• Journal of Veterinary Clinics
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• v.20 no.4
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• pp.482-488
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• 2003

Transplanted cortical bone grafts of freeze-dried bones also function as sustaining for defected bones, however, it has less strength and is fragile without rehydration. In this study, strength and stiffness of freeze-dried bone from bovine cortical bones were evaluated by three point bending test according to different time frames such as rehydration times of 0.5, 3, 6, 12 and 24 hrs in electrolyte solution and was compared with those of frozen bones. The strength and stiffness of frozen bone were $264.4\pm36.7$ MPa, $17.0\pm1.5$ GPa, respectively. The strength and stiffness of freeze-dried bone which fat was removed by treatments of chloroform-methanol solutions for 6 days, then was freeze-dried at $-80^{\circ}C$ and sterilized with ethylene oxide gas, were $224.9\pm27.6$ MPa, $19.2\pm2.8$ GPa, respectively. The strength and stiffness of feeze-dried bone were decreased 15.0% and increased 13.2% than these of frozen bone, respectively. The strength and stiffness of freeze-dried bone rehydrated for 6 hrs were restored to 96.0% strength and 99.2% stiffness of frozen bone. The rehydration time of freeze-dried bone which had the highest strength and stiffness was six hours and three hours, respectively. The results of the mathematica program for the variation of the strength and stiffness showed 3 hours and 30 minutes of rehydration time in electrolyte solution for the best condition in the strength and stiffness which was adequate to treat freeze-dried cortical bone.

• Journal of Periodontal and Implant Science
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• v.40 no.3
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• pp.132-138
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• 2010

Purpose: The purpose of this study was to compare the bone regeneration effects of cortical, cancellous, and cortico-cancellous human bone substitutes on calvarial defects of rabbits. Methods: Four 8-mm diameter calvarial defects were created in each of nine New Zealand white rabbits. Freeze-dried cortical bone, freeze-dried cortico-cancellous bone, and demineralized bone matrix with freeze-dried cancellous bone were inserted into the defects, while the non-grafted defect was regarded as the control. After 4, 8, and 12 weeks of healing, the experimental animals were euthanized for specimen preparation. Micro-computed tomography (micro-CT) was performed to calculate the percent bone volume. After histological evaluation, histomorphometric analysis was performed to quantify new bone formation. Results: In micro-CT evaluation, freeze-dried cortico-cancellous human bone showed the highest percent bone volume value among the experimental groups at week 4. At week 8 and week 12, freeze-dried cortical human bone showed the highest percent bone volume value among the experimental groups. In histologic evaluation, at week 4, freeze-dried cortico-cancellous human bone showed more prominent osteoid tissue than any other group. New bone formation was increased in all of the experimental groups at week 8 and 12. Histomorphometric data showed that freeze-dried cortico-cancellous human bone showed a significantly higher new bone formation percentile value than any other experimental group at week 4. At week 8, freeze-dried cortical human bone showed the highest value, of which a significant difference existed between freeze-dried cortical human bone and demineralized bone matrix with freeze-dried cancellous human bone. At week 12, there were no significant differences among the experimental groups. Conclusions: Freeze-dried cortico-cancellous human bone showed swift new bone formation at the 4-week healing phase, whereas there was less difference in new bone formation among the experimental groups in the following healing phases.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.18 no.1
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• pp.164-174
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• 1996

In order to develop the allogeneic bone implants instead of autogenous bone grafts for maxillofacial reconstruction, undemineralized freeze-dried human bone was processed. The freeze-dried human bone was implanted into the cranial and mandibular defects of the rabbits. The implants were evaluated clinically, roentgenographically and histomophometrically. And immunohistochemical evaluation of the implants was performed on the rat. The results were as follows : 1. When compared with control defects of $0.8{\times}0.8\;cm$, the implants on the rabbit defects displayed complete osseous bridging clinically and roentgenographically. Histomophometrically a minimal inflammatory cell infiltrate was present but the defects healed well clinically. 2. When compared with control grafts, the freeze-dried implants on the rat muscle displayed decreased antigenicity by immunohistochemical evaluation, due to freeze-drying process. 3. Undemineralized freeze-dried human bone in this study can be preserved as a bank bone in this study and seems to be applicable for clinical allogeneic bone grafts.

• Journal of Veterinary Clinics
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• v.15 no.2
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• pp.442-449
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• 1998

Freeze-dried cortical bones of the goat were transplanted to the experimental fibular defect of 10 dogs for valuating the possibility of xenogeneic bone implantation and the specificity of BM(Bone Morphogenetic Protein). The . freeze-dried cortical bone eliminated antigens and defatted with chloroform and methanol were freeze-dried at $-80{\circ}C$ for preservation of BMP and then sterilized with 50 gas and storaged in room temperature. Ten freeze-dried cortical implants of the goat were transplanted in experimentally defected regions of bilateral fibula of 5 dogs in clinically normal. The transplanted region had been radiographed for observing state of bone union and BALPOone Alkaline Phosphatase) in the serum of the host was measured for valuating activity of oteoblast per 2 week-interval after transplant procedures. New bone formation had been observed early in one of ten regions around implants about the same time as autoimplant regions. It was incorporated with its host bone during 4-12 weeks after transplantation. In another 2 cases of 2 dogs, new bone formation and absorption of implant had been observed from 4 weeks but they were not incorporated completely until 20 weeks. The rest of the freeze-dried bone implants, 7 cases of 4 dogs had not been observed new bone formation nor absorption of implants. The freeze-drying method for implants means to not influence bone incorporation. Although less of union percentages the union form of this experiment were similar to alloimplantation and it may mean to block immunity reaction that disturbs the bone induction by BMP. It demonsknted that the possibility of the xenogenous bone implantation is recognized by reason of the low specificity of BMP between goat and dog.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.17 no.2
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• pp.137-152
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• 1995

This study was designed to evaluate the bone formation capability of the bone substitute when compared with autogenic bone, freeze-dried demineralized allogeneic bone and bioglass into parietal bone of the rats. We made the parietal bone defects in $7{\times}7mm$ size on rats and has performed the bone graft in each experimental groups. Postoperatively 1, 2, 4, 6, 8, weeks, each specimen stained with H & E, Masson's trichrome methods. We evaluated the osteogensis capability in each groups. The result were as follow : 1. Inflammatory cell infiltration approached at 1 week and disappeared at 4 weeks in all experimental group, expecially severe in freeze-dried demineralized allogeneic bone group. 2. New capillry proliferation was increased in autogeneic bone graft group than any other groups and was increased till 2 weeks and decreased in freeze-dried demineralized allogeneic bone group and was few in bioglass group. 3. Osteoblastic activity increased in autogeneic bone and freeze-dried demineralized allogeneic bone groups till 4 weeks, and decreased in 6 weeks which no difference between these groups. But, few occurred in bioglass group till 6 weeks. 4. Initial osteoclastic activity was prominent in freeze-dried demineralized allogeneic bone group and few in autogeneic bone group. 5. New bone formation bega at 1 week in autograft and freeze-dried demineralized allogenic bone groups, but, mild new bone formation at 8 weeks in bioglass.

• Journal of Periodontal and Implant Science
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• v.34 no.1
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• pp.163-175
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• 2004

This study was performed to evaluate bone formation in the calvaria of rabbit by the concept of guided bone regeneration with titanium mesh membrane and demineralized freeze-dried bone. The animal was sacrificed at 2 weeks, 4 weeks, 8 weeks, and 12 weeks after the surgery. Non-decalcified specimens were processed for histologic analysis. 1. The titanium mesh but the biocompatibility was excellent the cell-occlusiveness was feeble. 2. The cell-occlusiveness was feeble and also the soft tissue growth of the upper part of the newly-formed bone after operating was excellent in early stage. 3. The maintenance ability of the space for the GBR very was excellent. 4. The titanium mesh the tissue-integration was superior the wound fixation ability excellent. 5. The demineralized freeze-dried bone did not promote the bone regeneration. 6. With the lapse of time, formation quantity of the bone some it increased, it increased quantity very it was feeble. Within the above results, the titanium mesh for the guided bone regeneration was excellent, the dεmineralized freeze-dried bone confirmed does not promote bone regeneration.

• Korean Journal of Veterinary Research
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• v.36 no.2
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• pp.495-511
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• 1996

To investigate the effectiveness of the freeze-dried allografts and fibrin glue in bone grafts, the status of new bone formation and union of the grafted bone were observed in three types of grafting bones; autogenic bone(AT), allogenic bone(AL), and allogenic bone particles mixed with fibrin glue(FG). These were transplanted into non-union fracture model of 7 adult dogs with 2cm defect made in the proximal metaphysis of both fibulae. The autogenic and allogenic grafting bones had been treated by a modified freeze-dried method. The serial radiogram were observed the repair process of grafted bones biweekly until 17 or 21 weeks after transplantation and the observation of histological aspects, tetracycline double labeling and microradiography in the grafted bones were undertaken at 17 or 21 weeks after transplantation. The incorporation of bone minerals to the non-union fracture models were accomplished in 4 of 5 cases grafted with AL and in 2 of 4 cases grafted with AT. None of 5 cases grafted with FG were incorporated. The process of new bone formation and resorption in the grafted bones were observed three types; resorption of the grafted bones after newbone formation(type A) in 4 cases, new bone formation after resorption(type B) in 2 cases and complete or incomplete resorption without new bone formation(type C) in 8 cases. The modified freeze-dried method used in this study contributed to inhibite the rejection in allogenic grafts but the union period of the grafted freeze-dried bone was more prolonged than that of fresh autografts. Fibrin glue did not contribute to induce a new bone formation ofbone grafts.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.13 no.3
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• pp.265-277
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• 1991

In this study, the healing changes of the implanted bone and its surrounding tissues were examined on the histopathologic basis following implantation of the freeze - dried and radiation - sterilized allogeneic bone in Rectus abdominicus of the rat. This study was performed to see the tissue recations after implantation of the freeze - dried and radiation - sterilized allogeneic bone and whether osteogenesis or osteo - induction or osteo - conduction is happened. And the results were as follows : 1. The shape of the implanted allogeneic bone of the 1, 2 - week group specimen was similar to that of normal bone in light - microscopic finding and the atrophy of cellular organells was found in trans - mission electron - microscopic finding. 2. The implanted allogeneic bone was surrounded with the dense fibroconnective tissues, and infiltration of the chronic inflammatory cells gradually became increased. 3. Hyaline degeneration was observed in the surrounding tissue at the 3, 4, 6 - week group specimen. 4. Light - microscopically the resorption of implanted bone became prominent after 4 - week group and the necrosis of allogeneic bone implant became severe with loss of cell components in lacuna. 5. Electron - microscopically, the osteoclast - like cells ere fond after, 2 - week group. It is summarized that the osteo - conduction potential of the bone is remained just after implanting the freeze - dried and radiation - sterilized allogeneic bone on Rectus abdominicus of the rat, but gradually it disappeared with the gradual increse of chronic inflammatory reaction and osteoclastic activity. So it is suggested that the antigenicity of the freeze - dried and radiation - sterilized bone is remained and it has little osteo - conductive activity when it is implanted in the muscle.

• Journal of Periodontal and Implant Science
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• v.36 no.2
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• pp.289-303
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• 2006

This study was performed to evaluate the effect of bone graft materials including demineralized freeze-dried bone, freeze-dried bone, deproteinized bovine bone on space-making capacity and bone formation in guided bone regeneration with titanium reinforced ePTFE membrane(TR-ePTFE). Adult male rabbits(mean BW 2kg) were used in this study. Intramarrow penetration defects were surgically created with round bur on calvaria of rabbits. TR-ePTFE membrane was adapted to calvarial defect and bone graft materials were placed. Animals were sacrificed at 2, 8, 12 weeks after surgery. Non-decalcified specimens were processed for histologic analysis and prepared with Villaneuva bone stain. The results of this study were as follows: 1. TR-ePTFE membrane was biocompatible and capable of maintaining the space-making. 2. Tissue integration was not good at TR-ePTFE membrane. Fixation was not enough. so, wound stabilization was not good. 3. In animals using deproteinized bovine bone, demineralized freeze-dried bone, bone formation was little. 4. In animals using freeze-dried bone, bone formation was better. Within the above results, bone formation may be inhibited when wound stabilizafion was not good.

• Journal of Veterinary Clinics
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• v.20 no.1
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• pp.91-95
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• 2003

It has been known that periods of absorption varies allografts or xenografts of transplantations of freeze-dried cortical bone(FDCB). In this study changes of absorption of FDCB in xenograft transplantations were evaluated according to extracted time with chloroform-methanol solution(CM sol.). The FDCB from pig was removed soft tissue by surgical knife. Fat of the FDCB was removed with treatments of CM sol. for 2, 6, and 10 days, then the treated FDCB was freeze-dried at $-80^{\circ}C$ and sterilized with ethylene oxide gas. The FDCB was transplanted to fifteen millimeter artificial-defected regions of 6 dogs on fibular diaphyses. This was biweekly examined by radiograph for 18 weeks. In result new bone formation with FDCB treated for 6 days was higher than the other bones treated for 2 and 10 days. Duration of absorption with FDCB treated for 6 days was longer than the others. The remain with FDCB treated for 10 days was more than the others.