• Maxillofacial Plastic and Reconstructive Surgery
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• v.19 no.3
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• pp.319-331
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• 1997

The purpose of this study was to evaluate the bone-forming capacity of the periosteum in calvaria of rats. The experiment was carried out in 49 rats. We exposed the calvaria and made 1㎝ diameter round full thickness defect at both sides of calvaria. In the left calvarial bone serving as control, the periosteum was removed after implantation of block, while in the right calvarial bone the periosteum remained intact as an experimental site. The histologic examination of bone response was performed after 1-, 2-, 4-, 6-,8-, 12-, 24-week implantation in calvaria of rats. We could observe the periosteal preservation favorably influenced the bone formation.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.20 no.2
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• pp.127-132
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• 1998

$Medpor^{(R)}$(porous polyethylene) Surgical Implants are used for the augmentation or restoration of bony contour in craniofacial defects. The purpose of this study is to evaluate the ingrowth of soft tissue and bone after application in calvaria of rats. The experiment was carried out in 60 rats. The reflected periosteum was resutured after implantation of $Medpor^{(R)}$ as a experimental site, while in the calvarial bone the reflected periosteum resutured without implantation as a control site. The histologic examination was performed after 1-, 2-, 4-, 8-, 12-, 24-weeks implantation in calvaria of rats. I concluded that there was abundant ingrowth of soft tissue and bone without any adverse tissue response and that it shows good stability.

• Journal of Periodontal and Implant Science
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• v.33 no.2
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• pp.193-213
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• 2003

The purpose of this study is to evaluate the phenomenon of attachment and spreading of the cultured rat　calvarial cell inoculated on their surface of different kinds of biodegradable membrane which had been used on tissue regeneration on periodontal defects by using scanning electron microscope. In this experiment 30 Sprague-Dawley male rats (mean BW 150gm) were used to harvest abundant number of cell in the short period. The rats were sacrificed by decapitatioan to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. Biodegradable barrier membrane were collected with collagen type, and were divided into 3 different kind of surface such as scattered, polarized and fine-net type as their surface texture. Microcover plate which usually used for cell culture was used as control for smooth surface. All the membrane were seeded with cultured calvarial cell on their surface. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. After the culture as designed time, all the membrane were washed with 0.1 M Phosphate Buffered saline and fuxed with 2.5% Glutaraldehyde. And all specimen were treated with $OsO_4$, and Tannic acid before drying the cell for coating the cell with gold. Scanning Electron Microscope was used to observation. The following results were obtained. I. During the whole period of experiment, the phenomenon of cell attachment and spreading were revealed similar pattern to compare with smooth surface culture plate and ordinary culture dish. 2. The shape of cell attachment and spreading on the surface of barrier membrane were observed no remarked difference pattern between smooth surface culture plate and ordinary culture dish. 3. The cytoplasmic process of cultured calvaria cell extent to the deep portion of barrier membrane like as their own proper shape. 4. There were no remarkable relationships between the degree of cultured cell spreading and surface structure of barrier membrane. 5. Slight starified layer of cultured calvaria cell were observed on the scattered type of resorbable membrane, Conclusively, this study thus suggest that cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of carrier for many cell which could be used as new tissue regeneration, and those tissue engeering technique may become an new method in the approach to the repair of bone defects.

• Journal of Periodontal and Implant Science
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• v.31 no.2
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• pp.389-400
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• 2001

Previous studies have demonstrated an increase in bone mass and density with the use of bisphosphonate in osteoporosis. This agent acts as an inhibitor of osteoclastic activity, and results in increase of net osteoblastic activity. Currently, it has been reported that bisphosphonate has direct effect on osteoblast. This study was designed to evaluate the effect of alendronate on bone regeneration in defect of rat calvaria. The animals used for these experiments were 48 male rats, over 6-8 weeks old. They were divided into three groups according to the dose of alendronate($MK-217^{(R)}$, Merck, USA) administered. After the calvarial defects were surgically created, the rats received a peritoneal alendronate(0.25mg/kg) in group I, a peritoneal alendronate(1.25mg/kg) in group II, and a peritoneal normal saline injection in the control group. Three and six weeks later, blood was sampled and evaluated for alkaline phosphatase activity. The animals were sacrificed for histological observation and histometric analysis of the level of bone formation. The alkaline phosphatase activity was similar in three groups at 3 weeks of experiment. The activity at 6 weeks increased more than twice, compared to 3 weeks, and was slightly higher in group I than the other two groups. In histological observation, all the groups at 3 weeks, osteoblast rimming and new bone formation were observed along the defect margin. At 6 weeks, the defect was almost closed with new and more mature bone, but new bone is thinner than original bone in the central portion of defect. In histometric analysis, group I and II at 3 weeks showed significantly greater new bone formation than the control, and all the groups at 6 weeks showed similar amount of bone formation. These result suggest that alendronate administration in the dose of 0.25mg/kg and 1.25mg/kg promote osseous regeneration.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.28 no.3
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• pp.205-215
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• 2002

The purpose of this study was to evaluate the tissue response in various bone grafting materials, especially xenogenous bone materials in vivo, compare of bone formation capacity of various bone grafting materials on rat skull defects and evaluate the effect of Hyaluronic acid on healing of human Demineralized Freezed Dried Bone allogenous graft (DFDBA) materials in rat calvarial defects. 30 Sprague-Dawly rats were divided into 4 groups. $7{\times}7mm$ size bony defect were artificially prepared in the calvaria (both parietal bone) of all 30 rats and follwed group grafting of autogenous bone graft on right side and allogenic DFDBA on left side bone graft (rat DFDB) in 15 control group, but in 15 experimental group, xenograft (human DFDB) on left side, hyaluronic acid treated with xenograft on right side. Sequential sacrifices was performed at 1, 2, 4, 6, 8 weeks of experiment. These specimens were stained with H&E and MT stain, and then histologic analysis under light microscope was carried out. There were inflammatory reaction in all graft material during early stage. Autogenous and Allogenous DFDBA graft group observed inflammatory reaction at 1 week. Xenograft group persistant inflammatory reaction until 4 weeks, but in HA treated xenograft group inflammatory reaction was decreased at 2 weeks. Osteoblastic activity in control group was begun at 2 week, xenograft group was delayed at 6 weeks, however HA treated xenograft group was begun at 4 weeks. At 2 week, mild osteoclastic activity were observed in all xenograft group not in concerned to HA, but there was no difference each group after 4 weeks. There are most activated angiogenesis around graft mateirals in xenograft group at 2 weeks, but in HA treated xenograft group, decreased angiogenesis was observed at same time. Bone formation and bone maturation of xenograft group, there was no difference in HA treatment, was less than control group. Fibrosis around xenograft materials were observed until 6 weeks, there was no difference between xenograft and HA treated groups.

• Journal of Periodontal and Implant Science
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• v.47 no.2
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• pp.77-85
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• 2017

Purpose: Guided bone regeneration (GBR) is the most widely used technique to regenerate and augment bones. Even though augmented bones (ABs) have been examined histologically in many studies, few studies have been conducted to examine the biological potential of these bones and the healing dynamics following their use. Moreover, whether the bone obtained from the GBR procedure possesses the same functions as the existing autogenous bone is uncertain. In particular, little attention has been paid to the regenerative ability of GBR bone. Therefore, the present study histologically evaluated the regenerative capacity of AB in the occlusive space of a rat guided bone augmentation (GBA) model. Methods: The calvaria of 30 rats were exposed, and plastic caps were placed on the right of the calvaria in 10 of the 30 rats. After a 12-week healing phase, critical-sized calvarial bone defects (diameter: 5.0 mm) were trephined into the dorsal parietal bone on the left of the calvaria. Bone particles were harvested from the AB or the cortical bone (CB) using a bone scraper and transplanted into the critical defects. Results: The newly generated bone at the defects' edge was evaluated using micro-computed tomography (micro-CT) and histological sections. In the micro-CT analysis, the radiopacity in both the augmented and the CB groups remained high throughout the observational period. In the histological analysis, the closure rate of the CB was significantly higher than in the AB group. The numbers of cells positive for runt-related transcription factor 2 (Runx2) and tartrate-resistant acid phosphatase (TRAP) in the AB group were larger than in the CB group. Conclusions: The regenerative capacity of AB in the occlusive space of the rat GBA model was confirmed. Within the limitations of this study, the regenerative ability of the AB particulate transplant was inferior to that of the CB particulate transplant.

• Korean Journal of Veterinary Research
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• v.60 no.3
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• pp.145-153
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• 2020

This study aimed to evaluate the effects of a bioabsorbable bone hemostatic agent comprising poly (ethylene glycol-propylene glycol) copolymers (PEG-PPG) on hemostasis and osteogenesis. Bilateral 3 mm diameter calvarial defects were created in 99 male Sprague-Dawley rats. The defects were filled with PEG-PPG or bone wax. The defects of control group were left unfilled. Virtual autopsy was performed to evaluate bioabsorption. The calvaria were subjected to x-ray microtomography (microCT) and histological examination. Bone volume fraction (BV/TV) and bone mineral density (BMD) were measured using microCT; furthermore, white blood cell count and histological examination were performed. After application of PEG-PPG and bone wax, immediate hemostasis was achieved. Autopsy revealed that PEG-PPG disappeared within 48 h at the application site; in contrast, bone wax remained until 12 weeks. The PEG-PPG and control groups showed significantly more osteogenesis than the bone wax group with respect to BV/TV and BMD at 3, 6, and 12 weeks (p < 0.05). Histology revealed that the bone wax group exhibited little bone formation with inflammation. In contrast, PEG-PPG and control groups showed significantly more qualitative osteogenesis than the bone wax group (p < 0.01). In conclusion, PEG-PPG showed immediate hemostasis and was absorbed to allow progressive osteogenesis.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.21 no.1
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• pp.1-12
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• 1999

The purpose of this study was to evaluate the healing process of the calvarial defect filled with hydroxylapatite(HA) and $TGF-{\beta}$ in Rat. 72 Sprague-Dawly rats were divided into 3 groups, control and two experimental groups. Bony defect were artificially prepared in the calvaria of all 72 rats and followed by implantation of HA (experimental group of 24 rats) and HA+$TGF-{\beta}$(another experimental group of 24 rats) into the defects. Sequential sacrifice was performed at 1, 2, 4, 6, 8, 12 weeks of experiment. Obtained specimen was stained with Hematoxylin and Eosin, Masson's Trichrome and Immunohistochemistry. The results were as follows, 1. Granulation tissue was prominent on control group in 1 and 2 weeks. Bony defects were filled with dense fibrous tissue through the whole experimental period and osteoinduction could not be observed in all groups. 2. Inflammatory cell infiltration was prominent on control group in 1 and 2 weeks and osteoclastic activity was high in HA implanted experimental group at 1 and 2 weeks. 3. Inflammatory cell infiltration was less and maturation of fibrous tissue could be found on HA+$TGF-{\beta}$ implanted experimental group at 1 and 2 weeks. 4. Osteoconduction activity was high in HA+$TGF-{\beta}$ implanted experimental group at 2 and 4 weeks but there was no difference after 6 weeks among 3 groups. 5. In grafted site of HA+$TGF-{\beta}$ implanted group, osteonectin expression was slightly increased from 1 week to 6 weeks. In the host site, it was increased from 1 to 4weeks. 6. In grafted site of HA+$TGF-{\beta}$ implanted group, osteocalcin expression was high at 4 weeks. In the host site, we could find the difference among 3 groups. From above results, the HA with mixture of $TGF-{\beta}$ has the potentiality of promoting bone formation in the bony defect area in the rat.

• 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.49 no.1
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• pp.2-13
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• 2019

Purpose: The aim of this study was to conduct a histologic evaluation of irradiated calvarial defects in rats 4 weeks after applying fibroblast growth factor-2 (FGF-2) with hyaluronan or biphasic calcium phosphate (BCP) block in the presence or absence of adjunctive hyperbaric oxygen (HBO) therapy. Methods: Twenty rats were divided into HBO and non-HBO (NHBO) groups, each of which was divided into FGF-2 and BCP-block subgroups according to the grafted material. Localized radiation with a single 12-Gy dose was applied to the calvaria of rats to simulate radiotherapy. Four weeks after applying this radiation, 2 symmetrical circular defects with a diameter of 6 mm were created in the parietal bones of each animal. The right-side defect was filled with the materials mentioned above and the left-side defect was not filled (as a control). All defects were covered with a resorbable barrier membrane. During 4 weeks of healing, 1 hour of HBO therapy was applied to the rats in the HBO groups 5 times a week. The rats were then killed, and the calvarial specimens were harvested for radiographic and histologic analyses. Results: New bone formation was greatest in the FGF-2 subgroup, and improvement was not found in the BCP subgroup. HBO seemed to have a minimal effect on new bone formation. There was tendency for more angiogenesis in the HBO groups than the NHBO groups, but the group with HBO and FGF-2 did not show significantly better outcomes than the HBO-only group or the NHBO group with FGF-2. Conclusions: HBO exerted beneficial effects on angiogenesis in calvarial defects of irradiated rats over a 4-week healing period, but it appeared to have minimal effects on bone regeneration. FGF-2 seemed to enhance new bone formation and angiogenesis, but its efficacy appeared to be reduced when HBO was applied.