• Maxillofacial Plastic and Reconstructive Surgery
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• v.31 no.3
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• pp.198-206
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• 2009

Purpose: The aims of this study were to assess the in vitro co-culturing pattern of isolated skin-derived precursor cells (SKPs) with a mixed demineralized bone (DMB) and fibrin glue scaffold and to evaluate in vivo osteogenesis after transplantation of autogenous SKPs with a these mixed scaffold in the animal's mandibular defects. Materials and Methods: We isolated SKPs from the ears of adult 4 miniature pigs. The isolated SKPs were co-cultured with a mixed DMB and fibrin glue scaffold in a non-osteogenic medium for 1, 2, and 4 weeks. Histological characteristics of in vitro co-cultured cells and scaffold were evaluated. $1{\times}10^7\;cells/100\;{\mu}l$ of autogenous porcine SKPs were grafted into the mandibular defects with a DMB and fibrin glue scaffold. In the control sites, only a scaffold was grafted, without SKPs. After two animals each were euthanized at 2 and 4 weeks after grafting, the in vivo osteogenesis was evaluated with histolomorphometric and osteocalcin immunohistochemical studies. Results: Homogeneously shaped skin-derived cells were isolated from porcine ear skin after 3 or 4 weeks of primary culture. In vitro osteogenic differentiation of SKPs was observed after co-culturing with a DMB and fibrin glue scaffold in a non-osteogenic medium. Von Kossa-positive bone minerals were also noted in the co-cultured medium at 4 weeks. As the culture time progressed, the number of observable cells increased. Trabecular new bone formation and osteocalcin expression were more pronounced in the SKP-grafted group compared to the control group. Conclusion: These findings suggest that autogenous SKP grafting with a DMB and fibrin glue scaffold can serve as a useful alternative to bone grafting technique.

• Journal of Periodontal and Implant Science
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• v.23 no.3
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• pp.577-594
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• 1993

The ultimate goal of periodontal therapy is the regeneration of the periodontium that have been destroyed as a result of periodontal disease. This study were done in order to determine the healing status of periodontium under Polytetrafluoroethylene and millipore fillter combined with fibrin and the effect of the guided tissue regeneration procedures were performed as follows : 1) flap operation using PTFE membrane(control group) 2) flap operation using PTFE membrane which was fixed with fibrin(experimental group 1) 3) flap operation using millipore filter which was fixed with suture(experimental group II) 4) flap operation using millipore filter which was fixed with fibrin(experimental group II) After 1, 2, 4, 8, 12 weeks, dogs were sacrificed by perfusion technique and tissue block was excised including the tooth and prepared for light microscope with H-E & Masson’s trichrome staining. The result were as follows : In control and experimental group, there is no siginificant difference on epithelial cell down growth within 1st week, but more epithelial cell downgrowth in millipore or millipore combined with fibrin group. In this experiment, there were no significant difference in new cementum and alveolar bone formation whether PTFE membrane was fixed with suture or fibrin. In control and each experimental group, bone maturation appeared in 4 weeks, bone width increased bucco-lingually in control and experimental 1 group especially. Both control group and experimental group showed mild mew cementum formation on root surface and irregular arrangement of collagen fiber at 4 weeks, that showed obvious increased cementum formation at 8 weeks, and that was observed the functional arrangement of collagen fiber between new cementum and new alveolar bone at 12 weeks.

• Journal of dental hygiene science
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• v.13 no.2
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• pp.158-164
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• 2013

The most frequently encountered problems at fixture-implantation sites are lack of adequate bone and proximity to anatomic structures. It is generally accepted that growth factors play an essential role in the healing process and tissue formation, and they have become the focus of grafting materials research. The granules in platelets contain high concentrations of various growth factors. In particular, platelet-rich fibrin (PRF) is a second-generation platelet concentrate that allows the production of fibrin membranes enriched with platelets and growth factors from an anticoagulant-free blood harvest. This study investigated the in vitro effects of PRF on osteoblasts, in terms of the key cellular functions, and especially the effects on two growth factors, the homodimer of platelet-derived growth factor subunit B (BPDGF-BB) and transforming growth factor (TGF)-${\beta}1$, which are associated with wound healing and regeneration (i.e., proliferation and differentiation). The following parameters were investigated: PDGF-BB and TGF-${\beta}1$ levels in PRF, cell viability, alkaline phosphatase (ALP) activity, type 1 collagen synthesis, and the expressions of osteoblast differentiation markers (ALP and runt-related transcription factor 2) and bone matrix proteins (type 1 collagen). The release of autologous growth factors from PRF was maintained for a reasonable period of time, and exerted positive effects on the proliferation and differentiation of osteoblasts. The use of PRF thus appears to be a promising method for enhancing bone healing and remodeling.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.15 no.1
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• pp.49-61
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• 1993

The transplantation of cartilage, especially auricular cartilage, has assumed a role of importance in the field of plastic and reconstructive surgery. From long years ago, many reports have appeared in the literature describing the experimental and clinical results of the use of cartilage. At present, the evidence for survival of autograft of cartilage is admitted, But, the results for interrelationship between the bone and cartilage grafts with or without perichondrium is not so conclusive. The purpose of this study were observed as to whether autogenous cartilage grafts were fixed by means of tie with 4-0 vicryl and fibrin adhesive on the femur or microscopic findings of union state in 16 rabbits. We sacrified the experimental animals after 1, 2, 4, 6 weeks postoperatively and made the specimens as a routine laboratory procedures and stained with Hematoxylin-Eosin stain, Verhoeff-van Gieson elastic fiber stain, and alcian blue periodic acid-Schiff(AB-PAS) for mucopolysaccharide. Histologic evaluation was performed under microscope. The obtaind results were as follows : 1. Fibrous union was formed between the grafting cartilage and the femur, nor any findings of calcification and formation of new bone. 2. Partial fibrous adhesion was observed in fibrin adhesive groups on 6 weeks postoperatively. 3. Appositional growth has performed more in fibrin adhesive groups than tie groups. 4. There are little difference in both for new copillary proliferation and fibroblast activations. 5. Degenerative changes have apperared more in tie groups than adhesive groups, but not related to the healing periods.

• Korean Journal of Food Science and Technology
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• v.29 no.4
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• pp.817-822
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• 1997

Inhibitory mechanism of the anticoagulant polysaccharide purified from the fruit body of Coriolus versicolor was investigated in this paper. The anticoagulant polysaccharide (CV-40-Va-1) was proposed to have functions of the inhibition of intrinsic pathway in the blood coagulation pathway together with the interuption of a human platelet aggregation induced by von Willebrand factor (vWF). CV-40-Va-I inhibited other factors of the coagulation cascade such as factor VIII, IX, and as well as thrombin. Especially, CV-40-Va-I inhibited the fibrin formation mediated by thromin, however the polysaccharide did not affect the fibrin formation directly but affected the anticoagulant activity through the activation of antithrombin III. The sulfation of the anticoagulant polysaccharide increased the anticoagulant activity, showing that the sulfate concentration of anticoagulant polysaccharide was important factor in the blood coagulation cascade. Low molecular weight subfraction (MW 1,000) obtained by partial hydrolysis of the CV-40-Va-1 generated potent antiplatelet activity, but showed decreased anticoagulant activity.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.4
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• pp.340-349
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• 2007

Free flap transplantation with microvascular anastomosis has been successfully performed by development of surgical technique, materials and postoperative monitoring equipments of flap. But success rate of microvascular anastomosis is influenced by various factors, and failure rate is about 5-10%. The most influential factor for success rate is surgical technique and other factors that influence failure of microvascular anastomosis are ischemic time of free flap, thrombus formation of anastomosis region and vascular spasm. In this study, vascular patency and thrombus formation in experimental micro-venous anastomosis, and endothelial repair were observed with histologic analysis, scanning electron microscopy, transmission electron microscopic examination. The results were obtained as follows: 1. In vascular patency test in 30 minute and 7 days after micro-venous anastomosis with heparin irrigation, all of 12 anastomosis site were good vascular patency. 2. In thrombus formation in 2 weeks group(Experimental I), 2 site of 6 cases were observed thrombus, and in 4 weeks group(Experimental II), 1 site of 6 cases were observed thrombus. 3. In histologic examination, normal vein(Control Group) showed continued internal elastic lamina, well formed thick smooth muscle layer and connective tissue. The group of 2 weeks after microvenous anastomosis(Experimental I) showd locally recovered internal lamina, discontinued internal lamina, disorganized smooth muscle cells and granulation tissue around suture silk. In the group of 4 weeks after micro-venous anastomosis(Experimental II), anastomosis site showed almostly continued internal lamina, disorganized smooth muscle cells and cicartrized tissue around suture silk. 4. In scanning electron microscope examination in 2 weeks(Experimental I) after micro-venous anastomosis, mesh fibrin formation showed near to endothelial cells, and in 4 weeks after micro-venous anastomosis(EXperimental II), numerous blood cells and fibrin mesh formation was seen associated with irregular endothelial cell arrangement. 5. In transmission electron microscope examination in 2 weeks after micro-venous anastomosis(Experimental I), irregular arrangement of smooth muscle cells was seen adjacent to collagenized tissue around suture silk. In 4 weeks after micro-venous anastomosis(Experimental II), denuded venous wall composed of relatively well arranged smooth muscle cells was covered by endothelial cells, but fibroblast cells and foreign body giant cells near to suture silk was remained. From the results obtained in this study, results of good vascular patiency and anti-thrombotic effect of heparin were obtained as a local irrigation solution, and repair of venous endothelial cell was observed in 2 weeks after micro-venous anastomosis.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.34 no.6
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• pp.391-397
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• 2012

Purpose: The purpose of this study was to evaluate the effectiveness of the platelet-rich fibrin (PRF) used in combination with the porcine cancellous bone as a scaffold, in promoting bone regeneration in the bone defects ofthe rabbit calvaria. Methods: Ten rabbits were used in the study. Three round-shaped defects (diameter 8.0 mm) were created in the rabbit calvaria and were filled with nothing (control group), porcine cancellousbone (Experimental Group 1, porcine bone) and PRF-mixed porcine cancellous bone (Experimental Group 2). TS-GBB is a xenogenic bone-substitute product comprised of a high heat-treated mineralized porcine cancellous bone. Animals were sacrificed at 6 weeks and 12 weeks for the histological and radiographic evaluations. Results: In the micro computed tomography and histological results, the experimental groups 1 and 2 showed more bone formation, remodeling, and calcification than the control group. The new bone formation ratio showed theGroup 2 to be larger than Group 1 at6 and 12 weeks. However, there was no significant difference between the experimental groups 1 and 2 in the new bone formation area, at the 6 and 12 weeks (P>0.05). Conclusion: The PRF-mixed group showed more bone formation than the porcine cancellousbonegroup (TS-GBB), butthere was a no significant difference. The PRF may not lead to enhanced bone healing when grafted with the porcine cancellous bone.

• Journal of Applied Biological Chemistry
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• v.65 no.4
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• pp.239-245
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• 2022

Although normal activation of platelets is important in the process of hemostasis, excessive or abnormal activation of platelets can lead to cardiovascular diseases. Therefore, the discovery of novel substances capable of regulating or inhibiting platelet activation may be helpful in the prevention and treatment of cardiovascular diseases. Artemether is a derivative of artemisinin, known as an active ingredient of Artemisia annua, which has been reported to be effective in treating malaria, and is known to function through antioxidant and metabolic enzyme inhibition. However, the role of artemether in platelet activation and aggregation and the mechanism of action of artemether in collagen-induced human platelets are not known until now. This study investigated the effects of artemether on platelet activation and thrombus formation induced by collagen. As a result, cAMP level was significantly increased by artemether, and VASP and IP3R, substrates of cAMP-dependent kinase, were phosphorylated. IP3R phosphorylation by Artemether inhibited Ca²⁺ recruitment into the cytoplasm, and phosphorylated VASP inhibited fibrinogen binding by inactivating αIIb/β3 located on the platelet membrane. Consequently, artemether inhibited thrombin-induced fibrin clot formation. Therefore, we propose that artemether can act as an effective prophylactic and therapeutic agent for cardiovascular diseases caused by excessive platelet activation and thrombus formation.

• Journal of Applied Biological Chemistry
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• v.63 no.3
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• pp.235-241
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• 2020

An essential component of the hemostatic process during vascular damage is platelet activation. However, many cardiovascular diseases, such as atherosclerosis, thrombosis, and myocardial infarction, can develop due to excessive platelet activation. Isoscopoletin, found primarily in plant roots of the genus Artemisia or Scopolia, has been studied to demonstrate potential pharmacological effects on Alzheimer's disease and anticancer, but its mechanisms and role in relation to thrombus formation and platelet aggregation have not yet been discovered. This research investigated the effect of isoscopoletin on collagen-induced human platelet activation. As a result, isoscopoletin strongly increased cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) levels in a concentration-dependent manner. In addition, isoscopoletin greatly phosphorylated inositol 1,4,5-triphosphate receptor (IP₃R) and vasodilator-stimulated phosphoprotein (VASP), known substrates of cAMP-dependent kinase and cGMP dependent kinase. Phosphorylation of IP₃R by isoscopoletin induced Ca²⁺ inhibition from the dense tubular system Ca²⁺ channels, and VASP phosphorylation was involved in fibrinogen binding inhibition by inactivating αIIb/β₃ in the platelet membrane. Isoscopoletin finally reduced thrombin-induced fibrin clot production and finally reduced thrombus formation. Therefore, this research suggests that isoscopoletin has strong antiplatelet effects and is likely to be helpful for thrombotic diseases involving platelets by acting as a prophylactic and therapeutic agent.

• BMB Reports
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• v.29 no.6
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• pp.500-506
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• 1996

We have isolated a water-extracted novel regulator for blood coagulation from an earthworm, Lumbricus rubellus. As a folk remedy, the earthworm has been known to facilitate blood circulation. After complete heat inactivation of endogenous proteases in the earthworm, an anticoagulant(s) was purified through ammonium sulfate fractionation and three consecutive gel permeation chromatography of Sephacryl S-300, Sephadex G-75, and G-150 by measuring activated partial thromboplastin time (APTT) The anticoagulant was further purified to 2,800 fold with a C4 reversed-phase HPLC This activity was stable under heat ($100^{\circ}C$ for 30 min) and acidic conditions (0.4 N HCl). The effects of this partially purified anticoagulant on thrombin were observed with various substrates such as N${\alpha}$-benzoyl-DL-arginine-p-nitroanilide (BApNA), H-D-phenylalanyl-L-pipecoyl-L-arginine-p-nitroanilide (S-2238), N${\alpha}$-p-tosyl-L-arginine methyl ester (TAME), and fibrinogen as a natural substrate. Only TAME hydrolysis, due to an esterase activity of the enzyme, was inhibited among the chromogenic substrates. In addition, the anticoagulant not only inhibited the conversion of fibrinogen to fibrin but also prolonged the fibrin clot formation monitored with the in vitro coagulation test. Based on these observations, we suggest the significance of measuring the ability of antithrombotic drugs to inhibit the esterase activity of thrombin. In this report, it was also shown that the earthworm indeed contained a water-extractable, heat- and acid-stable anticoagulant which could be used as a novel antithrombotic agent.