativos) foi injetado nos animais na região dorsal. Os animais foram eutanasiados após 3 e 6 horas, e os materiais obtidos colocados em formamida por 72 horas para análise em espectrofotômetro (λ=630 ηm). Para análise morfológica, 30 ratos receberam implante subcutâneo de tubo de polietileno com as associações na região dorsal, eutanasiados após 7 e 30 dias para serem analisados de acordo com um escore de células inflamatórias. Resultados: Não houve diferença significativa (p>0,05) no edema entre os grupos. Os resultados obtidos em microscópio óptico apontaram reparo em 30 dias superior ao de 7 dias (p< 0,0001). No período de 7 dias a solução hidroalcoólica apresentou resposta superior a solução aquosa (p=0,05) e uma tendência de melhor resultado que o controle. Conclusão: A solução aquosa e hidroalcoólica de extrato de araçá associadas a microrganismos inativados apresentaram respostas biológicas semelhantes ao controle, indicando que não há interferência sobre os efeitos tóxicos advindos dos componentes bacterianos, no sentido de favorecer o reparo(AU)

"Aroeira" (Myracrodruon urundeuva) methanol extract: the relationship between chemical compounds and cellular effects.

CONTEXT: "Aroeira" [Myracrodruon urundeuva Allemão (Anacardiaceae)] is a tree whose leaves have been studied for therapeutic purposes in medicine and dentistry. OBJECTIVE: The study chemically identifies the leaf extract of aroeira and determines its effect on human gingival fibroblasts. MATERIALS AND METHODS: An 80% methanol leave extract was obtained by maceration and chemically identified through flow-injection analysis-electrospray ionization-ion trap-tandem mass spectrometry (FIA-ESI-IT-MSn). Cytotoxicity of the aroeira's methanol extract was evaluated in lineage of fibroblasts. Adherent cells were treated with different concentrations of aroeira's methanol extract in the medium: 0.1, 1, 10, 100 and 1000 µg/mL. Control cells were cultivated in the medium only. Analyses were done at 24, 48, 72 and 96 h of culture by neutral red assay; and at 24, 48 and 96 h by crystal violet assay. RESULTS: FIA-ESI-IT-MS analysis determined the presence of compounds, for the first time in the species: quercetin-O-glucuronide and quercetin-O-deoxyhexose-O-glucose in the extract. On one hand, neutral red and crystal violet assay showed a reduction (to 50% up until 100%) of cellular viability of groups of 100 and 1000 µg/mL compared with control at 96 h (p < 0.05). On the other hand, lower concentrations (0.1; 1 and 10 µg/mL) of the extract were similar to that of the control at 96 h (p < 0.05), in general. CONCLUSIONS: In view of the results, we can conclude that the extract of aroeira presents tannins and flavonoids. Furthermore, the extract is capable of modulating the viability of human gingival fibroblasts according to its concentration.

Viability of Human Gingival Fibroblast (FGH) Treated with Ethanolic "Aroeira" Extract (Myracrodruon urundeuva Allemão)

ABSTRACT The aim of this study was to evaluate the effect of ethanolic "aroeira" (Myracrodruon urundeuva) extract on the viability of human gingival fibroblast. For this, fibroblasts (2x103 cells/well) were plated in a 96-well plate and incubated for 24 h; the medium (Eagle's medium modified by Dulbecco - DMEM) supplemented with 10% fetal bovine serum was replaced by DMEM with different ethanolic extract concentration (0, 0.1, 1, 10, 100, and 1000μg / mL). The fibroblast viability was analyzed after 48,72, and 96 h by the neutral red capture test and violet crystal. The "aroeira" extract, at high concentrations (100 and 1000 µg/mL) caused decrease in both cellular viability tests (p<0.05). However, dilutions between 0.1 and 10 µg/mL did not affect the viability of the cells. It was concluded that "aroeira" extract was able to change the gingival fibroblast viability, and this effect was concentration dependent.

The use of low-level laser therapy in the prevention and treatment of chemotherapy-induced oral mucositis

The management of oral mucositis is fundamental in maintaining the quality of life of cancer patients as well as the feasibility and effectiveness of non-surgical treatments of cancer. A literature review was carried out in order to update information on the use of low-level laser therapy for prevention and treatment of induced oral mucositis in patients undergoing chemotherapy or conditioning for hematopoietic cell transplantation. Lasertherapy is a simple, non-invasive and atraumatic technique which has shown good results in the prevention of oral mucositis through basic research and controlled clinical trials. Although the precise molecular mechanisms of low-level laser action have not been elucidated, in face of its proven effectiveness in the management of oral mucositis and the absence of side effects, its use should be continued and even encouraged while the results from new studies defining the optimal parameters for laser application are awaited.