• Journal of Veterinary Clinics
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• v.31 no.3
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• pp.170-174
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• 2014

This study evaluated the efficacy of soluble canine small intestinal submucosa gel in comparison to bovine thrombin in activating rabbit platelet rich plasma (PRP) by detecting growth factors. PRP from rabbits was activated by using soluble canine SIS gel, bovine thrombin, or both. The surface morphology of each group of samples was examined by scanning electron microscopy. The release of transforming growth factor (TGF)-${\beta}1$ from each set of samples was measured over 7 days using enzyme-linked immunosorbent assay. The PRP-canine SIS gel group exhibited the highest total amount of released TGF-${\beta}1$. However, there were no significant differences between any groups. The use of soluble type of canine SIS gel could be an effective alternative to bovine thrombin.

• Journal of Biomedical Engineering Research
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• v.16 no.3
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• pp.367-375
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• 1995

Ampicillin sodium (AMP-Na) was loaded Into fibrin glue (FG) in two different ways and was tried to achieve sustained release from FG. One was loading of AMP-Na in a simple mixing and the other was loading of bovine serum albumin (BSA) microspheres which contained ANP-Na. In case of simple mixing, the release control of AWP-Na from FG was tried by variation of FBNG concentration, but failed. However, the loading of BSA mlcrosphere containing ANP-Na into FG showed sustained re- lease of AMP-Na, especially when microsphere was crosslinked with glutaraldehyde (tO.9 : 33hr). The maximum adhesive strength of FG showed at concentration of FBWG and thrombin, 5.0 % and 25-50 NIHU/ml, respectively. The concentration of Factor Xlll (0-500 U/1g of FBNG) did not affect the adhesive strength of FG. The optimal incubation time was 60 min. The AMP-Na or BSA microsphere which was loaded into FG had no significant effect on the adhesive strength of FG.

• Tuberculosis and Respiratory Diseases
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• v.44 no.5
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• pp.1114-1124
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• 1997

Background : Endothelin(ET) is a very potent vasoconstrictive peptide produced by endothelial cells of pulmonary artery. The endothelin level was increased in plasma of primary pulmonary hypertension and acute pulmonary thromboembolism and it was suggested that the endothelin might do a critical role in the cardiopulmonary dysfunction in these two conditions. But the exact mechanism of increase of ET has not been known. In these two conditions, platelet activation and thrombosis are the main pathophysiologic findings. So there is a possibility that the platelet might stimulate endothelin secretion from endothelial cells. Therefore, we performed this study to evaluate the role of platelet and its mediators on endothelin production in bovine pulmonary artery endothelial(BPAE) cells. Method : Bovine pulmonary artery endothelial cells, ATCC certified cell line 209, were cultured and treated with human platelets($10^6{\sim}10^8/ml$), thrombin (0.1~10u/ml), TGF-${\beta}1$(1~100uM), serotonin(1~100uM), and endotoxin(1ug/ml) in a final volume of 500ul for 18 hours. Levels of ir(immunoreactive)-ET in each conditioned medium were measured by a radioimmunoassay specific for ET. Result : The increase of ir-ET levels was platelet number and time dependent over 18 hours. When washed human platelets were added($10^8/ml$), the ir-ET levels were significantly higher than that of control(p<0.05) at 8 and 18 hours after culture. Subthreshold concentration of platelets($10^7/ml$) coincubated with endotoxin(1ug/ml) or subthreshold dose of thrombin(0.1u/ml) stimulated ir-ET secretion from BPAE cells significantly(p<0.05) compared with control. Thrombin(1ug/ml, 10ug/ml) and TGF-${\beta}1$(100pM, 1000pM) significantly increased ir-ET secretion from BP AE cells(p<0.05) compared with control, but serotoin(1~100uM) and endotoxin(1ug/ml) did not stimulate the ir-ET secretion. Conclusions : Platelets stimulate endothelin secretion from bovine pulmonary artery endothelial cells. The mechanism of increase of endothelin secretion seems to be a stimulation by platelet itself or by mediators, such as TGF-${\beta}1$, secreted from activated platelets. And, in this study, the priming effect of platelets on endothelin secretion from BPAE cells could be another possibility.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.1
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• pp.44-47
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• 2000

Cell growth and DNA synthesis were studied from a cultured early- and late- pas- sage mouse aorta smooth muscle cell (MASMC) because the proliferation of vascular smooth muscle cell (VSMC) is a key factor in development of atherosclerosis. In this study, the cells were cultured in fetal bovine serum (FBS) and stimulated by growth factors such as thrombin and platelet-derived growth factor-BB (PDGF-BB). Compared to the number of early-passage MASMC (passage 3 to 9) the number of late-passage MASMC (passage 30 to 40) in a normal serum state was increased 2 fold at Day 1, 3 and 6 in culture, respectively. Incorporation of $[^3H]$ thymidine into DNA induced by serum, PDGF and thrombin in late-passage MASMC was greater than those in early-passage MASMC. We also examined whether intracellular zinc levels would be an aging factor or not. The intracellular zinc level in early- and late-passage MASMC was monitored by using the zinc probe dye N-(6-methoxy-8-quinolyl)-p-toluenesulfonamide. It is interested that late-passage MASMC increased the intracellular fluorescence level of zinc, more than the early passage MASMC did. The alterations of intracellular zinc level occur concurrently with changes in MASMC proliferation rate during aging. This data suggest that the age-associated changes in zinc concentrations may provide a new in vitro model for the study of smooth muscle cell differentiation.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.21 no.1
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• pp.81-88
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• 1999

A fibrin adhesive have been widely used in oral and maxillofacial surgery for microvascular anastomosis, autogenous chip bone grafts, many kinds of soft tissue surgery (vestibuloplasty, bleeding control after extraction, primary healing by covering of suture of a gum after the extirpation of large cysts). There are two principal components in adhesive systems biologically: lyophilized human fibrinogen and bovine thrombin. The fibrinogen component contains coagulation factor XIII and enhance the initial wound healing, which polymerizes soluble fibrin monomers into an insoluble clot. The thrombin is dissolved in a solution of calcium chloride to provide the second component. We applied fibrin adhesive, Beriplast (Behring, Behringwerke AG, D-3350, Marburg, FRD), to 4 patients for fixation of free skin grafting donors who had facial scar around eye, nose, mouth corner which received from accidents, or burn. We have experienced initial accelerated graft fixation between donor and recipient sites with no additional fixation. And It's made easy bleeding control and easy manipulation during operation. But two cases showed partial hypertrophic scar engrowth in above 3 months follow up, but no significant. Histopathological reviews in general were showed similar scar findings such as abundant collagen bundles in H&E, M/T stain, but slight positive signs in elastic and collagen antibody immunopathologic findings in hypertrophic scar cases.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.8
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• pp.1062-1068
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• 2004

Leptin has a major role in the regulation of food intake and energy homeostasis. In addition, leptin participates in many physiological functions including regulation of lipid metabolism. Bovine recombinant leptin protein was produced in E. coli cells in order to understand function of leptin in the regulation of lipid metabolism. The leptin expression vector was constructed in pGEX-4T-3 vector and transformed into E. coli BL21 cells. Expression of the GST-leptin fusion protein was induced with IPTG. The fusion protein was purified using glutathione sepharose 4B batch method, and the recombinant leptin was eluted after thrombin protease digestion. The effect of leptin on glucose transport was examined in the differentiated adipocytes of 3T3-L1 cells. Leptin had no effect on basal and insulin-stimulated glucose transport in 3T3-L1 cells (p>0.05). Effect of recombinant leptin on glucose and acetate transport was examined in adipose tissues of Korean cattle (Hanwoo). Insulin stimulated glucose transport in both intramuscular and subcutaneous adipose tissues (p<0.05), but leptin did not affect glucose transport in both adipose tissues (p>0.05). Insulin stimulated acetate transport in bovine adipose tissues (p<0.05), but leptin did not affect acetate transport (p>0.05). Northern and RT-PCR analyses showed that mRNA levels of uncoupling protein-2 were increased by leptin treatment in 3T3-L1 cells without statistical difference (p>0.05). In conclusion, bovine recombinant leptin did not affect glucose and acetate transport in both 3T3-L1 adipocytes and bovine adipose tissues, while it stimulates UCP-2 mRNA expression in 3T3-L1 cells.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.13 no.2
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• pp.129-136
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• 1991

The fibrin sealant was first designed as an alternative to surgical suture for the purpose of surface-to-surface union especially in parenchymal organs like the liver, spleen and kidney. The clinical application of currently used fibrin sealant was first introduced in 1972. The fibrin sealant consists of principal two components; lyophilized human fibrinogen and bovine thrombin. The fibrinogen component also contains coagulation factor XIII. A solution of aprotinin, an inhibitor of fibrinolysis is used to dissolve the fibrinogen and to provide the first component, and a solution of calcium chloride is also used to provide the second component. From July to December in 1990, during 6 months, we used fibrin sealant in the 28 patients of 33 various cases, in the following ways; supportive application of fibrin sealant after free autogenouse nerve graft for the repair of inferior alveolar nerve, facial nerve or accessory nerve, treament of hemangioma or lymphangioma to thrombosize and lead to the tumor shrinking, skin grafting to stimulate the adhesion and tissue repair, bone grafting in the patients of cleft alveolus, mandibular reconstruction or orthognathic surgery to facilitate the knitting of bone chips, tissue adhesion after tumor resection, radical neck dissection or flap reconstructions, and supportive adhesion of external auditory cannal after TMJ surgery via postauricular approach. No adverse effects were observed, none of the patients developed hepatitis or other blood transmitted disease, and the wound healing were acceptable.

• Archives of Plastic Surgery
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• v.39 no.6
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• pp.585-592
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• 2012

During the past decade, many studies using platelet-rich plasma (PRP) or adipose-derived stem cells (ASCs) have been conducted in various medical fields, from cardiovascular research to applications for corneal diseases. Nonetheless, there are several limitations of practical applications of PRP and ASCs. Most reports of PRP are anecdotal and few include controls to determine the specific role of PRP. There is little consensus regarding PRP production and characterization. Some have reported the development of an antibody to bovine thrombin, which was the initiator of platelet activation. In the case of ASCs, good manufacturing practices are needed for the production of clinical-grade human stem cells, and in vitro expansion of ASCs requires approval of the Korea Food and Drug Administration, such that considerable expense and time are required. Additionally, some have reported that ASCs could have a potential risk of transformation to malignant cells. Therefore, the authors tried to investigate the latest research on the efficacy and safety of PRP and ASCs and report on the current state and regulation of these stem cell-based therapies.

• Korean Journal of Veterinary Research
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• v.45 no.2
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• pp.273-278
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• 2005

This study was investigated of the bone healing capacity by autologous fibrin glue in experimental bone defect dogs. The autologous fibrin glue manufactured just before the experiment was mixed with the concentrated fibrinogen from whole blood of the experimental dog and bovine thrombin. The experimental group was constituted with seven dogs. The experimental osteotomy was performed 5 mm length in bilateral region of proximal diaphyseal fibulae. The defected regions of experimental group were filled with the autologous fibrin glue by duploject. The experimental regions had been radiographed biweekly for 16 weeks to observe new bone formation and union. Bone alkaline phophatase (BALP) in all groups was evaluated biweekly till the end of the experiment to determine osteoblast activities. New bone formation had been observed in five regions of three dogs at four weeks after the experimental treatment and in two regions of one dog at ten weeks. The other seven regions of the experimental group and control group were not observed new bone formation until the end of the experiment. BALP value in four dogs observed new bone formation was increased to 97.10 IU/L (453.96%) at two weeks after the experimental treatment. The results of this experiment were suggested that the autologous fibrin glue was moderately effective in new bone formation in dogs.

• Microbiology and Biotechnology Letters
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• v.51 no.2
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• pp.167-173
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• 2023

Proteolytic enzymes secreted by Aspergillus, as pathogenicity factors, affect blood coagulation and fibrinolysis, and therefore the target proteins of their action in the bloodstream are of significant interest. In the present study, the action of the isolated protease of A. terreus 2 on different human plasma proteins was shown. The protease of A. terreus 2 exhibited the highest proteolytic activity against hemoglobin, which was 2.5 times higher than the albuminolytic activity shown in both of the protein substrates used. In addition, the protease has significant ability to hydrolyze both fibrin and fibrinogen. However, the inability of the A. terreus 2 protease to coagulate rabbit blood plasma and coagulate human and bovine fibrinogen indicates the severity of the enzyme's action on human blood coagulation factors. It should be considered as a potential indicator of this isolated protease's participation in fungal pathogenesis. The protease shows no hemolytic activity. Furthermore, its activity is insignificantly inhibited by thrombin inhibitors, and is not inhibited by plasmin inhibitors.