• 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.

• Journal of Periodontal and Implant Science
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• v.28 no.1
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• pp.17-35
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• 1998

Chitosan, with a chemical structure similar to hyaluronic acid, has been implicated as a wound healing agent. The purpose of this research was to evaluate the effects of chitosan on the characteristics of periodontal ligament cells, calvaria cells and gingival fibroblasts and to define the effects of chitosan on bone formation in vitro. In control group, the cells were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% Fetal bovine serum, 100unit/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, chitosan($40{\mu}g/ml$) is added into the above culture condition. And then each group was characterized by examining the cell proliferation at 1,3,5,7,9,12,15 day, the amount of total protein synthesis, alkaline phosphatase activity at 3, 7 day and the ability to produce mineralized nodules of rat calvaria cell at 11 day. The results were as follows : 1. At early time both periodontal ligament cells and calvaria cells in chitosan-treated group proliferated more rapidly than in non-treated control group, but chitosan-treated group of periodontal ligament cells at 9 days and calvaria cells at 12days showed lower growth rate than control group. Gingival fibroblast in chitosan-treated group had lower growth rate than in control group but the difference was not statistically significant (P< 0.01).2. Both periodontal ligament cells and calvaria cells in chitosan-treated group showed much protein synthesis than in control group at 3 days, but showed fewer than in control group at 7 days. Amount of total protein synthesis of gingival fibroblast didn't have statistically significant difference among the two groups(P< 0.01). 3. At 3 and 7 days, alkaline phosphatase activity of periodontal ligament cells and calvaria cells was increased in chitosan-treated group, but at 7 days there was not statistically significant difference among the two groups of calvaria cells (P< 0.01). Alkaline phosphatase activity of gingival fibroblast didn't have statistically significant difference among the two groups(P<0.01). 4. Mineralized nodules in chitosan-treated group of rat calvaria cells were more than in control group. In summery, chitosan had an effect on the proliferation, protein systhesis, alkaline phosphatase activity of periodontal ligament cells and calvaria cells, and facilitated the formation of bone. It is thought that these effects can be used clinically in periodontal regeneration therapy.

• Toxicological Research
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• v.14 no.2
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• pp.163-169
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• 1998

Fluoride (F), over a narrow concentration range, increases bone formation. Aluminum (Ai) too is biphasic in its action on bone, being mitogenic at very low levels and inhibitory at higher levels. Both F and Al are present in finished drinking water where the chemical interaction of these two agents is well characterized. F and AI, given individually, accumulate preferentially in bone. In addition. in vivo studies have shown that F causes the co-accumulation of Al in bone. Thus, it was necessary to determine the interactive effect of these two agents on bone mitogenesis. Calvaria were obtained from neonatal CD-1 mice and cultured with various concentrations of F (0.05~19 ppm) as NaF, Al (2 ppb~2 ppm) as $AlCl_3$ , or F and Al for 3 days at $37^{\circ}C$ on a rotating roller drum. Alkaline phosphatase activity in calvaria and $\beta$-glucuronidase activity in culture medium were determined as a measures of bone turnover. Alkaline phosphatase activity in calvaria was significantly increased by F (0.05~2 ppm) treatment and $\beta$-glucuronidase activity was slightly increased in the culture medium of calvaria treated with 0.3 ppm Al. The combination of 19 ppm F and 0.3 ppm Al increased alkaline phosphatase activity in calvaria, but did not affect $\beta$-glucuronidase activity, suggesting the interactive effect of fluoride and aluminum on bone turnover.

• 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 the Korean Association of Oral and Maxillofacial Surgeons
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• v.44 no.2
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• pp.79-85
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• 2018

Objectives: The aim of this study was to evaluate the effects of herbal extracts on bone regeneration. Two known samples were screened. Materials and Methods: We previously established a rat calvaria defect model using a combination of collagen scaffold and herbal extracts. An 8 mm diameter trephine bur with a low-speed dental hand piece was used to create a circular calvaria defect. The experimental group was divided into 4 classifications: control, collagen matrix, Danshen with collagen, and Ge Gan with collagen. Animals in each group were sacrificed at 4, 6, 8, and 10 weeks after surgery, and bone regeneration ability was evaluated by histological examination. Results: Results revealed that both Danshen and Ge Gan extracts increased bone formation activity when used with collagen matrix. All groups showed almost the same histological findings until 6 weeks. However, after 6 weeks, bone formation activity proceeded differently in each group. In the experimental groups, new bone formation activity was found continuously up to 10 weeks. In the Danshen and Ge Gan groups, grafted materials were still present until 10 weeks after treatment, as evidenced by foreign body reactions showing multinucleated giant cells in chronic inflammatory vascular connective tissue. Conclusion: Histological analyses showed that Danshen and Ge Gan extractions increased bone formation activity when used in conjunction with collagen matrix.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 1996.12a
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• pp.47-47
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• 1996

• Journal of Periodontal and Implant Science
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• v.44 no.5
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• pp.251-258
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• 2014

Purpose: The preferred material for bone augmentation beyond the envelope of skeletal bone is the bone block graft, due to its dimensional stability. We evaluated the necessity of rigid fixation for the bone block graft, and compared the bone regeneration and volume maintenance associated with grafting using a synthetic hydroxyapatite block (HAB) and an autogenous bone block (ABB) without rigid fixation on rabbit calvaria over two different periods. Methods: Cylinder-shaped synthetic HAB and ABB were positioned without fixation on the rabbit calvarium (n=16). The animals were sacrificed at 4 or 8 weeks postoperatively, and the grafted materials were analyzed at each healing period using microcomputed tomography and histologic evaluation. Results: Integration of the graft and the recipient bed was observed in all specimens, although minor dislocation of the graft materials from the original position was evident in some specimens (six ABB and ten HAB samples). A tendency toward progressive bone resorption was observed in the grafted ABB but not in the grafted HAB, which maintained an intact appearance. In the HAB group, the area of new bone increased between 4 and 8 weeks postoperatively, but the difference was not statistically significant. Conclusions: The nonfixed HAB was successfully integrated into the recipient bed after both healing periods in the rabbit calvaria. In spite of limited bone formation activity in comparison to ABB, HAB may be a favorable substitute osteoconductive bone material.

• 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.27 no.4
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• pp.923-939
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• 1997

This study was performed to evaluate the effect of mixed culture of rat's calvaria cells and periodontal ligament cells on calcification. These cells have been known to do important role on the periodontal tissue regeneration, especially alveolar bone and cementum. Experimental groups were made which based on the different rate of rat's calvaria cells and periodontal ligament cells, and then these cells were cultured with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum, $50{\mu}g/ml$ ascorbic acid, and 10mM/ ml $Na-{\beta}-glycerophosphate$. Each group was characterized by examining the cell proliferation rate, amount of total protein synthesis, alkaline phosphatase activity, and the number of calcified nodules in vitro. In cell proliferation rate , the cells of control groups were cultured Dulbecco's Modified Eagle's Medium contained with 10 % fetal bovine serum. The results were as follows : 1. The cell proliferation rate in control groups decreased stastically significantly along with the decrease of the rate of bone cells at 7 day and 20 day(P < 0.01). 2. The cell proliferation rate in experimental groups decreased stastically significantly along with decrease of the rate of bone cells at 3 day and 14 day(P < 0.01). 3. The amount of total protein synthesis was significantly decreased along with decrease of the rate of bone cells at 3 day and 6 day(p < 0.01). 4. Alkaline phosphatase activity showed reverse time dependent pattern and was significantly decreased along with decrease of the rate of bone cells during the experimental periods (P < 0.01). 5. Calcified nodules were observed in group 1 (Rat calvaria cells alone) for the first time, and the number of calcified nodule decreased stastically significantly along with the decrease of the rate of bone cells at 12 day(P < 0.01). From the above results, When bone cells and periodontal ligament cells were mixed cultured, the cell proliferation rate was mostly dependent on the actual rate of bone cells and same pattern was showed in amount of total protein synthesis, alkalinephosphatase activity, and the number of calcified nodules. And the calcified nodule forming capacity of bone cells was inhibited by periodontal ligament cells

• Journal of Periodontal and Implant Science
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• v.27 no.1
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• pp.165-177
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• 1997

The purpose of this study was to perform on the biological activity of Magnolia and Zizyphi fructus extract mixtures on the wound healing of defected rat calvaria. For the determination of the mixture ratio of two extracts for oral administration, preliminary experiments were performed with the mixture combination of 2000 and $3000{\mu}g/ml$ of Magnolia extract, and also 20, 30, 200, 300, 2000 and $3000{\mu}g/ml$ of Zizyphi fructus extract, respectively and divided into 6 groups. The combination of extracts mixture were tested on the enhancing effect of cellular activity. The effect of the extracts mixture on the cellular activity was evaluated using MTT method and measured on the results with optical density by ELISA reader. The ability to tissue regeneration of the extracts mixture was performed by measuring new bone and new connective tissue regeneration on the 5mm defected rat calvaria for 1, 2 and 3 weeks after oral administration of 2 different dosages groups : 10:1(0.1g/kg) and 10:1(0.5g/kg). It was employed the same dosages of unsaponifiable fraction of Zea Mays L as positive controls. Each group of rat was sacrificed and en bloc section for histological examination. The effect on the cellular activity of each mixture ratio showed significantly higher in $2000{\mu}g/ml$ of Magnolia extract and $200{\mu}g/ml$ of Zizyphi fructus extract group to compare with other groups. These preliminary results showed that appropriate mixture ratio of two extracts was 10:1 of Magnolia and Zizyphi fructus extract. Histological examination on the activity of tissue regeneration of each group showed that 2weeks and 3weeks specimens of 0.5g/kg of 10:1 extract mixture of Magnolia and Ziziphi fructus administrated rat calvaria revealed significantly more osteoid and new bone formation of defected calvaria with unification of defected area than the specimens of any other negative and positive controls. Even though the specimen administrated the same dosages of unsaponifiable fraction of Zea Mays L, positive controls, showed the trend that they promote significantly the repair of calvarial defect, their bone reparative activities were less inductive than the same dosages of Magnolia and Ziziphi fructus extract mixture. These results implicated that the mixture of Magnolia and Zizyphi fructus extracts should be highly effective on the wound healing of bony defected site and might have potential possibilities as an useful drug to promote periodontal tissue regeneration.