• 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.

• 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.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.357-371
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• 1995

The purpose of this study was to observe the effect of $TGF-{\beta}1$ on the regeneration of bone in guided bone regeneration. Four adult dogs aged 12 to 24 months were used in this study. Experimental bone defects were created surgically with surgical bur and chisel on the 3th. premolars. In experimental group, bone defect were grafted with DFDB and $TGF-{\beta}1$. In control groups, bone defects were grafted with only DFDB. At 1,2,3 and 4 weeks, dogs were serially sacrificed and specimens were prepared with Hematoxylin-Eosin stain and Goldner's stain for light microsopic evaluation. The results of this study were as follows: 1. The infiltration of inflammatory cells was prominent in control groups at 1, 2 and 3 weeks. 2. The lining of osteoblast was observed at 2 weeks in control group, but at 1 week in experimental group. 3. In both groups, osteoid was formed at 2 weeks. In control groups, osteoid was fromed on only bone surface. but in experimental groups, osteoid were formed on both bone & DFDB surfaces. 4. In only experimental groups, The fusion of new bone & DFDB was only observed at 3 weeks. and the fusion of new bone & DFDG was more prominent at 4 weeks. But in control groups, No fusion of new bone& DFDB was oberved at 3 and 4weeks. From the above result, the $TGF-{\beta}1$ was effective in bone formation and increased inductive effect of DFDB in guided bone regeneration technique. Inductive effect of DFDB was increased with $TGF-{\beta}1$.

• 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.23 no.2
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• pp.315-330
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• 1993

The ultimate goal of periodontal treatment has been to facilitate regeneration of diseased periodontal tissues, destroyed by inflammatory periodontal disease. Various implant materials have been used to restore the alveolar bone defects. Of the various materials, porous replamineform hydroxyapatite (PHA) has good biocompatibility when placed in a bone tissue, and maintains alveolar ridge for a long period. Decalcified freeze dried bone(DFDB) has been widely used in alveolar bone defects because of its conformity and high osteogenic potential. The purpose of this study was to evaluate the effects of PHA and DFDB on the regeneration of the alveolar bone between fresh extraction sockets and periodontally involved extraction sockets. Experimental periodontitis was induced by the ligation of orthodontic elastic threads after surgically creating periodontal defects on the premolars on the right side of 2 adult dogs for 8 weeks. Following the extraction of each tooth, PHA and DFDB were inserted in the extraction sockets. In control group 1, PHA was inserted in the fresh extraction sockets, and in control group 2, DFDB was inserted. In experimental group 1, PHA was inserted in the periodontally involved extraction sockets, and in experimental group 2, DFDB was inserted. After 20 weeks, the specimens were prepared and stained with Hematoxylin-Eosin stain for the light microscopic evaluation. The results of this study were as follows. 1. No inflammation associated with implant materials was evident in any of the groups. 2. DFDB was completely resorbed, PHA was remained in the extraction sockets in the control and experimental groups. 3. In control group 1 and experimental group 1, extraction sockets were not completely filled with new bone. However, original forms of alveolar crests were maintained in control group 2 and experimental group 2. 4. In control group 1 and exprimental group 1, PHA particles surrounded with many giant cells were well tolerated by the fibrous connective tissues in the coronal part of the socket, In the inferior part of the socket, PHA particles were incorporated into the new bone. In both control group 2 and experimental group 2, DFDB was replaced by newly remodeled bone. 5. No differences of degree of new bone formation were evident between control and experimental groups.

• 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.28 no.2
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• pp.219-237
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• 1998

Numerous bone graft materials have been used in Periodontics, in an attempt to reach the main goal of periodontal therapy, i.e. the regeneration of periodontal tissue lost due to destructive periodontal diseases. The present study investigates the effect of composite graft of DFDB and Calcium sulfate with and without Calcium sulfate barrier in Periodontal 1-wall intrabony defects in dogs. Following the initiation of general anesthesia by I.V. administration of 40mg/Kg of Pentobabital, second premolar was extracted and full thickness flap elevated. The crown portion of premolars was removed. Exposed root canals were sealed with Caviton and covered completely with flap. After the healing period of 8 weeks, the surgical sites were re-opened and 1-wall intrabony defects were created, and treated with flap operation alone(control group), with composit graft of 80% DFDB and 20% Calcium sulfate(Experimental group 1), with composite graft of DFDB and calcium sulfate with calcium sulfate membrane( Experimental group 2). Healing response was histologically observed after 8 weeks and the results were as follows : 1. New bone formation was 70 % in the control group, 93 % in the Experimental group I, 89 % in the Experimental group II. There was a no differences between Experimental groups. 2. New cementum formation was not significantly different between control and two Experimental groups. 3. The length of connective tissue adhesion was 30 % in the control, 7% in the Experimental group I and 11 % in the Experimental group II. 4. After 8weeks, calcium sulfate was completely resorbed, while DFDB particle remained. These results suggest that the use of composite graft of allogenic DFDB and Calcium sulfate with and without Calcium sulfate barrier in periodontal 1 wall intrabony defects have little effect on connective tissue adhesion, but has beneficial effect on new alveolar bone and new cementum formation, and prevent downgrowth of epithelium and connective tissue effectively.

• 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 Periodontal and Implant Science
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• v.27 no.2
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• pp.347-361
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• 1997

The present study investigates the effects of DFDB graft combined with Calcium sulfate membrane on the periodontal wound healing in dehiscence defects of dogs. Following the initiation of general anesthesia by I.V. administration of 30mg/kg of pentobarbital, first premolar was extracted and full-thickness flap was elevated from the second to the fourth premolar. The portion of premolars coronal to the alveolar crest was removed and mesial and distal roots separated to produce single rooted teeth. Exposed root canals were sealed with Caviton and covered completely with flaps sutured. Following the healing period of 12 weeks, the surgical sites were uncovered and $4{\times}4mm$ dehiscence defects were surgically created. Those defects with DFDB graft combined with Calcium sulfate membrane following root planing, were designated as test sites and those with flap surgery-only were designated as controls. 1. No foreign-body reaction or inflammation were observed in either groups. Calcium sulfate was completely resorbed in the test sites. 2. Significantly greater amounts of new cementum was observed in test sites compared with the controls. Significant amounts of functionally orientated collagens were observed in the test sites. 3. New bone formation was observed in significantly greater amounts in test sites. The results suggest that combined graft of DFDB and calcium sulfate is extremely biocompatible with a potential for new bone and cementum formation, and functional alignment of periodontal ligaments.

• Journal of Periodontal and Implant Science
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• v.28 no.2
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• pp.205-221
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• 1998

The present study invetigates the effects of root planing only(control group), DFDBA alone(test group 1) and combined use of DFDB and Dura mater(test group 2) in dehiscence defects in dogs. The results of 8weeks post-surgery by histological comparison between the three groups are as follows. 1. The contol group showed minimum regeneration of new cementum and new bone with limited migration of epitheilal cells, and healed by connective tissue attachment. 2. The test group 1 showed minimum regeneration of new cementum and new bone with limited migration of epitheilal cells, and healed by connective tissue attachment. 3. The test group 2 showed significant amount of the new cementum and new bone. 4. Both control and test groups healed without any observable root resorption and ankylosis. The above the results suggest that the use of resorbable Dura mater only does not improve the regeneration of new bone and periodontal ligament due to difficulties of space making, but the combined use with DFDB may be more effective.