tannins and Hemorrhage

In humans, excessive bleeding during civilian accidents, and surgery account for 40% of the mortality worldwide. Hence, the development of biocompatible hemostatic materials useful for rapid hemorrhage control has become a fundamental research problem in the biomedicine community. In this study, we prepared biocompatible gelatin-tannic acid-κ-carrageenan (GTC) microparticles using a facile Tween 80 stabilized water-in-oil (W/O) emulsion method for rapid hemostasis. The formation of GTC microparticles occurs via polyelectrolyte interactions between gelatin and k-carrageenan as well as hydrogen bonding from tannic acid. In addition, the GTC microparticles formulated in our study showed high water adsorption ability with a low volume-swelling ratio for a particle size of 46 μm. In addition, the GTC microparticles displayed >80% biocompatibility in NIH 3T3 cells and <5% hemocompatibility in hemolysis ratio tests. Notably, the GTC microparticles induced rapid blood clotting in 50 s and blood loss of approximately 46 mg in the femoral artery of BALB/c female mice with a 100% survival rate that was significantly better than the control group (blood clot time:250 s; blood loss: 259 mg). Thus, the findings from our study collectively suggest that GTC microparticles may play a promising clinical role in medical applications to tackle hemorrhage control.

Topics: Animals; Carrageenan; Cross-Linking Reagents; Female; Gelatin; Hemorrhage; Hemostatics; Mice; Mice, Inbred BALB C; NIH 3T3 Cells; Polyelectrolytes; Tannins

Effective tissue hemostasis in periapical surgical site is important in the procedures. Plants with large amount of tannins may act as a local hemostatic agent. We aimed to compare the hemostatic effect of the extract of Quercus persica with one of the common hemostatic material used in periapical surgery. Six standardized bone holes were prepared in the calvaria of 5 Burgundy rabbits. Two hemostatic medicaments were tested for their hemostatic effect and were compared with control defects: Group 1, cotton pellet soaked in 15.5% ferric sulfate solution; Group 2, cotton pellet soaked in pure ethanolic extract of Q. persica. Bleeding score between the groups was compared. The ferric sulfate group exhibited significantly less bleeding than the other 2 groups. Q. persica was found to cause more hemostasis than the control group at 4 and 5 minutes but there were no significant differences between normal saline and Q. persica extract in bleeding control.

Topics: Animals; Ferric Compounds; Hemorrhage; Hemostatics; Plant Extracts; Quercus; Rabbits; Skull; Tannins

The plant species Connarus favosus is used in folk medicine in the west of Pará state, Brazil, to treat snakebites.. To investigate the potential of the aqueous extract of Connarus favosus (AECf) to inhibit hemorrhagic and phospholipase A2 activities induced by Bothrops atrox venom (BaV) and to determine the antioxidant and antimicrobial potentials of the extract.. AECf was analyzed phytochemically for phenolics (condensed tannins and hydrolyzable tannins) by colorimetry. Antioxidant activity was evaluated by quantitative assays using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and Fe(3+)/phenanthroline. Antimicrobial activity was evaluated by the minimal inhibitory concentration test, and cytotoxicity was evaluated using human fibroblast cells (MRC-5). Inhibition of BaV-induced hemorrhagic activity was assessed after oral administration of the extract using pre-treatment, post-treatment and combined (BA plus AECf) treatment protocols. Inhibition of indirect hemolysis caused by phospholipase A2 (PLA2) was investigated in vitro. Interaction between AECf and BaV was investigated by SDS-PAGE electrophoresis, Western blot (Wb) and zymography.. The phytochemical profile of AECf revealed ten secondary metabolite classes, and colorimetry showed high total phenolic and total (condensed and hydrolyzable) tannin content. AECf exhibited high antioxidant and antimicrobial potentials. The IC50 for the cytotoxic effect was 51.91 (46.86-57.50)µg/mL. Inhibition of BaV-induced hemorrhagic activity was significant in all the protocols, and inhibition of PLA2 activity was significant with the two highest concentrations. The BaV/AECf mixture produced the same bands as BaV by itself in SDS-PAGE and Wb although the bands were much fainter. Zymography confirmed the proteolytic activity of BaV, but when the venom was pre-incubated with AECf this activity was blocked.. AECf was effective in reducing BaV-induced hemorrhagic activity when administered by the same route as that used in folk medicine and exhibited antioxidant and antimicrobial potentials.

Topics: Animals; Anti-Bacterial Agents; Antioxidants; Antivenins; Bothrops; Brazil; Connaraceae; Crotalid Venoms; Female; Hemorrhage; Male; Medicine, Traditional; Mice; Phospholipases A2; Phytochemicals; Plant Extracts; Snake Bites; Tannins

Bellucia dichotoma Cogn. (Melastomataceae) is one of various plant species used in folk medicine in the west of the state of Pará, Brazil, to treat snake bites. Many studies have been carried out to evaluate the effectiveness of anti-snake bite plants, but few of these use the same preparation methods and doses as those traditionally used by the local populations. This study therefore compared inhibition of the main local effects of B. atrox venom (BaV) by aqueous extract of B. dichotoma (AEBd) administered according to traditional methods and pre-incubated with BaV). The concentrations of phenolic compounds (tannins and flavonoids) in AEBd were determined by colorimetric assays. The effectiveness of AEBd in inhibiting the hemorrhagic and edematogenic activities of BaV was evaluated in mice in four different experimental in vivo protocols: (1) pre-incubation (venom:extract, w/w); (2) pre-treatment (p.o.); (3) post-treatment (p.o.); and (4) AEBd (p.o.) in combination with Bothrops antivenom (BA) (i.v.). To assess in vitro inhibition of BaV phospholipase A₂ activity, the pre-incubation method or incorporation of AEBd or BA in agarose gels were used. The effect of AEBd on BaV was determined by SDS-PAGE, zymography and Western blot. Colorimetric assays revealed higher concentrations of (condensed and hydrolyzable) tannins than flavonoids in AEBd. Hemorrhagic activity was completely inhibited using the pre-incubation protocol. However, with pre-treatment there was no significant inhibition for the concentrations tested, and with the post-treatment only the 725 mg/kg dose of AEBd was able to inhibit 40.5% (p = 0.001) of the hemorrhagic activity of BaV. Phospholipase A₂ activity was only inhibited when AEBd was pre-incubated with BaV. BaV-induced edema was completely inhibited with pre-incubation (p < 0.05) and significantly reduced (p < 0.05) with pre- and post-treatment (p.o.) for the concentrations tested. The reduction in local edema was even greater when AEBd was administered in combination with BA. The SDS-PAGE profiles showed that several of the BaV protein (SDS-PAGE) and enzyme (zymography) bands were not detected when the venom was pre-incubated, and Western blot revealed that this was not caused by the AEBd enzymes observed in the zymogram. The "pseudo inhibition" observed after pre-incubation in this study may be due to the presence of tannins in the extract, which could act as chelating agents, removing metalloproteins and Ca²⁺ ions and thus inhibiti

Topics: Animals; Antivenins; Bothrops; Brazil; Crotalid Venoms; Edema; Enzyme Inhibitors; Ethnopharmacology; Group II Phospholipases A2; Hemorrhage; Hemostatics; Male; Medicine, Traditional; Melastomataceae; Mice; Neurotoxins; Plant Bark; Plant Extracts; Reptilian Proteins; Skin; Snake Bites; Tannins

Different fractions (I-V) of the methanolic leaf extracts of Annona senegalensis were assessed for their anti-snake venom activities. Fractions III neutralized lethal toxicity induced by Echis ocellatus venom and manifested the same potency as the crude extracts against the venom. The anti-snake venom activity of fraction III was clearly shown by the complete abrogation of venom-induced haemorrhage and the 75% record of surviving mice which were injected with a pre-incubate of venom and extract in the ratio 1:30 w/w after a 24 h. Also, fraction III exhibited a weak inhibitory effect on fibrinogen clotting activity of this venom. The key phytochemicals mediating the activity of this fraction are flavonoids and tannins. The detoxification of this venom by fraction III and the possible mode of action in the pathology of snake envenoming is discussed in this report.

Topics: Animals; Annona; Antivenins; Blood Coagulation; Fibrinogen; Flavonoids; Hemorrhage; Mice; Phytotherapy; Plant Extracts; Plant Leaves; Snake Bites; Tannins; Venoms; Viperidae

The aqueous extract prepared from Schizolobium parahyba (Sp) leaves, a native plant from Atlantic Forest (Brazil), was tested to analyse its ability to inhibit some biological and enzymatic activities induced by Bothrops alternatus (BaltCV) and Bothrops moojeni (BmooCV) snake venoms. Sp inhibited 100% of lethality, blood incoagulability, haemorrhagic and indirect haemolytic activities at a 1:10 ratio (venom/extract, w/w), as well as coagulant activity at a 1:5 ratio (venom/extract, w/w) induced by both venoms. BaltCV fibrinogenolytic activity was also neutralized by Sp at a 1:10 ratio, resulting in total protection of fibrinogen Bbeta chain and partial protection of Aalpha chain. Interaction tests have demonstrated that, at certain extract/proteins ratios, Sp precipitates proteins non-specifically suggesting the presence of tannins, which are very likely responsible for the excellent inhibiting effects of the analysed ophidian activities. Sp aqueous extract chromatography on Sephadex LH-20 was carried out aiming at the separation of these compounds that mask the obtained results. Thus, the fractionation of Sp resulted in three fractions: F1 (methanolic fraction); F2 (methanol:water fraction, 1:1 v/v); and F3 (aqueous fraction). These fractions were analysed for their ability to inhibit the BaltCV fibrinogenolytic activity. F1 inhibited 100% the venom fibrinogenolytic activity without presenting protein precipitation effect; F2 showed only partial inhibition of this venom activity. Finally, F3 did not inhibit fibrinogen proteolysis, but presented strong protein precipitating action. We conclude that Sp aqueous extract, together with tannins, also contains other compounds that can display specific inhibitory activity against snake venom toxins.

Topics: Animals; Anticoagulants; Antifibrinolytic Agents; Blood Coagulation; Bothrops; Chromatography, Gel; Fabaceae; Fibrinogen; Hemorrhage; Male; Mice; Phospholipase A2 Inhibitors; Phospholipases A2; Plant Extracts; Plant Leaves; Tannins; Viper Venoms

The use of plants as medicine has been referred to since ancient peoples, perhaps as early as Neanderthal man. Plants are a source of many biologically active products and nowadays they are of great interest to the pharmaceutical industry. The study of how people of different culture use plants in particular ways has led to the discovery of important new medicines. In this work, we verify the possible activity of Musa paradisiaca L. (Musaceae) against the toxicity of snake venoms. Musa paradisiaca, an important source of food in the world, has also been reported to be popularly used as an anti-venom. Interaction of Musa paradisiaca extract (MsE) with snake venom proteins has been examined in this study. Phospholipase A2 (PLA2), myotoxic and hemorrhagic activities, including lethality in mice, induced by crotalidae venoms were significantly inhibited when different amounts of MsE were mixed with these venoms before assays. On the other hand, mice that received MsE and venoms without previous mixture or by separated routes were not protected against venom toxicity. Partial chemical characterization of MsE showed the presence of polyphenols and tannins and they are known to non-specifically inactivate proteins. We suggest that these compounds can be responsible for the in vitro inhibition of the toxic effects of snake venoms. In conclusion, according to our results, using mice as experimental model, MsE does not show protection against the toxic effects of snake venoms in vivo, but if was very effective when the experiments were done in vitro.

Topics: Animals; Crotalid Venoms; Drug Evaluation, Preclinical; Flavonoids; Fruit; Hemorrhage; Male; Mice; Musa; Muscle, Skeletal; Neurotoxins; Phenols; Phospholipases A; Phospholipases A2; Plant Extracts; Plants, Medicinal; Polyphenols; Spectroscopy, Fourier Transform Infrared; Tannins

Topics: Acute Disease; Adrenocortical Hyperfunction; Adult; Alopecia; Animals; Antidotes; Asthenia; Colchicine; Coma; Diarrhea; Diuresis; Endometrium; Enzyme Inhibitors; Exchange Transfusion, Whole Blood; Fallopian Tubes; Feeding and Eating Disorders; Female; Hemorrhage; Humans; Leukocytosis; Mice; Mitosis; Myelin Sheath; Neuromuscular Junction; Polyneuropathies; Rats; Seizures; Tannins; Vitamin B Complex; Vomiting

Topics: Administration, Intravenous; Animals; Hemorrhage; Injections; Injections, Intravenous; Injections, Subcutaneous; Lagomorpha; Prothrombin Time; Rabbits; Rats; Research; Tannins

Topics: Blood Coagulation; Capillary Permeability; Hemorrhage; Humans; Permeability; Tannins