ABSTRACT
Avaliou-se o potencial terapêutico frente à cárie dentária da tintura hidroalcoólica (20%) de Bauhinia forficata Link (TBF). O estudo foi dividido em três artigos. O primeiro avaliou a constituição química mineral, pH, sólidos solúveis totais (°Brix) da TBF, suas concentrações mínimas inibitória (CMI) e microbicida (CMM) frente aos Streptococcus mutans (SM), o perfil de crescimento/morte (48h) desse microrganismo na presença da TBF, a citotoxicidade da tintura frente a fibroblastos orais (L929) na concentração máxima de 2,33mg/mL, e a ação antibacteriana da TBF, comparada aos controles (clorexidina 0,12% - CHX, dentre eles), sobre um biofilme de SM formado (48h) em blocos de esmalte bovino (n=48), após 3 dias de tratamento diário (1 min). No segundo artigo, investigou-se a CMI e CMM frente ao SM e Streptococcus sanguinis, Candida albicans, Fusubacterium nucleatum, Lactobacillus casei, Prevotella nigrescens, Bifidobacterium dentium; e a ação antimicrobiana da TBF sobre um biofilme de pool salivar de adultos (1mL, n=3, foram misturados e 20µL semeados para formação do biofilme) em membranas de celulose (n=4, 48h) tratadas em um único momento (membranas submersas por 1 min). A ação antimicrobiana foi quantificada pela contagem de Streptococcus spp. (S) e microrganismos totais (MT). No terceiro artigo, observou-se os conteúdos fenólicos da TBF, perfil de crescimento/morte dos microrganismos testados na presença da tintura, a citotoxicidade da TBF (23,21 e 200 mg/mL) até 5 min de contato com fibroblastos orais e seus efeitos: antimicrobiano frente um biofilme formado de pool salivar de crianças (contagem de S e MT e avaliação da altura do biofilme por microscopia confocal) e sobre a perda mineral do esmalte dentário (n=60; % de perda de microdureza-%PMD) após tratamentos diários (TBF e controles) por 7 dias (50µL/1min). Os dados foram analisados descritivamente e inferencialmente (p<0,05). No primeiro artigo, detectou-se cálcio, fósforo e magnésio na composição da tintura, pH=5,8 e ºBrix=17. A TBF apresentou CIM e CBM de 1.11mg/mL e promoveu redução óptica (0,32 nm) do SM após 48h (p<0,05). A tintura obteve baixos valores de morte celular (fibroblastos orais) (p<0,05) e em 2.32 mg/mL reduziu os níveis de SM do biofilme similar à CHX (p>0,05), sendo efetiva em sua redução (p<0,05). No segundo artigo, as CMI e CMM variaram de 1,31-23,2mg/mL e a TBF reduziu S e MT comparada ao não tratamento do biofilme (p<0.05), porém com ação inferior à CHX (p<0,05). No artigo três, a TBF apresentou grande quantidade de compostos fenólicos e ácidos clorogênicos. Na CMM de cada microrganismo gerou uma redução óptica após 48h; promoveu, na maior CMM (23,21mg/mL) baixa citotoxicidade (3.20% de células mortas em 5 min) e reduziu S e MT comparada ao controle de crescimento (CC; p<0,05), mas com ação inferior à CHX (p<0,05). Além disso, não alterou a altura da biomassa formada sobre os blocos (p>0,05) e permitiu menor %PMD comparada ao CC (p<0,05). Portanto, conclui-se que a TBF é uma substância com potencial terapêutico frente à cárie dentária, por apresentar ação antimicrobiana frente a diferentes biofilmes dentários com uso tópico oral seguro. (AU)
It was evaluated the therapeutic potential against dental caries of a hydroalcoholic tincture (20%) of Bauhinia forficata Link (TBF). The study was divided into three articles. The first one evaluated the mineral chemical composition, pH, total soluble solids (°Brix) of TBF. The TBF minimum inhibitory (MIC) and microbicide (MMC) concentrations against Streptococcus mutans (SM); the growth / death profile of SM (48h) with TBF; and the cytotoxicity of the tincture in oral fibroblasts (L929) at a maximum concentration of 2.33 mg/mL were also evaluated. The antibacterial action of TBF, compared to controls (chlorhexidine 0.12% - CHX, among them), on a SM biofilm formed (48h) on bovine enamel blocks (n=48) after 3 days of daily treatment (1 min) was observed. In the second article, MIC and MMC against SM and Streptococcus sanguinis, Candida albicans, Fusubacterium nucleatum, Lactobacillus casei, Prevotella nigrescens, Bifidobacterium dentium were investigated. The antimicrobial action of TBF was also tested against a salivary pool biofilm of adults (1mL, n=3, were mixed and 20µL seeded for biofilm formation) on cellulose membranes (n=4, 48h) treated in a single moment (membranes submerged during 1 min). The antimicrobial action was quantified by counting Streptococcus spp. (S) and total microorganisms (TM). In the third article, the phenolic contents of the TBF; growth/death profile of the tested microorganisms in the presence of the tincture; and the cytotoxicity (23.21 and 200 mg/mL) of TBF up to 5 min of contact with oral fibroblasts were evaluated. The antimicrobial effect of TBF against a biofilm formed from a salivary pool of children (counting S and TM and evaluation of biofilm height by confocal microscopy) on enamel bovine blocks of teeth and the tincture effect against the dental enamel mineral loss (n=60; % microhardness loss- %MHL) after daily treatments (TBF and controls) of the biofilm until 7 days (50µL/1 min) were observed. Data were analyzed descriptively and inferentially (p<0.05). In the first article, calcium, phosphorus and magnesium were detected. The, pH was 5.8 and ºBrix=17. TBF showed MIC and MMC of 1.11mg/mL and was able to decrease the optics density (0.32 nm) of SM after 48 hours (p<0.05). Low values of cell (oral fibroblasts) death (p<0.05) was obtained in the presence of TBF and it (2.32 mg/mL) reduced SM levels of the biofilm similarly to CHX (p>0.05), being effective in SM reducing (p<0.05). In the second article, the MIC and MMC ranged from 1.31-23.2mg/mL and TBF reduced S and MT compared to non-biofilm treatment (p<0.05), but with an antimicrobial action lower than CHX (p <0.05). In the third article, the TBF presented large amount of phenolic compounds and chlorogenic acids. The TBF at the MMC of each microorganism reached an microbial optical reduction after 48h. Even with the highest TBF MMC (23. 21mg/mL), a low cytotoxicity (3.20% of dead cells at 5 min) was observed. And the tincture (23. 21mg/mL) reduced S and TM compared to growth control (CC, p<0.05), but lower than CHX (p<0.05). In addition, TBF did not change the height of the biomass formed on the blocks (p>0.05) and allowed lower %MHL compared to CC (p<0.05). Therefore, it is concluded that TBF is a substance with therapeutic potential against dental caries, since it presents antimicrobial action against different dental biofilms with safe oral topical use. (AU)
Subject(s)
Bauhinia/chemistry , Dental Caries/drug therapy , Dental Plaque/prevention & control , Anti-Infective Agents/therapeutic use , In Vitro Techniques , Hydroalcoholic Solution , Bauhinia/toxicityABSTRACT
BACKGROUND: Previous experiments have shown that a decoction of Bauhinia forficata leaves reduces the changes in carbohydrate and protein metabolism that occur in rats with streptozotocin-induced diabetes. In the present investigation, the serum activities of enzymes known to be reliable toxicity markers were monitored in normal and streptozotocin-diabetic rats to discover whether the use of B. forficata decoction has toxic effects on liver, muscle or pancreas tissue or on renal microcirculation. METHODS: An experimental group of normal and streptozotocin-diabetic rats received an aqueous decoction of fresh B. forficata leaves (150 g/L) by mouth for 33 days while a control group of normal and diabetic rats received water for the same length of time. The serum activity of the toxicity markers lactate dehydrogenase, creatine kinase, amylase, angiotensin-converting enzyme and bilirubin were assayed before receiving B. forficata decoction and on day 19 and 33 of treatment. RESULTS: The toxicity markers in normal and diabetic rats were not altered by the diabetes itself nor by treatment with decoction. Whether or not they received B. forficata decoction the normal rats showed a significant increase in serum amylase activity during the experimental period while there was a tendency for the diabetic rats, both treated and untreated with decoction, to have lower serum amylase activities than the normal rats. CONCLUSIONS: Administration of an aqueous decoction of B. forficata is a potential treatment for diabetes and does not produce toxic effects measurable with the enzyme markers used in our study.
Subject(s)
Amylases/metabolism , Bauhinia/toxicity , Diabetes Mellitus, Experimental/therapy , Phytotherapy , Plant Leaves/toxicity , Animals , Bilirubin/metabolism , Creatine Kinase/metabolism , Diabetes Mellitus, Experimental/chemically induced , Diabetes Mellitus, Experimental/metabolism , L-Lactate Dehydrogenase/metabolism , Male , Peptidyl-Dipeptidase A/metabolism , Rats , Rats, Wistar , Streptozocin , Toxicity Tests, ChronicABSTRACT
Seeds of Bauhinia forficata species were submited to biochemical characterization concerning fatty acids analysis, protein fractionization, and hemaglutinanting activity. The seed elementary analysis showed a high protein and lipids contents with 21.24% and 19.45% respectively. The more abundant fatty acid was linoleic acid with 46.47% of the lipidic fraction. With the exception of prolamins, the different proteic fractions (albumin, globulins, acid and basic glutelins) showed hemaglutinanting activity against rabbit red cells no treated and treated with proteolitic enzymes. The fraction acid glutelin showed the higher specific hemaglutinanting activity (1072.25 UH/mg P) against rabbit blood pre-treated with trypsin. Glutamin (16.20%) and Valin (11.07%) were the more abundant amino acids in the seeds. Therefore, B. forficata represent a possible optional source of food because exhibit a high energetic values.