• Journal of Periodontal and Implant Science
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• 제33권2호
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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.

• 대한수의학회지
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• 제50권3호
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• pp.171-177
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• 2010

The purpose of this study was to evaluate the osteogenic capacity of water extract of danshen (Salvia miltiorrhiza Bunge). We have established in rat critical-sized calvarial defect model using the combination with collagen scaffold and danshen hydrophilic extract. All rats were extinguished at 8 weeks after bone graft surgery, and the bone regeneration ability of bone grafting sides was evaluated by plain radiography and micro-CT. These results revealed water extract of danshen had the potential to promote osteogenesis especially continuous oral administration with local treatment compared to one-shot local treatment. This compound may provide a new alternative agent for growth factors to promote bone healing and bone regeneration. In conclusion, these results suggest that danshen hydrophilic extract have the potential to promote osteogenesis in bone defects. Further studies about fusion technology with salvianolic acid B, peptides, growth factors, and scaffolds using of the combination of tissue engineering, cell engineering and mechanical engineering are needed.

• Journal of Periodontal and Implant Science
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• 제38권4호
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• pp.595-602
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• 2008

Purpose: Ti-6Al-4V alloy is widely used as an implant material because of its good biocompatibility and good mechanical property compared with commercial pure titanium. Otherwise, toxicity of aluminum and vanadium in vivo has been reported. Ti-8Ta-3Nb alloy is recently developed in the R&D Center for Ti and Special Alloys and it was reported that this alloy has high mechanical strength, no cytotoxicity and similar biocompatibility to commercial pure titanium, but many studies are needed for its clinical use. In these experiment, we carried out different surface treatment on each Ti-8Ta-3Nb alloy disks, then cultured cell on it and assessed biological response. Materials and Methods: cpTi, Ti-6Al-4V, Ti-8Ta-3Nb alloy disks were prepared and carried out sandblasting and acid etching (SLA) or alkali-heat treatment (AH) on the Ti-8Ta-3Nb alloy disks. We cultured primary rat calvarial cells on each surface and assessed early cell attachment and proliferation by scanning electron microscopy, cell proliferation, alkaline phosphatase activity. Result: The rates of cell proliferation on the cpTi, Ti-8Ta-3Nb AH disks were higher than others (p<0.05) and alkaline phosphatase activity was significantly enhanced on the Ti-STa-8Nb AH disks(p<0.05). Conclusion: Most favorable cell response was shown on the Ti-8Ta-3Nb AH surfaces. It is supposed that alkali-heat treatment of the Ti-8Ta-3Nb alloy could be induced earlier bone healing and osseointegration than smooth surface.

• Journal of Periodontal and Implant Science
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• 제35권2호
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• pp.277-288
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• 2005

There is a potential role of collagenase-3 in alveolar bone loss and periodontal disease progression, we need to develope or find chemotherapeutic drugs or herbal agents which may regulate the expression of MMP-13. Ginseng saponin, one of the major components of Korea ginseng(panax ginseng) root, has many various biologic effects, such as cytotoxic effect, tumoricidal effects, cytokine regulations, and protein biosynthesis effect. The purpose of this study was to determine the effects of Korea red ginseng saponin on MMP-13 gene expression in osteoblasts. The experimental groups were cultured with ginseng saponin in concentration of 1.0, 10, 25, 50, 100, 250 and $500{\mu}g/ml$ for MTT assay. Primary rat calvarial cells were pre-treated for 1 hour with ginseng saponin(100 ${\mu}g/ml$) and then stimulated with $IL-1{\beta}(1.0ng/ml)$ and PTH(10 nM). MMP-13 gene expression was evaluated by RT-PCR. The results were as follows: Ginseng saponin was cytotoxic to osteoblast at concentration exceeding $250{\mu}g/ml$ for longer than 24 hours in tissue culture(p<0.01). In RT-PCR analysis, steady state MMP-13 mRNA levels were increased approximately 350% by $IL-1{\beta}$, and 400% by PTH when normalized to untreated control. $IL-1{\beta}-indued$ MMP-13 mRNA expression was reduced 50% by pretreatment with ginseng saponin. But ginseng saponin didn't inhibit MMP-13 expression from PTH stimulated cells. This results suggest that ginseng saponin Inhibit $IL-1{\beta}-indued$ MMP-13 mRNA expression.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제21권1호
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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.

• 대한한방부인과학회지
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• 제29권2호
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• pp.1-14
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• 2016

Objectives : In this study, the author aimed to evaluate the effects of water extract of Dokwhalgisaeng-tang Gamibang (DGG) on osteoblast proliferation in murine calvarial cells. Methods : The osteoblast separated from murine calvariae was cultivated and evaluated the cell function. After the addition of DGG on the culture medium, we determined the effect of DGG on the cell proliferation, protein synthesis, alkaline phosphatase activity, collagen synthesis, and cell viability of the cultivated osteoblast in the presence of dexamethasone. Results : DGG increased the survival rate, proliferation, alkaline phosphatase (ALP) activity, protein synthesis and collagen synthesis of rat calvarial osteoblast in the presence of dexamethasone. Conclusions : DGG might improve the osteoporosis resulted from augmentation of osteoblast proliferation.

• Journal of Periodontal and Implant Science
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• 제28권4호
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• pp.769-786
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• 1998

The purpose of the present study is to examine the effect of cell proliferation and alkaline phosphatase activity in osteoblastic cells and to compare the bone healing ability of rat calvarial defects between the control group and the safflower seed extract treated group. Osteoblastic cells were obtained from calvariae of a fetal rat. Cells were cultured containing DMEM and safflower seed extract ($10^{-6}g/ml$, $10^{-3}g/ml$) at $37^{\circ}$ with 5% $CO_2$ in 100% humidity for 3 days. MTT was performed to examine the viability of the cells, and alkaline phosphatase activity was analyzed to examine the mineralization in vitro. Rat calvarial defects($5{\times}5mm$) in 250g Sprague-Dawly were made using round bur. Rats were administrated with safflower seed extract(0.35g/kg/day) for experimental periods. Calvarial defects were studied histopathologically and immunohistochemically at 1,4, and 8 weeks. High concentration group($10^{-3}g/ml$) of safflower seed extract significantly increased in the cell proliferation and alkaline phos phatase synthesis in osteoblastic cells. The infiltration of inflammatory cells and osteoclastic activities were decreased in the safflower seed extract treated group as compared with control group. Bone maturation was accelerated in the safflower seed extract treated group as compared to control group. No difference in osteoinductive process was observed between the control and the safflower seed extract treated group. Immunohistochemical observation revealed that protein expression of TGF-$\beta$and osteonectin during early healing phase in the safflower seed extract treated group was slightly increased as compared to control group. These results indicate that safflower seed extract promotes the healing process in bony defect of rat calvariae, and retains a potential applicability as an adjuvant therapeutic modality for regeneration of periodontal bony defect.

• International Journal of Oral Biology
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• 제37권1호
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• pp.9-16
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• 2012

The aim of this study was to evaluate surface characteristics and biological properties of the dentin -derived hydroxyapatite (HA) coating on titanium substrate. Dentinderived HA was obtained from extracted human teeth using a calcination method at $850^{\circ}C$. The commercially pure titanium (cp-Ti, ASTM Grade II) was used as a metallic substrate and a radio frequency magnetron sputtering method was employed as a coating method. Scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX) were utilized to investigate the coating aspects and composition. Atomic forced microscopy (AFM) and a surface profiler were used to assess the surface morphology and roughness. Corrosion tests were performed in phosphate-buffered saline at a $36.5{\pm}1^{\circ}C$ in order to determine the corrosion behavior of the uncoated and coated specimens. The biocompatibility of dentin-derived HA coated specimens with fetal rat calvarial cells and human gingival fibroblasts was assessed by SEM and cell proliferation analysis. The results showed that the dentin-derived HA coatings appeared to cover thinly and homogeneously the surfaces without changing of the titanium substrate. The EDX analysis of this the coating surface indicated the presence of Ca and P elements. The mean surface roughness of cp-Ti and dentin-derived coating specimens was $0.27{\mu}m$ and, $1.7{\mu}m$, respectively. Corrosion tests indicated a stable passive film of the dentin-derived HA coating specimens. SEM observations of fetal rat calvarial cells and human fibroblast cells on coated surfaces showed that the cells proliferated and developed a network of dense interconnections. The cells on all specimens proliferated actively within the culture period, showing good cell viability. At day 1 and 3, dentin-derived coating specimens showed 89% and 93% cell viability, respectively, when normalized to cp-Ti specimens. These results suggest that dentin-derived HA coating using the RF magnetron sputtering method has good surface characteristics and biocompatibility.

• 대한치과교정학회지
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• 제20권3호
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• pp.499-517
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• 1990

Electrical stimulation among several factors that influence bone remodeling has been studied by many investigators with great enthusiasm in orthodontic field. The action mechanisms of Pulsed Electromagnetic Field (PEMF) are different from those of the conventional electrode application method in that PEMF induces endogenous current in the living tissues. PEMF is known to have the healing effect in nonunion of bone and osteoporosis. It is widely used in orthopaedic scopes and the possibility of using the method in clinical orthodontics Is also conceivable. But the exact mechanisms by which the PEMF exerts its effects are not clearly understood. Therefore, the author wanted to see the effect of PEMF on five groups of rat calvarial cells obtained by sequential enzyme digestion method, and observed the changes in enzyme activation, collagen synthesis and $^3H-thymidine$ incorporation. The results were as follows: 1. Under the effect of PEMF, there were no changes in the alkaline phosphatase activity in five groups of cell populations. 2. Both the PEMF group and the PTH with PEMF group shelved no changes in acid phosphatase activities and there were no differences between two experimental groups. 3. Under the effect of PEMF, there was significant increase of collagen synthesis in the group V cell population. 4. Under the effect of PEMF, there were significant increases of $^3H-thymidine$ incorporation in the group IV and V cell populations.

• The Journal of Advanced Prosthodontics
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• 제7권6호
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• pp.484-495
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• 2015

PURPOSE. This study was to evaluate the effects of bacterial cellulose (BC) membranes as a barrier membrane on guided bone regeneration (GBR) in comparison with those of the resorbable collagen membranes. MATERIALS AND METHODS. BC membranes were fabricated using biomimetic technology. Surface properties were analyzed, Mechanical properties were measured, in vitro cell proliferation test were performed with NIH3T3 cells and in vivo study were performed with rat calvarial defect and histomorphometric analysis was done. The Mann-Whitney U test and the Wilcoxon signed rank test was used (${\alpha}<.05$). RESULTS. BC membrane showed significantly higher mechanical properties such as wet tensile strength than collagen membrane and represented a three-dimensional multilayered structure cross-linked by nano-fibers with 60 % porosity. In vitro study, cell adhesion and proliferation were observed on BC membrane. However, morphology of the cells was found to be less differentiated, and the cell proliferation rate was lower than those of the cells on collagen membrane. In vivo study, the grafted BC membrane did not induce inflammatory response, and maintained adequate space for bone regeneration. An amount of new bone formation in defect region loaded with BC membrane was significantly similar to that of collagen membrane application. CONCLUSION. BC membrane has potential to be used as a barrier membrane, and efficacy of the membrane on GBR is comparable to that of collagen membrane.