• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.33 no.5
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• pp.470-478
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• 2007

The purpose of this study was to investigate the clinical, biomechanical, and histologic changes in new distraction osteogenesis(DO) technique combined with a compression stimulation in accordance to different compression-distraction force ratio. 23 adult male rabbits underwent open-osteotomy at the mandibular body area and a external distraction device was applied. In the control group of 8 rabbits, only a 8 mm of distraction was performed by conventional DO technique. In an experimental group of 15 rabbits, a distraction followed by a compression force was performed according to the ratio of compression-distraction suggested by authors. The rate of experimental group I was set up as a 2 mm compression versus 10 mm distraction and the rate of experimental group II was set up as a 3 mm compression versus 11 mm distraction. All the rabbits were sacrificed for a gross finding, biomechanical, histomorphometric and histologic findings at the time of 55 days from the operation day. The results were as follows: 1. On the gross findings, because all rabbits had a sufficient healing time, every distracted new bone had good bone quality and we could not find any difference among all three groups. 2. In the histologic findings, rapid bone maturation(wide lamellar bone formation in the cancellous and cortical bone areas) was observed in two experimental groups compared to the control group. 3. On the bone density tests, the experimental group II showed higher bone density than the other experimental group and control group(control group-$0,2906g/cm^2$, experimental group I-$0.2961g/cm^2$, experimental group II-$0.3328g/cm^2$). 4. On the biomechanical tests, the experimental group II had significantly higher bone microhardness than the other experimental group and control group(control group-252.7 MPa, experimental group I-263.5 MPa, experimental group II-426.0 MPa). 5. On the microhardness tests, when we compared the hardness ratio of distracted bone versus normal bone, we could find experimental group II had significantly higher hardness ratio than the other experimental group and control group(control group-0.47, experimental group I-0.575, experimental group II-0.80). From this study, we could deduce that the modified distraction osteogenesis method with a compression stimulation might improve the quality of bone regeneration and shorten the consolidation period in comparison with conventional distraction osteogenesis techniques.

• The Journal of Advanced Prosthodontics
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• v.6 no.2
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• pp.96-102
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• 2014

PURPOSE. This study was performed to characterize the effects of zirconia coated with calcium phosphate and hydroxyapatite compared to smooth zirconia after bone marrow-derived osteoblast culture. MATERIALS AND METHODS. Bone marrow-derived osteoblasts were cultured on (1) smooth zirconia, (2) zirconia coated with calcium phosphate (CaP), and (3) zirconia coated with hydroxyapatite (HA). The tetrazolium-based colorimetric assay (MTT test) was used for cell proliferation evaluation. Scanning electron microscopy (SEM) and alkaline phosphatase (ALP) activity was measured to evaluate the cellular morphology and differentiation rate. X-ray photoelectron spectroscopy (XPS) was employed for the analysis of surface chemistry. The genetic expression of the osteoblasts and dissolution behavior of the coatings were observed. Assessment of the significance level of the differences between the groups was done with analysis of variance (ANOVA). RESULTS. From the MTT assay, no significant difference between smooth and surface coated zirconia was found (P>.05). From the SEM image, cells on all three groups of discs were sporadically triangular or spread out in shape with formation of filopodia. From the ALP activity assay, the optical density of osteoblasts on smooth zirconia discs was higher than that on surface treated zirconia discs (P>.05). Most of the genes related to cell adhesion showed similar expression level between smooth and surface treated zirconia. The dissolution rate was higher with CaP than HA coating. CONCLUSION. The attachment and growth behavior of bone-marrow-derived osteoblasts cultured on smooth surface coated zirconia showed comparable results. However, the HA coating showed more time-dependent stability compared to the CaP coating.

• Journal of Biomedical Engineering Research
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• v.25 no.3
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• pp.201-206
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• 2004

The bone formation and re-formation are regulated by two factors that are the synthesis of bone matrix by osteoblast and reabsorption by osteoclast. Recently, there are many studies about regeneration and healing of bone tissue by activation of osteoblast. In general, it is known that the activation of osteoblast is influenced by not only biological stimulus but physical stimulus. In this study, we verified that ostoeblast activation was influenced by low intensity ultrasound. Various ultrasonic properties were used to find out the most appropriate condition on cell activation. From this study, we could confirm that 0.3W/$\textrm{cm}^2$ intensity of ultrasound was the most appropriate to tell activation over whole duty cycles and the increasing rate of tell was the highest at 50％ duty cycle. Thus, it is expected that optimal ultrasonic characteristics on regeneration of bone matrix may be applied to fracture and osteoporosis healing.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.1
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• pp.13-23
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• 2010

Objective: The present study was carried out to induce differentiation of human embryonic stem cells (hESCs) into germ cells and to establish a culture condition for single hESCs dissociated by enzyme. Methods: Embryonic body (EB) was formed by hanging drop culture for 3 days from hESCs colony. The EBs were cultured in the medium supplemented with retionic acid (RA) or/and bone morphogenetic protein-4 (BMP4) for 14 days to differentiate into germ cells. Germ cell specific markers, c-kit and VASA were used for immunohistochemistry of EB. Human ESCs colonies were dissociated into single cells by Collagenase, Tryple and Accutase, and then colony formation rate of the single cells was examined. Rho-associated kinase inhibitor (ROCK inhibitor, Y27632) was added into the culture medium of single cells to reduce the apoptotic damage during the dissociation. Results: Single cells dissociated with Tryple or Accutase showed higher colony formation rates compared to the cells dissociated with Collagenase. Seeding of $5{\times}10^3$ cells/well (4 well dish) was efficient to obtain high colony formation rate compared to other concentrations of seeding cell. Addition of Y27632 significantly increased the colony formation rate of the single cells dissociated by Tryple. Immunohistochemistry of EB with c-kit and VASA markers showed a weak fluorescence signals compared to the signals from the testicular tissue. Conclusion: Dissociation with Tryple was useful to obtain healthy single cells and addition of Y27632 was beneficial for survival and colony formation of the single cells. Unlike other studies, we just observed a dim fluorescence staining of the germ cell markers, probably caused by the short-term culture for the differentiation of EB compared to other studies.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.4
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• pp.306-312
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• 2010

Purpose: The purpose of this experiment was to evaluate the clinical effect of cultured autoglogous osteoblasts as a way to treat the defect of mandible in rabbits. Materials and Methods: Twelve rabbits were used to determine the rate of osteogenesis. The osteoblasts were obtained from the iliac crest of rabbits using aspiration. They were then cultured in Dulbecco's Modified Eagles's Medium (DMEM) with beta-glycerophosophatate, L-ascorbicacid, and dexamethasone to proliferate and differentiate osteoprogenitor cells. The expression of osteogenic markers were detected by reverse transcription-polymerase chain reaction (RT-PCR) and silver nitrate staining techniques. Five, 10-mm holes were placed in each rabbit mandible to simulate defective regions with the use of a low speed trephine bur. In the experimental group, the previously cited defects were grafted with both activated osteoblastic and autogenous bone. The control group, however, was only grafted with autogenous bone. Both groups were then analyzed at 2, 4, and 8-week intervals using bone histomorphometric analysis. Results: According to histomorphologic analysis, the rates of new bone formation at the 2, 4, and 8-week intervals were 36%, 51%, and 23% for the control group, respectively; 52%, 39%, and 28%, for the experimental group, respectively. The experimental group showed higher rates of new bone formation compared to the control group at both the 2-week and 8-week interval. Conclusion: Bone marrow-derived osteoblasts seems to be a promising bone graft material.

• BMB Reports
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• v.53 no.3
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• pp.154-159
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• 2020

We investigated the effects of physalin A, B, D, and F on osteoclastogenesis induced by receptor activator of nuclear factor κB ligand (RANKL). The biological functions of different physalins were first predicted using an in silico bioinformatic tool (BATMAN-TCM). Afterwards, we tested cell viability and cell apoptosis rate to analyze the cytotoxicity of different physalins. We analyzed the inhibitory effects of physalins on RANKL-induced osteoclastogenesis from mouse bone-marrow macrophages (BMMs) using a tartrate-resistant acid phosphatase (TRAP) stain. We found that physalin D has the best selectivity index (SI) among all analyzed physalins. We then confirmed the inhibitory effects of physalin D on osteoclast maturation and function by immunostaining of F-actin and a pit-formation assay. On the molecular level, physalin D attenuated RANKL-evoked intracellular calcium ([Ca(2+)](i)) oscillation by inhibiting phosphorylation of phospholipase Cγ2 (PLCγ2) and thus blocked the downstream activation of Ca2+/calmodulin-dependent protein kinases (CaMK)IV and cAMP-responsive element-binding protein (CREB). An animal study showed that physalin D treatment rescues bone microarchitecture, prevents bone loss, and restores bone strength in a model of rapid bone loss induced by soluble RANKL. Taken together, these results suggest that physalin D inhibits RANKL-induced osteoclastogenesis and bone loss via suppressing the PLCγ2-CaMK-CREB pathway.

• The Journal of Korean Academy of Prosthodontics
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• v.51 no.3
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• pp.175-182
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• 2013

Purpose: The purpose of this study was to investigate the effect of different administration duration of alendronate on initial wound healing and new bone formation of extraction socket in rats. Materials and methods: Fifteen male Sprague-Dawley rats (body weight 130-140 g, 4 weeks old, male) were divided into control group (no alendronate administration) and experimental group (alendronate administration). Experimental group was subdivided into 1 week administrated group, 2 week administrated group, 4 week administrated group and 6 week administrated group according to duration of administration. For the experimental groups, during the designated time period (at the time of extraction, 1 week before extraction, 3 week before extraction and 5 week before extraction) till 1 week after extraction, rats were subcutaneously injected with Alendronate at the dose of 1.0 mg/Kg three times a week. Each specimen from 6 week experimental group and control group were used for microarray analysis, and other specimens were used for histological analysis. The rate of new bone formation within the extraction site and bone loss activity was analyzed using TRAP staining. Statistical analysis was performed using Kruskal Wallis test. (${\alpha}=.05$) Results: After one week from the time of extraction, the rate of new bone formation within extraction site for the control group ($16.77%{\pm}1.36%$) compared to the 4 week experimental group ($14.99%{\pm}6.26%$) was lower. However, no statistically significant difference was found. Increase in the number of inactive lacuna (empty lacuna) and decrease in the number of TRAP positive cell were identified with increased duration of administration. There was no significant difference. Conclusion: The results of this study showed as the duration of Alendronate administration increased the rate of new bone formation decreased with loss of bone activity and reduced number of osteoclast.

• Journal of the Korean Academy of Esthetic Dentistry
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• v.8 no.1
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• pp.36-44
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• 1999

No one really doubts that the hottest area of dentistry in 21st century would most likely be 'implant'. With the support of a vast amount of research, implant has been successfully and rapidly absorbed into the field of general dentistry including private practitioners. For teeth with hopeless(or refractory) periodontitis or periapical pathosis, with no hesitation most dentists would think extraction as the sole treatment option followed by prosthodontic replacement possibly including implant. Not many dentists would take Trasplantation/Replantation as another treatment option for a particular condition. Dentistry is often more focused on 'Restoration' than 'Preservation' of natural dentition. 'Biologic Implant' is obviously much closer to the concept of 'Preservation'. Many different types of biologic implant system have been introduced to clinical dentistry so far. Many of those have failed to earn reasonable acknowledgement despite of the clinical success they brought. For some reason biologic implant has rather been alienated for long time. RPI(Ring Pin Implant) is designed to improve the prognosis and success rate of transplanted/replanted teeth. RPI is a Ti-based custom made implant system. It is fabricated either by electric casting or milling process. The major feature RPI gas is the 'ring & hole' structure. The hole should be no less than 1mm diameter to allow bone bridge formation thru it. The ring structure and bone bridge formation creates anti-torque activity, which largely increases the 'initial stability' of the transplanted/replanted teeth. It is also reported that RPI is beneficial in the aspect of resisting root resorption following replantation/transplantation procedure.

• The Journal of Korean Obstetrics and Gynecology
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• v.27 no.2
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• pp.23-33
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• 2014

Objectives: In this study, the author aimed to evaluate the effect of EtOH extract of Ganoderma lucidum (GLE) on osteoblast proliferation in rat fetus calvarial cells. Methods: The osteoblast separated from rat fetus calvariae was cultivated for 6~21 days and evaluated the cell function. After the addition of GLE on the culture medium, we determined the effect of GLE on the cell viability, cell proliferation, bone matrix protein synthesis, alkaline phosphatase (ALP) activity, collagen synthesis and calcified nodule formation of the cultivated osteoblast. Results: GLE did not change the survival rate of rat calvarial osteoblast. GLE increased the proliferation of rat calvarial osteoblast. GLE increased ALP activity of rat calvarial osteoblast. GLE increased bone matrix protein synthesis of rat calvarial osteoblast. GLE increased collagen synthesis of rat calvarial osteoblast. GLE slightly affected calcified nodule formation of rat calvarial osteoblast. Conclusions: This study suggests that Ganoderma lucidum might improve the osteoporosis resulted from augmentation of osteoblast proliferation.

• Journal of Periodontal and Implant Science
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• v.29 no.3
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• pp.483-509
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• 1999

Biodegradable barrier membrane has been demonstrated to have guided bone regeneration capacity on the animal study. The purpose of this study is to evaluate the effects of cultured calvarial cell inoculated on the biodegradable barrier membrane for the regeneration of the artificial bone defect. In this experiment 35 Sprague-Dawley male rats(mean BW 150gm) were used. 30 rats were divided into 3 groups. In group I, defects were covered periosteum without membrane. In group II, defects were repaired using biodegradable barrier membrane. In group III, the defects were repaired using biodegradable barrier membrane seeded with cultured calvarial cell. Every surgical procedure were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). After anesthesia, 5 rats were sacrificed by decapitation 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. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. The membrane were inserted on the artificial bone defect after 3 days of culture. A single 3-mm diameter full-thickness artificial calvarial defect was made in each animal by using with bone trephine drill. After the every surgical intervention of animal, all of the animals were sacrificed at 1, 2, 3 weeks after surgery by using of perfusion technique. For obtaining histological section, tissues were fixed in 2.5% Glutaraldehyde (0.1M cacodylate buffer, pH 7.2) and Karnovsky's fixative solution, and decalcified with 0.1M disodium ethylene diaminetetraacetate for 3 weeks. Tissue embeding was performed in paraffin and cut parallel to the surface of calvaria. Section in 7${\mu}m$ thickness of tissue was done and stained with Hematoxylin-Eosin. All the specimens were observed under the light microscopy. The following results were obtained. 1 . During the whole period of experiment, fibrous connective tissue was revealed at 1week after surgery which meant rapid soft tissue recovery. The healing rate of defected area into new bone formation of the test group was observed more rapid tendency than other two groups. 2 . The sequence of healing rate of bone defected area was as follows ； test group, positive control, negative control group. 3 . During the experiment, an osteoclastic cell around preexisted bone was not found. New bone formation was originated from the periphery of the remaing bone wall, and gradually extended into central portion of the bone defect. 4 . The biodegradable barrier membrane was observed favorable biocompatibility during this experimental period without any other noticeable foreign body reaction. And mineralization in the newly formed osteoid tissue revealed relatively more rapid than other group since early stage of the healing process. Conclusively, the cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of regeneration of artificial bone defects of alveolar bone. This study thus demonstrates a tissue-engineering the approach to the repair of bone defects, which may have clinical applications in clinical fields of the dentistry including periodontics.