• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제31권3호
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• pp.228-238
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• 2005

Purpose: Several cryoprotectants are in use to help the survival of cells during cryopreservation of bone in maxillofacial region. Among them, $Me_2SO$(dimethyl sulfoxide), EG(ethylene glycol), sucrose were used for experimentally created defects with accompanying cryopreserved bone graft in the rabbit model. The aim of this study is to analyze the effect of above mentioned agents on bone formation using histologic and histomorphometrical methods, thus to provide experimental support for clinical application of these agents. Materials and methods: Nine rabbits were used as experimental animals. Surgical defects were created on the distal femoral heads and mesial tibial heads of each animal using trephine drill(5mm diameter and 5mm length). The harvested bones were cryopreserved in $-80^{\circ}C$ refrigerator for one week. The defects were filled with cryopreserved bone with cryoprotectants as experimental groups and cryopreserved bone without cryoprotectant as control. Then, the animals were sacrificed at 1, 2, and 3 weeks after surgery. With Goldner's modified Masson trichrome staining and semiautomatic image analysis system, we observed the change of the cells and bone formation. Results: After bone graft, bone formation and active remodeling process were examined in all experimental groups and the control. But the intensity of such activities of the control were somewhat weaker than that of the experiments. Especially $Me_2SO$+sucrose group was the best in bone formation and bone remodeling. $Me_2SO$ group was more than that of EG group in bone fomation. Sucrose seems to be helpful in survival of the bone cell. Histologic findings showed superior bony quantity and quality in experimental groups than that in control. Conclusions: The data from this study provides the basis for future studies for evaluating the effect of cryoprotectants in the cryopreservation of bone and clinical study for predictable use of these agents.

• Journal of Korean Neurosurgical Society
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• 제57권4호
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• pp.242-249
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• 2015

Objective : The timing of cranioplasty and method of bone flap storage are known risk factors of non-union and resorption of bone flaps. In this animal experimental study, we evaluated the efficacy of cranioplasty using frozen autologous bone flap, and examined whether the timing of cranioplasty after craniectomy affects bone fusion and new bone formation. Methods : Total 8 rabbits (male, older than 16 weeks) were divided into two groups of early cranioplasty group (EG, 4 rabbits) and delayed cranioplasty group (DG, 4 rabbits). The rabbits of each group were performed cranioplasty via frozen autologous bone flaps 4 weeks (EG) and 8 weeks (DG) after craniectomy. In order to obtain control data, the cranioplasty immediate after craniectomy were made on the contralateral cranial bone of the rabbits (control group, CG). The bone fusion and new bone formation were evaluated by micro-CT scan and histological examination 8 weeks after cranioplasty on both groups. Results : In the micro-CT scans, the mean values of the volume and the surface of new bone were $50.13{\pm}7.18mm^3$ and $706.23{\pm}77.26mm^2$ in EG, $53.78{\pm}10.86mm^3$ and $726.60{\pm}170.99mm^2$ in DG, and $31.51{\pm}12.84mm^3$ and $436.65{\pm}132.24mm^2$ in CG. In the statistical results, significant differences were shown between EG and CG and between DG and CG (volume : p=0.028 and surface : p=0.008). The histological results confirmed new bone formation in all rabbits. Conclusion : We observed new bone formation on all the frozen autologous bone flaps that was stored within 8 weeks. The timing of cranioplasty may showed no difference of degree of new bone formation. Not only the healing period after cranioplasty but the time interval from craniectomy to cranioplasty could affect the new bone formation.

• Journal of Periodontal and Implant Science
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• 제34권2호
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• pp.411-424
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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. Two different titanium meshes with varying number (353, 565) of pore were utilized in the study. Two surgical sites(T353, T565) were evaluated about whether or not the number of pore may have effect on the bone formation. The animal was sacrificed at 10days, 3 weeks, 6weeks, and 8 weeks after the surgery. Non-decalcified specimens were processed for histologic analysis. 1. Titanium mesh was biocompatible and capable of maintaining the spacemaking. 2. At 3 weeks, 6 weeks, and 8 weeks after GBR procedure, bone formation was more in the T353 site than in the T565 site. 3. Soft tissue layer above the regenerated bone was better developed in the T565 site. 4. There was no difference between two membranes in bone maturity with time. Within the above results, titanium mesh with lesser pore in number might be recommended for the early bone formation.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제38권4호
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• pp.221-230
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• 2012

Objectives: This study sought to evaluate the efficacy of collagen graft materials, as compared to other graft materials, for use in healing calvarial defects in rabbits. Materials and Methods: Ten mm diameter calvarial defects were made in ten rabbits. The rabbits were then divided into 4 groups: control, autogenous bone graft, SureOss graft, and Teruplug graft. Bone regeneration was evaluated using histological and radiographic methods. Results: Based on visual examination, no distinct healing profile was observed. At 4 weeks after treatment, histological analysis showed there was no bone regeneration in the control group; however, at 8 weeks after treatment, new bone formation was observed around the margin of the defective sites. In the autogenous bone graft group, new bone formation was observed at 4 weeks after treatment and mature bone was detected around the grafted bone after 8 weeks. In the SureOss graft group, at 4 weeks after treatment, acute inflammatory and multinuclear cells were noted around the grafted materials; at 8 weeks after treatment, a decrease in graft materials coupled with new bone formation were observed at the defective sites. In the Teruplug graft group, new bone formation was detected surrounding the bone margin and without signs of inflammation. There were statistically significant differences observed between the graft and control group in terms of bone density as evidenced by radiographic analysis using computed tomography (P<0.05), particularly for the autogenous bone graft group (P<0.001). Conclusion: These results suggested that autogenous bone, SureOss and Teruplug have the ability to induce bone regeneration as compared to an untreated control group. The osteogenic potential of Teruplug was observed to be lower than that of autogenous bone, but similar to that of SureOss.

• 대한수의학회지
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• 제39권6호
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• pp.1197-1209
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• 1999

Twenty-one dogs(male 11 heads, female 10 heads) which were about 16 months ($16.3{\pm}3.5$) old and 10kg($10.1{\pm}2.0$) body weight, were allotted randomly into four groups as follows. Group I consisted of five dogs whose muscles were operated for sham muscle injuries. Group II consisted of seven dogs treated for cystic duct obstruction. Group III consisted of five dogs treated for the union fracture model. Group IV consisted of five dogs treated for the non-union fracture model. Radiographical and histological observations were carried out to determine bone alkaline phosphatase(BALP) and total alkaline phosphatase(TALP) values of each group for 20 weeks after the treatments with the condition of new bone formation. And also the applicability of percentage of BALP values to TALP (B/T) was studied after BALP was compared respectively with TALP. The level of TALP was increased without any relation to bone formation in group II, and all levels of BALP and B/T were increased in group III. The mean of B/T was high in statistical significance, due to varied levels of B/T and BALP. The changes of rates of B/T were significantly increased only in the case of the active new bone formation in group III, union fracture model. It was recognized that the mean values of B/T were statistical significant of the high applicability of the B/T ratio as an index of bone formation.

• 한국재료학회:학술대회논문집
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• 한국재료학회 2009년도 추계학술발표대회
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• pp.40.2-40.2
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• 2009

Using the favorable resorption behavior of amorphous Calcium phosphate (CaP) we fabricated a gelatin basednano fibrous mat by electrospinning for using as a fast healing patch for minorbone defects. Bone is predominantly formed by an inorganic phase of nano-crystalline HAp materials and nano fibrous protein material of collagen. The osteoblast cells, which are the bone formation cells and are key to the new bone formation, receive these materials to form new bone. Taking these considerations we make a new nano fibrous mat of amorphous CaP and gelatin, which is derived from collagen itself. A polymer carrier of poly caprolactone(PCL) was used in the system to stabilize the materials in biological condition. The electrospinning conditions were optimized for smooth mat without any droplet formation. The fabricated mat was characterized for its morphologyby SEM. Mechanical properties like tensile strength was evaluated. To investigate the bio-compatibility we performed the MTT assay and investigated its resorption behavior and apatite formation behavior by SBF immersion.

• Journal of Periodontal and Implant Science
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• 제36권2호
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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 the Korean Association of Oral and Maxillofacial Surgeons
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• 제28권3호
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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.

• Imaging Science in Dentistry
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• 제40권3호
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• pp.137-142
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• 2010

Purpose : This study was performed to evaluate the diagnostic ability of ultrasonography in detection of bone defects and new bone formation. Materials and Methods : Experimental bony defects were prepared on the parietal bone samples acquired from 3.5 kg New Zealand male rabbits. The defects were evaluated using ultrasonography and CBCT, and examined histologically at interval of 1, 3, 6, and 8 weeks. Results : Ultrasonograph demonstrated hyperechogenicity in the defect area at 3 weeks and broadened hyperechogenicity from the margin of bone defect at 6 and 8 weeks due to new bone formation. On the CBCT images, new bone formation was first observed at 3 weeks around the margin of the defect, and showed gradually increase at 6 and 8 weeks. Histologic findings revealed existence of the fibroblasts and fibrous connective tissue with abundant capillary vessels only at 1 week, but osteoid tissue and newly formed trabecular bone at 3 weeks. Bone remodeling in the defect area was observed at 6 weeks and increased calcification and dense trabecular bone formation was observed at 8 weeks. Conclusions : Ultrasonograph proved to be a very useful diagnostic tool in detecting the bony defect and new bone formation. Additionally, ultrasonography provided valuable information regarding the blood supply around the defect area.

• BMB Reports
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• 제53권9호
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• pp.472-477
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• 2020

Osteoclasts are hematopoietic-derived cells that resorb bone. They are required to maintain proper bone homeostasis and skeletal strength. Although osteoclast differentiation depends on receptor activator of NF-κB ligand (RANKL) stimulation, additional molecules further contribute to osteoclast maturation. Here, we demonstrate that protocadherin-7 (Pcdh7) regulates formation of multinucleated osteoclasts and contributes to maintenance of bone homeostasis. We found that Pcdh7 expression is induced by RANKL stimulation, and that RNAi-mediated knockdown of Pcdh7 resulted in impaired formation of osteoclasts. We generated Pcdh7-deficient mice and found increased bone mass due to decreased bone resorption but without any defect in bone formation. Using an in vitro culture system, it was revealed that formation of multinucleated osteoclasts is impaired in Pcdh7-deficient cultures, while no apparent defects were observed in differentiation and function of Pcdh7-deficient osteoblasts. Taken together, these results reveal an osteoclast cell-intrinsic role for Pcdh7 in maintaining bone homeostasis.