• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.26 no.1
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• pp.24-39
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• 2000

Various methods and graft materials have been used to fill in the defect adjacent to the implants and considered as clinically acceptable. But it is not clear whether the regenerated bone increases the implant-bone contact and supports the implant. The purpose of this study is to evaluate regenerated bone surrounding implants using bone morphogenetic protein(BMP) and demineralized freeze-dried bone(DFDB), and the interfaces between implants and regenerated bone. bBMP was extracted and partially purified from the bovine bone matrix using heparine chromatography. Demineralized freeze-dried bone was made from the dog. Inactive insoluble collagenous bone matrix(IBM) of dog was used as carrier of bBMP. Interfaces of titanium coated epoxy resin implants were processed for demineralized section for transmission electron microscopy(TEM) and those of screw type implants were for nondemineralized section for light and fluoromicroscopic examination. Implants were inserted in the inferior border of mandible of adult dogs and artificial bony defects($3{\times}3{\times}4mm$) were made at the mesial and distal side of implants. Defects were filled with BMP(BMP group) and DFDB(DFDB group). For the fluoromicroscopic examination, the fluorescent dyes(oxytetracycline, calcein green, alizarin red) were injected 2, 4, 6, 8, 12 weeks after implantation. The experimental animals were sacrificed at the 6th and the 12th week and their mandible were extirpated and processed for examination with light microscopy, fluoromicroscopy and TEM. The obtained results were as follows : 1. By the light microscopic findings, the defects were filled with woven bone at the 6th week and compact bone at the 12th week, and the osseointegrations were seen in both groups. There was no histological difference between them. 2. On the basis of the histomorphometric analysis, BMP group(6th week: 40.25%, 12th week: 56.04%) had higher bony contact ratio than DFDB group(38.37%, 42.63%). There was significant difference between two groups at the 12th week(p<0.05). 3. The amount of bone formation in BMP group was more prominent than in DFDB group. Significant difference was noted among two groups at the 6th and the 8th week(p<0.05). 4. By the transmission electron microscopic findings, $0.4-2{\mu}m$ soft tissue layer was found in adjacent to the interfaces and over the collagen fibrils of bone at the 6th week. However, about 100nm amorphous layer was noted at the interface or collagen fibrils directly extended to the titanium surface at the 12th week. There was no significant difference between two groups. 5. These results suggest that BMP and DFDB can be used as good graft materials in the regeneration of bone adjacent to implant, and BMP is more valuable as a bone inducer than DFDB.

• Journal of Periodontal and Implant Science
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• v.31 no.1
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• pp.57-74
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• 2001

The purpose of this study was to evaluate the adjunctive combined effect of demineralized freeze-dried bone allograft(DFDB) in guided bone regeneration on supra-alveo-lar peri-implant defect. Supra-alveolar perio-implant defects, 3mm in height, each including 4 IMZ titanium plasma-sprayed implants were surgically created in two mongrel dogs. Subsequently, the defects were treated with 1 of the following 3 modalities: Control) no membrane or graft application, Group1) DFDB application, Group2) guided bone regeneration using an expanded polytetra-fluoroethylene membrane, Group3) guided bone regeneration using membrane and DFDB. After a healing period of 12-week, the animals were sacrificed, tissue blocks were harvested and prepared for histological analysis. Histologic examination were as follows; 1. New bon formation was minimal in control and Group 1, but considerable new bone formation was observed in Group 2 and Group 3. 2. There was no osteointegration at the implant-bone interface in the high-polished area of group2 and Group 3. 3. In fluorescent microscopic examination, remodeling of new bone was most active during week 4 and week 8. There was no significant difference in remodeling rate between group 2 and group 3. 4. DFDB particles were observed, invested in a connective tissue matrix. Osteoblast activity in the area was minimal. The results suggest that guided bone regeneration shows promising results in supra-alveolar peri-implant defects during the 12 week healing period although it has a limited potential in promoting alveolar bone regeneration in the high-polished area. There seems to be no significant adjunctive effect when DFDB is combined with GBR.

• 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.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.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.341-356
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• 1995

The purpose of this study was to evaluate the effect of demineralized freeze dried bone and demineralized bone gel with guided tissue regeneration treatment around titanium implants with dehisced bony defects and also evaluate space maintaining capacity of demineralized bone gel type and DFDB powder type under e-PTFE membrane. In 3 Beagle dogs, mandibular premolar was extracted and four peri-implant osteotomies were formed for dehiscence. After insertion of implants, the four peri-implant defects were treated as follows. 1) In control group. no graft material and barrier membrane were applied. 2) In experimental group.1, the site was covered only with the e-PTFE membrane. 3) In experimental group 2,received DFDB powder and covered by the e-PTFE membrane. 4) In experimental group 3, demineralized bone gel and e-PTFE membrane were used. By random selection, animals were sacrificed at 4, 8, 12 weeks. The block sectioned specimens were prepared for decalcified histologic evaluation(hematoxylin and eosin staining) and undecalcified histologic evahiation(Von Kossa's and toluidine blue staining) with light microscopy. The results of this study were as follows. 1) In control group, there was a little new bone formation and connective tissue was completely filled in the defect area. 2) Experimental group 1 showed lesser quantity of bone formation as compared to the bone grafted group. Thin vertical growth of new bone formation around implant fixture was shown. 3) Experimental group 2 showed thick bucco-lingual growth of new bone formation and grafted bone particles were almost resorbed in 12 week group. 4) In experimental group 3, most grafted bone particles were not resorbed in 12 week group and thick bucco-lingual bone formation was shown in dehisced defect base area. 5) There was no remarkable differences in space making capacity and new bone formation procedure between demineralized freeze-dried bone powder type and demineralized bone gel type.

• 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 Periodontal and Implant Science
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• v.36 no.1
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• pp.223-237
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• 2006

The purpose of the present study was to evaluate the effect of bone graft materials including deproteinized bovine bone(DBB), demineralized freeze-dried bone(DFDB), freeze-dried bone(FDB) on bone formation in guided bone regeneration using perforated titanium membrane(TM). 16 adult male rabbits(mean BW 2kg) were used in this study and 4 rabbits allotted to each test group. Intramarrow penetration(diameter 6.5mm) was done with round carbide bur on calvaria to promote blood supply and clot formation in the wound area. The test groups were devided into 4 groups as follows: TM only(test 1), TM +DBB(test 2), TM +DFDB(test 3), TM +FDB(test 4). Perforated titanium membrane was contoured in rectangular parallelepiped shape(0.5mm pore diameter, 10mm in one side, 2mm in inner height), filled the each graft material and placed on the decorticated carvaria. Perforated titanium membrane was fixed with resorbable suture materials. The animals were sacrificed at 2, 8 weeks after the surgery. Non-decalcified preparations were routinely processed for histologic analysis. The results of this study were as follows: 1. Perforated titanium membrane was biocompatible. 2. Perforated titanium membrane had capability of maintaining the space during the healing period but invasion of soft tissue through the perforations of titanium membrane decreased the space available for bone formation. 3. In test 1 group without bone graft material, the amount of bone formation and bone maturation was better than other test groups. 4. Among the graft materials, the effect of freeze-dried bone on bone formation was best. 5. In the test groups using deproteinized bovine bone, demineralized freeze-dried bone, bone formation was a little. The spacemaking capability of the membrane may be crucial for bone formation. The combined treatment with the perforated titanium membrane and deproteinized bovine bone or demineralized freeze-dried bone failed to demonstrate any added effect in the bone formation. Minimization of size and numbers of perforations of titanium membrane or use of occlusive titanium membrane might be effective to acquire predictable results in the vertical bone formation.

• 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 academy of Pediatric Dentistry
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• v.27 no.2
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• pp.318-332
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• 2000

The purpose of this study was to investigate the effects of demineralized freeze-dried bone (DFDB) on mechanically exposed pulp of dog by evaluating the pulpal inflammation and healing process, formation of dental hard tissue, and structural changes of fibroblasts of the remaining pulp tissue. Teeth of 4 dogs, weighing 10kg, were used in this study. Class V cavities were prepared followed by exposed the pulp tissue mechanically by sterilized round bur. In control group, exposed pulps were capped with calcium hydroxide paste followed by sealed with IRM. In experimental groups, the exposed pulps of one group were capped with the collagen and those of the other group were capped with DFDB. All cavities were sealed with same manor as control group. The animals were sacrificed at the intervals of 3, 7, 14, and 28 days for histopathlogic evaluation. The specimens were observed by the light microscope and trans-electron microscope. The results were as follows: 1. Pulp necrosis was not observed in all groups. Inflammatory response was disappeared from 1 week in control group and group 2. But it was not disappeared until 2 weeks and also irregular arrangement of odontoblasts was showed at the lateral walls of root canal just beneath the amputated site of the pulp in group 1. 2. Dentinal bridge was formed incompletely at 2 weeks but it was formed completely at 4 weeks in control group. Odontoid tissue was also found in control group at 4 weeks from treatment. Amputated site of pulp was encapsulated with fibrous tissue and odontoblast and dentinal bridge was not found in group 1. Preodontoid tissue and reparative dentin which were formed by odontoblast differentiated around DFDB were found, but dentinal bridge was not found in group 2. 3. Cell with large basophillic-stained nuclei infiltrated to amputated site and DFDB at 1 week from treatment in control group and group 2. They were found more in group 2 than in control group. Odontoblasts arranged more regularly and reparative dentin was found more as time elapsed. 4. Dentin-formative odontoblasts which showed ultramicrostructure of cytoplasm with polarized nucleus, rEM, Golgi complex, secretory granules, secretion of organic matrix in control of group and group 2. In regards to above results, the demineralized freeze-dried bone(DFDB) induce odontoblastic differentiation and further come up to the dentin formation in amputated pulp.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.26 no.6
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• pp.557-564
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• 2000

The bone graft materials can be grossly divided into autogenous bone, allogenic bone, xenogenic bone, and alloplastic material. Much care was given to other bone graft materials away from autogenous bone due to its additional operation for harvesting, delayed resorption and limitation of quantity. Demineralized freeze-dried bone(DFDB) and hydroxyapatite are the representatives of bone graft materials. As resorbable hydroxyapatite is developed in these days, the disadvantage of nonresorbability can be overcome. So we planned to study on the strength and the bone formation at the rats calvarial defects of DFDB graft and those of the composite graft with DFDB and resorbable hydroxyapatite. We used the 16 male rats weighting range from 250 to 300 gram bred under the same environment during same period. After we made the 6mm diameter calvarial defect, we filled the DFDB in 8 rats and DFDB and resorbable hydroxyapatite in another 8 rats. We sacrificed them at the postoperative 1 month and 2 months with the periostium observed. As soon as the specimens were delivered, we measured the compressive forces to break the normal calvarial area and the newly formed bone in calvarial defect area using Instron(Model Autograph $S-2000^{(R)}$, Shimadzu, Japan). The rest of the specimens were stained with H&E(Hematoxylin & Eosin) and evaluated with the light microscope. So we got the following results. 1. In every rats, there was no significant difference between the measured forces of normal bone area and those of the bone graft area. 2. In 1 month, the measured forces at DFDB graft group were higher than those of the DFDB and resorbable hydroxyapatite composite graft group(P<0.05). 3. In 2 months, there was no significant differences between the measured forces of DFDB graft group and those of the DFDB and resorbable hydroxyapatite composite graft group. 4. In lightmicroscopic examination, most of the grafted DFDB were transformed into bone in 1 month and a large numbers of hydroxyapatite crystal were observed in DFDB and resorbable hydroxyapatite composite graft group in 1 month. 5. Both group showed no inflammatory reaction in 1 month. And hydroxyapatite crystals had a tight junction without soft tissue invagination when consolidated with newly formed bone. 6. In both groups, newly formed bone showed the partial bone remodeling and the lamellar bone structures and some of reversal lines were observed in 2 months. From the above results, it is suggested that DFDB and resorbable hydroxyapatite composite graft group had a better resistance to compressive force in early stage than DFDB graft group, but there would be no significant difference between two groups after some period. And it is suggested that the early stage of bone formation procedure of DFDB and resorbable hydroxyapatite composite graft group was slight slower than that of DFDB graft group, but there would be no significant difference between two groups after some period.