Highly resilient, biocompatible, and antibacterial carbon nanotube/hydroxybutyl chitosan sponge dressing for rapid and effective hemostasis.

Uncontrolled hemorrhage is the leading cause of trauma death. The development of safe and efficient hemostatic agents that can rapidly and effectively control bleeding is of great significance to rescue the injured. However, the mechanical, absorptive, and antibacterial properties of conventional two-dimensional hemostatic agents are not satisfactory. Herein, a series of effective three-dimensional hemostatic dressings (JWCNT/HBC sponges) are developed by chemical modification of joint-welded carbon nanotube (JWCNT) sponges with hydroxybutyl chitosan (HBC) for hemorrhage hemostasis. The JWCNT/HBC sponges exhibit high elasticity, porous structure, and suitable blood-absorption and blood-maintaining performance. Moreover, the introduction of HBC endows the JWCNT/HBC sponges with favorable blood compatibility and good antibacterial activity. The sponge treated with 0.5% HBC (JWCNT/0.5%HBC sponge) displays better antiseptic capability, faster blood clotting ability in vitro and shorter hemostasis time in vivo than the commercial gelatin sponge. The JWCNT/HBC sponges combine the advantages of JWCNT sponges and HBC in the adhesion and activation of platelets and red blood cells, thus becoming a good medical material for trauma hemostasis.

Antibacterial and Hemostatic Thiol-Modified Chitosan-Immobilized AgNPs Composite Sponges.

Wound bleeding and infection are two of the major threats to patients' lives, but developing safe materials with high hemostasis efficiency and antibacterial activity remains a major challenge. Silver nanoparticles (AgNPs) are suitable as antibacterial agents in the hemostatic process, but the application is hampered because of easy accumulation of toxicity. Herein, thiol-modified chitosan (TMC) was prepared by modifying with mercaptosuccinic acid and then was used to immobilize AgNPs to obtain composite sponges (TMC/AgNPs) for stemming the bleeding and preventing infection. TMC/AgNPs sponges had complex interlaced tubular porous structure with high porosity (99.42%), indicating high absorption. TMC had high immobilization efficiency for AgNPs-the release rate of AgNPs was 14.35% after 14 days-but the TMC/AgNPs sponge still had excellent antibacterial activity against Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. In vitro and in vivo experiments confirm that the TMC/AgNPs sponge had fast and efficient hemostatic performance in comparison with the PVF sponge, and its possible mechanism was the synergistic effect of high blood absorption capacity and the interaction between amino, sulfydryl, and blood cells. Furthermore, the TMC/AgNPs sponge can promote wound healing by preventing wound infection, while the PVF sponge cannot. More importantly, the sponges had good safety due to the immobilization of TMC for AgNPs.

Analysing the blood-stemming effect of Ankaferd Blood Stopper in medulla spinalis surgery

Background/aim: The aim of this study was to investigate the possible toxicity of the Ankaferd Blood Stopper (ABS) on the neural system. Materials and methods: Thirty Sprague Dawley rats were randomized into ABS (n: 15) and control (n: 15) groups. Following the anaesthetic induction, total laminectomy was performed to the lower thoracic, and upper lumbar areas in both groups and medulla spinalis was exposed. Two myelotomies were performed on the medulla spinalis. One millilitre ABS was applied to the incision site in the ABS group, and one millilitre 0.9% saline solution was applied in the control group. Rats were observed for 15 days regarding general behaviour, neurological signs, mobility, and signs of infection. Sixteen days later, all rats were decapitated under anaesthesia. Medulla spinalis was removed en bloc from all rats and was stained with Heamatoxylin & Eosin and luxol fast blue. Results: There was no significant difference between the ABS group and the control group regarding oedema, gliosis, the intensity of inflammatory cells, the presence of neuronal degeneration, neuron counts, and myelin degeneration. Conclusion: No clinical or histopathological evidence for the neurotoxic effect of the ABS was observed in the present study. Our findings might precipitate the use of ABS on human subjects regarding medulla spinalis surgery.

Fabricating antimicrobial peptide-immobilized starch sponges for hemorrhage control and antibacterial treatment.

It is important to control immediate hemorrhage and prevent infection simultaneously in the wound management. However, most of hemostatic materials are associated with low efficiency of hemostasis, poor biocompatibility and lack of antimicrobial properties. A kind of starch-based macroporous sponges (KR-Sps) immobilized covalently with antimicrobial peptide KR12 using highly efficient thiol-ene photo click reaction were developed. The physical properties of these sponges could be fine-tuned by varying the ratio of modified starch/HS-PEG-SH and the polymer concentration. The in vitro and vivo results demonstrated that KR-Sps induced thrombosis, shortened clotting time and reduced the blood loss at bleeding site. Besides, KR12 immobilized sponge exhibited inherent antimicrobial properties against Gram (+) and (-) bacteria and methicillin-resistant Staphylococcus aureus (MRSA), which could maintain at least 5 days. Therefore, KR-Sps were believed to be an excellent candidate as hemostatic and antimicrobial product for the intraoperative wound management.

Assessment of the anti-snakebite properties of extracts of Aniba fragrans Ducke (Lauraceae) used in folk medicine as complementary treatment in cases of envenomation by Bothrops atrox.

ETHNOPHARMACOLOGICAL RELEVANCE: Extracts of leaves and bark of Aniba fragrans are used as tea (decoction) to treat snakebites in communities in the Brazilian Amazon. The aqueous extract of the leaves of A. fragrans has been proven to be effective against Bothrops venom, but only when pre-incubated with the venom. This study sought to assess the potential of different types of extract of this species to inhibit the biological activities of Bothrops atrox venom (BaV) when used the same way as in folk medicine. The main classes of secondary metabolites and the concentrations of phenolics in the extracts were also determined. MATERIALS AND METHODS: Four types of extract of A. fragrans were prepared: aqueous extract of the leaf (AEL), aqueous extract of the bark (AEB), hydroalcoholic leaf extract (HLE) and extract of the residue from hydrodistillation of the leaf (ERHL). The phytochemical profiles of the aqueous extracts were determined using thin layer chromatography (TLC), and the concentrations of phenolics were measured by colorimetric assays. To investigate the potential of the extracts to inhibit the biological activities of BaV, in vitro tests for antiphospholipase and antifibrinolytic activities were performed. In vivo tests for antihemorrhagic and antidefibrinating activities were also carried out, as well as antimicrobial tests for activity against the main bacteria found in the oral cavity of snakes. Interaction between the extracts and the proteins in BaV was assessed by electrophoresis (SDS-PAGE) and Western blot (WB). The cytotoxicity of the extracts was assessed in a strain of MRC-5 human fibroblasts. RESULTS: Terpenoids, flavonoids and condensed and hydrolysable tannins were detected in all the extracts. Metabolites such as coumarins, fatty acids and alkaloids were present in some extracts but not in others, indicating different phytochemical profiles. Phenolics content varied between extracts, and there were more tannins in AEB and HLE. In the in vitro tests, the extracts inhibited the phospholipase and fibrinolytic activities of BaV in the two ratios of venom to extract used. HLE exhibited effective antimicrobial action as it inhibited growth of 11 of the 15 bacteria investigated, including Morganella morganii, the main bacteria described in the oral cavity of snakes. The extracts failed to inhibit the defibrinating activity of BaV, and only the Bothrops antivenom had a significant effect (96.1%) on this activity. BaV-induced hemorrhage was completely inhibited by AEL and AEB when the pre-incubation (venom:extract) protocol was used. When administered orally, as in folk medicine, both AEB and AEL produced significant inhibition of hemorrhagic activity (maximum inhibition 46.5% and 39.2%, respectively). SDS-PAGE and WB of the extracts pre-incubated with BaV showed that the main proteins in the venom had been precipitated by the extracts. None of the four extracts showed cytotoxic effects in the tests carried out with a human fibroblast cell line. CONCLUSION: In addition to being effective in reducing hemorrhage when administered orally, the extracts displayed a high antimicrobial potential against microorganisms involved in secondary infections at the site of the snakebite. Once the extracts have been tested in accordance with the appropriate regulations, this species could potentially be used to produce a phytomedicine for complementary treatment of the secondary infections due to bacteria that aggravate the local signs and symptoms after snakebite envenomation.

Bletilla striata: Medicinal uses, phytochemistry and pharmacological activities.

ETHNOPHARMACOLOGICAL RELEVANCE: Bletilla striata (Thunb.) Reichb. f. (Orchidaceae), also known as Hyacinth Orchid and Baiji (Simplified Chinese: ), not only has been widely used for the treatment of hematemesis, hemoptysis, and traumatic bleeding due to the efficacy of arresting bleeding with astringent action, but also has been applied topically to overcome ulcers, sores, swellings, and chapped skin due to the efficacy of dispersing swelling and promoting tissue regeneration. Additional medical applications include the treatment of tuberculosis, malignant ulcers, hemorrhoids, anthrax, eye diseases, and silicosis. AIM OF THIS REVIEW: This review aims to provide up-to-date information on the botanical characterization, medicinal uses, chemical constituents, pharmacological activities, and toxicity of B. striata. In addition, this paper also focuses on the possible exploitation of this plant for the treatment of different diseases, and uncovers opportunities for future research. MATERIALS AND METHODS: The relevant information on B. striata was gathered from worldwide accepted scientific databases via an electronic search (Google Scholar, Web of Science, ScienceDirect, ACS Publications, PubMed, Wiley Online Library, SciFinder, CNKI). Information was also obtained from The Plant List, Chinese pharmacopoeia, Chinese herbal classics books, PhD and MSc dissertations, etc. RESULTS: A comprehensive analysis of the literature obtained through the above-mentioned sources confirmed that the ethnomedical usages of B. striata have been recorded in Mongolia, Korea, Japan, and China. Phytochemical investigations revealed that the major chemical constituents of B. striata are polysaccharides, bibenzyls, phenanthrenes, triterpenoids and its saponins, steroids and its saponins, which also have been proven to be the main bioactive substances capable of exhibiting numerous pharmacological activities including wound healing, antiulcer, hemostasis, cytotoxicity, antimicrobial, anti-inflammation, anti-oxidation, immunomodulation, anti-fibrosis, antiaging, anti-allergy, and anti-itch. CONCLUSIONS: Preliminary investigations on pharmacological properties of B. striata have shown that B. striata is an outstanding astringent hemostatic medicinal, B. striata polysaccharides (BSP) as the major bioactive components not only capable of promoting wound healing, but also show good performance as a kind of promising natural biomaterial. More importantly, BSP are also reported to be excellent embolic material. However, further investigations need to be carried out to fully clarify its efficacy of dispersing swelling and promoting tissue regeneration. Moreover, this plant also needs a lot more investigations to clarify the pathways of absorption, distribution, metabolism and excretion, and to evaluate its long-term in vivo chronic toxicity before proceeding to the development of pharmaceutical formulation.

Evaluation of cytotoxicity of a new hemostatic agent Ankaferd Blood Stopper® using different assays.

The aim of this study is to investigate the effects of the Ankaferd Blood Stopper® (ABS), on cell viability, cytotoxicity, and erythrocyte numbers in in vitro cultured human blood cells. We studied the cytotoxic effects of the ABS using lactate dehydrogenase (LDH) assay, cell proliferation (WST-1) assay and hemolytic assay. The cytotoxicity increased when cells were treated with ABS dilutions of 5%, 12.5%, 25%, and 50% (p < 0.05). Moreover, treatment of the cells with the same concentrations significantly elevated the cell number at 24 and 48 h (p < 0.05). ABS causes a significant increase (p < 0.05) in the hemolytic activity on human erythrocytes and hemolytic activity increases with increase in ABS concentrations. The red blood cell aggregation and cell membrane disruption during the coagulation process lead to induction of hemolytic activity and increase of LDH level in cell culture medium. In addition, ABS has proliferative effects on human leukocytes. Based on these results, ABS can be used as an alternative blood stopping agent safely.

Cytotoxicity of a new hemostatic agent on human pulp fibroblasts in vitro

Objective: The objective of this study was to evaluate the cytotoxicity of the plant extract ankaferd blood stopper(ABS) in vitro.Study Design: ABS was eluted with fresh Dulbecco’s Modified Eagle’s Medium (DMEM) without serum for 72h, at 37°C. The cells treated with various dilutions of ABS were seeded into 96-well microplate at 104 /well intriplicates. Cells without treatment served as a control group. The number of viable cells after 48 h incubation wasdetermined by a modified 3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. Therelative viability of pulp cells was expressed as color intensity of the number in the experimental wells relative tothat of the control group. Absorbances were read at 570 nm on a microplate reader with a background subtractionat 620 nm.Result: The results showed that ABS was cytotoxic to human pulp fibroblasts by MTT assay.Conclusions: The influence of cytotoxicity to human pulp fibroblasts depended on concentration of ABS. Themore dilutions exhibited less cytotoxic characteristics compared to the more concentrated forms (AU)

Cytotoxicity of a new hemostatic agent on human pulp fibroblasts in vitro.

OBJECTIVE: The objective of this study was to evaluate the cytotoxicity of the plant extract ankaferd blood stopper (ABS) in vitro. STUDY DESIGN: ABS was eluted with fresh Dulbecco's Modified Eagle's Medium (DMEM) without serum for 72 h, at 37 °C. The cells treated with various dilutions of ABS were seeded into 96-well microplate at 104/well in triplicates. Cells without treatment served as a control group. The number of viable cells after 48 h incubation was determined by a modified 3-(4, 5-dimethyl-thiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) assay. The relative viability of pulp cells was expressed as color intensity of the number in the experimental wells relative to that of the control group. Absorbances were read at 570 nm on a microplate reader with a background subtraction at 620 nm. RESULT: The results showed that ABS was cytotoxic to human pulp fibroblasts by MTT assay. CONCLUSIONS: The influence of cytotoxicity to human pulp fibroblasts depended on concentration of ABS. The more dilutions exhibited less cytotoxic characteristics compared to the more concentrated forms.

Effects of hepatic artery chemoembolization using cisplatin-lipiodol suspension with gelatin sponge particles on swine liver.

PURPOSE: To define the effects of hepatic artery chemoembolization with cisplatin-lipiodol suspension and gelatin sponge particles on swine liver tissue and estimate the concentration of cisplatin that would have a minimal negative effect on normal liver parenchyma. MATERIALS AND METHODS: Twelve pigs were divided into four groups: group A was the control group in which hepatic arteries were embolized with lipiodol and gelatin sponge particle (n = 3); group B animals were embolized with 10 mg/mL cisplatin-lipiodol suspension plus gelatin sponge particle (n = 3), group C with 20 mg/mL cisplatin-lipiodol suspension plus gelatin sponge particle (n = 3), and group D with 30 mg/mL cisplatin-lipiodol suspension plus gelatin sponge particle (n = 3). Pigs were euthanized 1 week after embolization, and the resected livers were cut into 10-mm-thick sections. The livers and necrotic foci were contoured in each section, and the necrosis volume ratio was calculated. RESULTS: The necrosis volume ratios of the livers in groups A, B, C, and D were 0.832% +/- 0.334, 2.324% +/- 1.126, 8.056% +/- 3.276, and 11.82% +/- 4.921, respectively. Significant differences (P < .05) in necrosis volume ratio were found between groups A and C, groups A and D, groups B and C, and groups B and D; no significant difference was found between groups A and B. CONCLUSIONS: Hepatic artery chemoembolization with higher doses of cisplatin causes greater damage to liver tissue; 10 mg/mL cisplatin-lipiodol suspension causes minimal damage, similar to that without cisplatin, and is related to minimal negative changes in a swine model.

Preclinical safety of recombinant human thrombin.

Recombinant human thrombin (rhThrombin) is being developed as an alternative to thrombin products purified from pooled human or bovine plasma, which are currently marketed for topical hemostasis. Preclinical studies of rhThrombin were conducted prior to its evaluation as a topical adjunct to surgical hemostasis in clinical trials. No overt clinical pathology or signs were observed in cynomolgus monkeys following implantation of a gelatin sponge containing either rhThrombin or bovine thrombin to a surgical liver wound, and similar gross and microscopic wound healing characteristics were observed over an eight-week recovery period with either compound. Repeated subcutaneous injections of rhThrombin or bovine thrombin to cynomolgus monkeys produced no treatment-related effects. Whereas no monkeys demonstrated anti-rhThrombin antibody seroconversion, specific anti-bovine antibodies were detected in all tested monkeys exposed to bovine thrombin. Addition of rhThrombin or bovine thrombin to mouse fibroblast cells resulted in expected detachment and shape change. Topical application of rhThrombin to rabbits did not cause irritation to the eye, normal skin, or abraded skin. These studies showed that topical, subcutaneous, or implanted rhThrombin was minimally immunogenic, safe, and well tolerated in nonclinical models, and supported the clinical evaluation of rhThrombin in a variety of surgical settings.

Operations on the thoracic aorta and hypothermic circulatory arrest: is aprotinin safe?

INTRODUCTION: The safety of aprotinin, especially when used with profound hypothermic circulatory arrest, is still a matter of intense debate despite its presumed salutary effects on blood loss. Many investigators have reported toxic renal effects of high-dose aprotinin in such patients, but no prospective, randomized study has been conducted. To assess the potential detrimental effect of aprotinin on renal function and its putative reduction of blood loss, 50 patients undergoing thoracic aortic operations with the use of profound hypothermic circulatory arrest were randomly assigned to receive either low-dose aprotinin (1 x 10(6) kallikrein activation units) or placebo. METHODS: The specific renal tubular markers beta-2-microglobulin and beta-N-acetyl-D-glucosaminidase, as well as serum creatinine and blood urea nitrogen, creatinine clearance, sodium excretion, and potassium excretion, were measured to evaluate renal function preoperatively, immediately after the procedure, and 24 hours and 48 hours later. RESULTS: No statistically significant difference was found in any measured renal parameter between the two groups (analysis of variance). Renal dysfunction, defined as an elevation of serum creatinine early postoperatively (> or = 1.5 times the preoperative value), occurred in two patients who received aprotinin and in one patient in the control group. Temporary dialysis (hemodialysis or continuous venovenous hemofiltration) was needed in two patients in the aprotinin group versus one in the control group. Furthermore, patients treated with aprotinin had significantly less total postoperative blood loss (718 +/- 340 ml vs 920 +/- 387 ml, p = 0.04). The aprotinin recipients also had a significantly lower transfusion requirement (p < 0.05). CONCLUSION: This controlled trial of low-dose aprotinin in patients undergoing thoracic aortic operations using profound hypothermic circulatory arrest demonstrated no detectable deleterious effects on renal function; moreover, the use of aprotinin was associated with significantly lower need for transfusion.

[Determination of pharmacokinetic parameters of shandahuang xiaoyan zhixue capsules].

Based on residual accumulation and biophase availability principles and using animal death rate as index, determination has been carried out of the residual accumulation rate as well as the toxicokinetic and pharmacokinetic parameters at different times of the Shandahuang Xiaoyan Zhixue Capsules.

[Research on hemostatic constituents in carbonized Schizonepeta tenuifolia Brig].

It has been shown that the fatsoluble extract SeE from carbonized Schizonepeta tenuifolia has an obvious hemostatic action. In a given range of dose there is a significant linear correlation between the logarithms of its doses and the reciprocal of the bleeding and coagulating times in mice. Obvious hemostatic action was observed after mice had been administered in ip and po respectively for 0.5h and 1h. The hemostatic time of the former was 6h and the latter 12h. The LD50 of StE in po was 2.652 +/- 0.286 g/kg, while in ip 1.945 4/- 0.207 g/kg.