• Journal of Veterinary Clinics
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• v.22 no.4
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• pp.377-381
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• 2005

We tried to extract bone morphogenetic protein (BMP) from the freeze-dried bovine cortical bone (FBCB) for bone graft, which were defatted with chloroform-methanol for 20 days, freeze-dried at $-80^{\circ}C$ for 7 days and sterilized by ethylene oxide gas. Two kg of FBCB were pulverized in a wheel mill to $0.5-2.0mm^3$ cubic in size. The bone particles were demineralized in 0.6N HCI for 10 days at chloroform-methanol$4^{\circ}C$ and defatted with chloroform-methanol for 6 hours at room temperature, which was going to be defatting and demineralized cortical bone (DDM). For extracting BMP, DDM was agitated continuously through 72 hours with magnetic stirrer at $4^{\circ}C$ into 12 times of volume of 6 M guanidine hydrochloride (Gdn-HCl) solution containing proteinase inhibitors to protect BMP such as 2mM N-ethylaleimide, 1mM iodoacetic acid, 1mM phenylmethylsulfonyl fluoride and a sterilizer, 1mM sodium azide. The extraction procedure was repeated for three times. All extracted solution was centrifuged at 10,000 rpm for 30 min and then, the supernatant was dialyzed with 12 times of volume of deionized water at $4^{\circ}C$ for 24-72 hours, which cut off below 6,000-8,000 molecular weight. The dialyzed specimen contained crude-BMP was centrifuged, freeze-dried, and weighted. Through these processing, we could obtained $84.9\%$ as FBCB, $17.8\%$ as DDM and $0.71\%$ as crude-BMP from the wet cortical bone without cancellous bone, marrow and muscles. The crude-BMP were obtained $68.3\%$ from the first extraction, $29.6\%$ from secondary and $2.1\%$ from tertiary, respectively. It was suggested that high yield of crude-BMP migth be explained by three-time repetition of the extraction processing for crude-BMP with Gdn-Hcl sol.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.18 no.2
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• pp.236-244
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• 1996

The maxillary sinus elevation for simultaneous placement of dental implants and combination grafts of autogenous bone harvested from the maxillary tuberosity and demineralized freeze dried bone and HA is relatively easy and safely done under local anesthesia in out patients clinic. This article is to introduce the sinus floor elevation method which has been performed to 5 patients in the department of Dentistry/Oral & Maxillofacial Surgery, Kangnam Sacred Heart Hospital, Hallym University, from 1993.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.28 no.4
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• pp.290-301
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• 2002

The purpose of this study was to evaluate new bone formation and healing process in rat calvarial bone defects using $BioMesh^{(R)}$. membrane and DFDB. Forty eight rats divided equally into 4 groups of 1 control group and 3 experimental groups. Standardized transosseous circular calvarial defects (8 mm in diameter) were made midparietally. In the control group, the defect was only covered with the soft tissue flap. In the experimental group 1, it was filled with DFDB only, in the experimental group 2, it was covered $BioMesh^{(R)}$. membrane only, and in the experimental group 3, it was filled DFDB and covered with membrane. At the postoperative 1, 2, 4, 8 weeks, rats were sacrificed and histologic and histomorphometric analysis were performed. These results were as follows. In histomorphometric analysis, It showed the greatest amount of new bone formation through experimental in the experimental group 3 (P<0.001). The amount of new bone formation at the central portion of the defect was greater in the experimental group 3 than experimental group 2. $BioMesh^{(R)}$. membrane began to resorb at 1 week and resorbed almost completely at 8 weeks after operation. The collapse of membrane into the defect was observed through the experimental periods in the experimental group 2. In the area of collapsed membrane, new bone formation was restricted. These results suggest that maintenance of some space for new bone to grow is required in the use of $BioMesh^{(R)}$. membrane alone in the defect. It is also thought that use of the membrane may promote new bone growth in DFDB graft.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.28 no.3
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• pp.188-195
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• 2002

The purpose of this investigation was to evaluate the relationship between the hydration time of demineralized freeze-dried bone (DFDB) and early new bone formation in rat calvarial defects filled with DFDB. Rats (n = 43) were divided into 4 experimental groups. Standard, transosseous circular defects of the calvaria were made midparietally. In experimental group 1, the defect was grafted immediately after soaking the DFDB. In experimental group 2, the defects were grafted with DFDB after soaking the DFDB for 10 minutes. In experimental groups 3 and 4, the defects were filled after soaking the DFDB for 30 and 60 minutes, respectively. Graft sites were analyzed histologically after healing periods of 1, 2, or 4 weeks. Each group showed similar bone regeneration at each time point by histological analysis. The results of this study were as follows: 1. After 1 week, a significant amount of inflammation, granulation tissue, and edema were found. A small amount of bone was seen, but the amount of bone did not differ between groups. 2. After 2 weeks, a small amount of new bone formation and DFDB resorption were observed. 3. After 4 weeks, a greater amount of new bone formation was observed. The greatest amount of bone formation occurred in experimental group 4 after 4 weeks. We conclude that the hydration time of DFDB does not affect new bone formation and that it is very important to control inflammation in bone grafting.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.17 no.4
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• pp.337-349
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• 1995

Periosteum in general is described as a specialized fibrous membrane of mesenchymal origin consisting of two basis layers : outer fibrous layer consists of irregularly arranged dense connective-tissue with fibroblasts, and inner osteogenic or cambial layer is composed of more loosely arranged fibers, greater vascularity and flatted spindle-shaped pre-osteoblasts. This periosteum may serve in controlling bone growth, especially mandibular growth has been emphasized. But, the periosteum enwrapping the facial skeleton have been studied for many years leaving a controversy in opinion regarding the function of these structures. We evaluated the bone formation activity of te periosteum in allogeneic bone grafts which bones are made of freeze-dried preparation preoperatively. We made the calvarial bone defects, 5 ${\times}$ 7mm sized, amd grafted with allogeneic bone in rats, which a half of specimens has dissected the overlying periosteum and a rest intacted. After bone grafting, we evaluated the capacity ofbone formation of periosteum, 1, 2, 4, 6, 8 weeks postoperatively. There are subtle differences of bone formation during early healing period after demineralized allogeneic bone grafting between control groups with periosteum and experimental groups without periosteum.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.17 no.3
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• pp.239-252
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• 1995

The present study was aimed to evaluate the effect of demineralized freeze-dried bone (DFDB) and resorbable hydroxyapatite (RHA) on bone formation in the extracted socket. The lower left and right 2nd and 3rd premolar were extracted in adult dogs. The one group was grafted with DFDB into the extracted socket, and the other group grafted with RHA. The extracted socket was sutured without any graft materials as control. The animals were killed on the 1st, 2nd, 4th, and 8th week after the graft for macroscopic and microscopic examination. Results obtained were as follows : 1. Macroscopically, nor infection of the graft site and dislodgement of the grafted material were noted in any animals used. 2. Young trabeculae of osteoid were formed in the socket wall in control group at 2 weeks after the graft. Osteoid tissue was formed in DFDB group at 1 week after graft, and a fine osteoid tissue was grown through the RHA particles in RHA group at 2 weeks graft. 3. The grafted groups showed more rapid bone formation than the control. Between the grafted groups, DFDB group showed more rapid formation than RHA group, DFDB group showed osteoinductive bone formation and RHA group showed osteoconductive bone formation. These results suggest that DFDB and RHA are useful to preserve the alveolar bone and to improve new bone formation by immediate grafting into the extraction sockets.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.18 no.3
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• pp.371-377
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• 1996

Bone graft has been used to repair one defect caused by disease and trauma, congenital and acquired deformities. Graft materials are autogenous bone, allogenic bone, xenogenic bone, synthetics. Autogenous bone graft is the most superior to other materials for immunologic reaction, compatibility to host tissue, and revascularization. However, autogenous bone graft is required for additional operation and the amount of taking is limited. Autografts are obtained at own expense and also limited in size, shape. In order to compensate these problems, allogenic bone graft has been used increasingly. But allogenic bone graft encounters immunologic complications. Therefore, it has been used after freezing, lyophilization, or demineralization. Allogenic bone processed by only lyophilization includes potential antigenic properties on its surface, therefore it is demineralized to deplete immunologic reaction. Demineralized bone releases BMP and helps the mesenchymal cells transform to the chondroblast to produce cartilage and bone. This reaction is called osteoinducation. Many authors have reported that mineralized lyophilized bone had less antigenicity clinically and favorable bony consideration with host bone. In our department from 1995 to now, we have used banked allogenic bone graft that has been prepared from Wonkwang Bone Bank in 5 cases and mineralized lyophilized bone graft in 2 cases to reconstruct the maxillofacial bone defect after tumor resection and cyst enucleation and cleft alveolus. We will report with literature review that the result is favorable functionally and esthetically.

• Journal of Periodontal and Implant Science
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• v.42 no.6
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• pp.224-230
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• 2012

Purpose: Various bone graft materials have been used for periodontal tissue regeneration. Demineralized freeze-dried bone allograft (DFDBA) is a widely used bone substitute. The current widespread use of DFDBA is based on its potential osteoinductive ability. Due to the lack of verifiable data, the purpose of this study was to assess the osteoinductive activity of different DFDBAs in vitro. Methods: Sarcoma osteogenic (SaOS-2) cells (human osteoblast-like cells) were exposed to 8 mg/mL and 16 mg/mL concentrations of three commercial types of DFDBA: Osseo+, AlloOss, and Cenobone. The effect of these materials on cell proliferation was determined using the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The osteoinductive ability was evaluated using alizarin red staining, and the results were confirmed by evaluating osteogenic gene expression using reverse transcription polymerase chain reaction (RT-PCR). Results: In the SaOS-2 cells, an 8 mg/mL concentration of Osseo+ and Cenobone significantly increased cell proliferation in 48 hours after exposure (P<0.001); however, in these two bone materials, the proliferation of cells was significantly decreased after 48 hours of exposure with a 16 mg/mL concentration (P<0.001). The alizarin red staining results demonstrated that the 16 mg/mL concentration of all three tested DFDBA induced complete morphologic differentiation and mineralized nodule production of the SaOS-2 cells. The RT-PCR results revealed osteopontin gene expression at a 16 mg/mL concentration of all three test groups, but not at an 8 mg/mL concentration. Conclusions: These commercial types of DFDBA are capable of decreasing proliferation and increasing osteogenic differentiation of the SaOS-2 cell line and have osteoinductive activity in vitro.

• The Journal of the Korean dental association
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• v.32 no.4 s.299
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• pp.285-297
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• 1994

• Journal of Periodontal and Implant Science
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• v.27 no.4
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• pp.845-866
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• 1997

Several experimental studies showed that the application of small amounts of electric current to bone stimulated osteogenesis at the site of the cathode and suggests that the application of electrical currents to periodontal defects could promote bone and cementum formation. The purpose of this study was to determine the effect of direct microcurrent to the periodontal regeneration of class III furcation defects in dogs. Class III furcation defects were surgically created on the third and the fourth premolars bilaterally in the mandibles of nine mongrel dogs. Experimental periodontitis were induced by placing small cotton pellets into the created defects for 3 weeks. The experimental sites were divided into three groups according to the treatment modalities: Group I-surgical debridement only; Group II-allogenic demineralized freeze dried bone grafting; Group III-allogenic demineralized freeze dried bone grafting and electrical stimulation. For fluorescence microscopic evaluation, calcein, oxytetracycline HCI and alizarin red were injected 2, 4 and 8 weeksfS days prior to sacrifice) after surgery. The animals were sacrificed in the 1st, 2nd, 4th and 8th week after periodontal surgery and the decalcified and undecalcified specimens were prepared for histological and histometrical examination. After the first and the second weeks, gingival recession was more severe in group I than groups II and III. After the fourth and the eighth weeks, there was no difference in the width of junctional epithelium and connective tissue attachment among the three groups, but the width of connective tissue attachment increased in group II at the eighth week, compared to the fourth week. The amount of bone repair in new attachment was significantly greater in group III, compared to groups I and II. New attachment formation was significantly greater in group III, compared to groups I and group II. These results suggest that electrical stimulation using microcurrent generator could be a useful tool for periodontal regenerative therapy in class III furcation defect.