• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.28 no.3
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• pp.216-225
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• 2002

Hyaluronic acid (HA) is an almost essential component of extracellular matrices. Early in embryogenesis mesenchymal cells migrate, proliferate and differentiate, in part, because of the influence of HA. Since the features of embryogenesis are revisited during wound repair, including bone fracture repair, this study was initiated to evaluate whether HA has an effect on calcification and bone formation in an in vitro system of osteogenesis. Mouse calvaria Pre-osteoblast (MC3T3-E1) cells were cultured in ${\alpha}-MEM$ medium with microorganism-derivative hyaluronic acid that was produced by Strep. zooepidemicus which of molecular weight was 3 million units. The dosages were categorized in each 0.5, 1.0 and 2.0 mg/ml concentration experimental groups. After 2 and 4 days cultures in expeirmental and control groups, the tendency of cell proliferation, MTT assay, protein synthesis ability, collagen synthesis and alkaline phosphatase activity were analysed and bone nodule formation capacity were measured with Alizarin Red S stain after 29 days cultures. The cell proliferation was increased in time, especially the group of 0.5 and 1.0 mg/ml concentration of HA were showed prominent cell proliferation. After 2 and 4 days culture, experimental groups in general were greater cell activity in MTT assay. The protein synthesis was increased in all experimental groups compared to control group, especially most prominent in 1.0 mg/ml concentration group. The collagen synthesis capacity were increased in HA experimental groups, especially prominent in 1.0 mg/ml group and the activity of alkaline phosphatase were increased, especially also prominent in 1.0 mg/ml group, compared to control group. Above these, the activity of mouse carvarial pre-osteoblast cells was showed greater bone osteogenesis activity in all applied HA experimental group, especially group of 1.0 mg/ml concentration of HA.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.40
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• pp.34.1-34.8
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• 2018

Background: Radiation therapy is widely employed in the treatment of head and neck cancer. Adverse effects of therapeutic irradiation include delayed bone healing after dental extraction or impaired bone regeneration at the irradiated bony defect. Development of a reliable experimental model may be beneficial to study tissue regeneration in the irradiated field. The current study aimed to develop a relevant animal model of post-radiation cranial bone defect. Methods: A lead shielding block was designed for selective external irradiation of the mouse calvaria. Critical-size calvarial defect was created 2 weeks after the irradiation. The defect was filled with a collagen scaffold, with or without incorporation of bone morphogenetic protein 2 (BMP-2) (1 ㎍/ml). The non-irradiated mice treated with or without BMP-2-included scaffold served as control. Four weeks after the surgery, the specimens were harvested and the degree of bone formation was evaluated by histological and radiographical examinations. Results: BMP-2-treated scaffold yielded significant bone regeneration in the mice calvarial defects. However, a single fraction of external irradiation was observed to eliminate the bone regeneration capacity of the BMP-2-incorporated scaffold without influencing the survival of the animals. Conclusion: The current study established an efficient model for post-radiation cranial bone regeneration and can be applied for evaluating the robust bone formation system using various chemokines or agents in unfavorable, demanding radiation-related bone defect models.

• Journal of Periodontal and Implant Science
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• v.35 no.2
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• pp.491-510
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• 2005

This study was performed to evaluate the effect of inorganic polyphosphate on bone formation in the calvaria of rabbit in the procedure of guided bone regeneration with bovine cancellous bone graft and titanium reinforced expanded polytetrafluoroethylene(TR-ePTFE) membrane. The rabbits were divided into four groups. Control group I used only TR-ePTFE membrane, control group II used TR-ePTFE membrane and deproteinized bovine bone mineral soaked in saline, experimental group III and IV used TR-ePTFE membrane and deproteinized bovine bone mineral soaked in 1% or 2% inorganic polyphosphate respectively. After decortication in the calvaria, GBR procedure was performed on 12 rabbits with titanium reinforced ePTFE membrane filled with deproteinized bovine bone mineral soaked in saline or inorganic polyphosphate. The animals were sacrificed at 2 weeks, 4 weeks, and 8 weeks after the surgery. Decalcified and non-decalcified specimens were processed for histologic and immunohistochemistric analysis. 1. Titanium reinforced ePTFE(TR-ePTFE) membrane showed good spacemaking and cell occlusiveness capability, but it showed poor wound stabilization. 2. The deproteinized bovine bone mineral did not promote bone regeneration, but it acted as a space filler. 3. There was no complete resorption of the deproteinized bovine bone mineral within 8 weeks. 4. 1% inorganic polyphosphate did not promote bone formation, but 2% inorganic polyphosphate promoted bone formation. Within the above results, 2% inorganic polyphosphate could be used effectively for bone regeneration.

• 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 Veterinary Clinics
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• v.17 no.2
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• pp.470-474
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• 2000

A Shih Tzu Puppy had clinical onset of anotexia crying and progression of neurological sings when enlargement of the cranial vault at 1 month old and died after showing clinical signs during 2 months period. Radiological and pathological examinations were performed. Radiological findings were homogeneous appearance of the calvaria with cortical thinning, loss of the normal convolutional skull markings and persistent fontanelles. Grossly enlargement of the cranial vault thinning of the bone and defective closure of the fontanelles were also observed. The entire subcortical area of the cerebral hemispheres with severe, dilatation of ventricles and cerebrospinal fluid(CSF) wits absent. There was parenchyma atrophy affecting chiefly in the white mater and the cerebral cortices, axon degeneration and necrosis and gitter cell infiltration in the whiter matter and the subependymal area. Mononuclear perivacular cuffing in the cerebrum and the pons was shown. Based on the radiological, gross and histopathological findings, this case was believed to have congenital hydrocephalus with nonsuppurative encephahitis. Possible etiology on the case is also discussed.

• The Korean Journal of Pharmacology
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• v.26 no.2
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• pp.193-207
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• 1990

Transforming growth factor ${\beta}(TGF-{\beta})$ is a multifunctional polypeptide with diverse effects on the proliferation, differentiation and other functions in many cell types. $TGF-{\beta}$ is highly abundant in bone matrix and induces divergent responses in many aspects of bone cell metabolism . Several lines of investigation indicate that matrix-associated $TGF-{\beta}$ is the products of bone cells themselves. However, exact bone cell type reponsible for the production of $TGF-{\beta}$ is still in controversy, The present study was undertaken to determine the cellular origin of matrix-associated $TGF-{\beta}$ and to assess how different bone cells respond to $TGF-{\beta}$. As a prerequisite for this, 5 bone cell populations of distinct phenotype were isolated from fetal calvaria with sequential enzyme digestion protocol and biochemical characterization. Calvarial cell populations released in early stage showed fibroblastic features whereas populations relesed later was enriched with osteoblast-like cell as judged by their acid and alkaline phosphatase activities, cAMP responsiveness to parathyroid hormone, calcitonin and prostaglandin $E_2$ and collagen synthesis rate. By polyacylamide gel and immunoblot analysis of bone and calvarial cell extracts, presence of $TGF-{\beta}$ in bone tissues and production of $TGF-{\beta}$ by bone cells were confirmed again. Subsequent analysis of calvarial cell extracts prepared as individual population revealed that all calvarial cell populations synthesize $TGF-{\beta}$. Exogenously added $TGF-{\beta}$ induced biphasic response upon bone cell proliferation under serum-free condition. In osteoblastic cell populations, it was stimulatory whereas inhibitory in fibroblastic cell populations. In contrast, collagen and noncollagen protein synthesis of all calvarial cell populations were stimulated by $TGF-{\beta}$. Enhancement of protein synthesis was found to be more general rather than specific for collagen synthesis. In addition, effects of $TGF-{\beta}$ on protein synthesis were independent to its effects on cell proliferation. In summary, production of $TGF-{\beta}$ by bone cells and differential actions on various cell populations observed in this study suggest that $TGF-{\beta}$ may play an important role in the regulation of bone metabolism by modulating the specific cellular functions in autocrine and paracrine fashion.

• Progress in Medical Physics
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• v.21 no.2
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• pp.137-144
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• 2010

The purpose of this study was to evaluate the usefulness of reducing of craniofacial radiation dose using automatic exposure control (AEC) technique in the 64 multi-detector computed tomography (MDCT). We used SOMATOM Definition 64 multi-detector CT, and head of whole body phantom (KUPBU-50, Kyoto Kagaku CO. Ltd). The protocol were helical scan method with 120 kVp, 1 sec of rotation time, 5 mm of slice thickness and increment, 250 mm of FOV, $512{\times}512$ of matrix size, $64{\times}0.625\;mm$ of collimation, and 1 of pitch. The evaluation of dose reducing effect was compared the fixed tube current of 350 with AEC technique. The image quality was measured the noise using standard deviation of CT number. The range of craniofacial bone was to mentum end from calvaria apex, which devided three regions: calvaria~superciliary ridge (1 segment), superciliary ridge~acanthion (2 segment), and acanthion~mentum (3 segment). In the fixed tube current technique, CTDIvol was 57.7 mGy, DLP was $640.2\;mGy{\cdot}cm$ in the all regions. The AEC technique was showed that 1 segment were 30.7 mGy of CTDIvol, 340.7 $mGy{\cdot}cm$ of DLP, 2 segment were 46.5 mGy of CTDIvol, $515.0\;mGy{\cdot}cm$ of DLP, and 3 segment were 30.3 mGy of CTDIvol, $337.0\;mGy{\cdot}cm$ of DLP. The standard deviation of CT number was 2.622 with the fixed tube current technique and 3.023 with the AEC technique in the 1 segment, was 3.118 with the fixed tube current technique and 3.379 with the AEC technique in the 2 segment, was 2.670 with the fixed tube current technique and 3.186 with the AEC technique in the 3 segment. The craniofacial radiation dose using AEC Technique in the 64 MDCT was evaluated the usefulness of reducing for the eye, the parotid and thyroid with high radiation sensitivity particularly.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.4
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• pp.313-323
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• 2010

Purpose: This study was conducted to evaluate the effect of beta-tricalcium phosphate (Cerasorb$^{(R)}$, Germany) and deproteinized bovine bone (Bio-Oss$^{(R)}$, Switzerland) grafted to the defect of rat calvaria artificially created and the effect of use of absorbable membrane (BioMesh$^{(R)}$, Korea) on new bone formation. Materials and Methods: Transosseous circular calvarial defects with diameters of 5 mm were prepared in the both parietal bone of 30 rats. In the control group I, no specific treatment was done on the defects. In the control group II, the defects were covered with absorbable membrane. In the experimental group I, deproteinized bovine bone was grafted without absorbable membrane; in the experimental group II, deproteinized bovine bone was grafted with absorbable membrane; in the experimental group III, beta-tricalcium phosphate was grafted without absorbable membrane; in the experimental group IV, beta-tricalcium phosphate was grafted with absorbable membrane. The animals were sacrificed after 3 weeks and 6 weeks respectively, and histologic and histomorphometric evaluations were performed. Results: Compare to the control groups, the experimental groups showed more newly formed bone. Between the experimental groups, beta-tricalcium phosphate showed more resorption than deproteinized bovine bone. Stabilization of grafted material and interception of the soft tissue invasion was observed in the specimen treated with membrane. There was no statistical difference between the experimental group I, III and experimental group II, IV classified by graft material, but statistically significant increase in the amount of newly formed bone was observed in the experimental group I, II and II, IV classified by the use of membrane (P<0.05). Conclusion: Both beta-tricalcium phosphate and deproteinized bovine bone showed similar osteoconductibility, but beta-tricalcium phosphate is thought to be closer to ideal synthetic graft material because it showed higher resorption rate in vivo. Increased new bone formation can be expected in bone graft with use of membrane.

• Journal of Periodontal and Implant Science
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• v.37 no.1
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• pp.77-89
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• 2007

This study was performed to evaluate the effect of deproteinized bovine bone mineral soaked in inorganic polyphosphate on bone regeneration in the calvaria of rabbit in the procedure of guided bone regeneration with titanium reinforced expanded polytetrafluoroethylene(TR-ePTFE) membrane. The rabbits were divided into four groups. Control group used TR-ePTFE membrane filled with de-proteinized bovine bone mineral, experimental group I used TR-ePTFE membrane and deproteinized bovine bone mineral soaked in 4% inorganic polyphosphate, experimental group II and III used TR-ePTFE membrane and deproteinized bovine bone mineral soaked in 8% or 16% inorganic poly-phosphate respectively. After decortication in the calvaria, GBR procedure was performed on 8 rabbits with only TR-ePTFE membrane or titanium reinforced ePTFE membrane filled with deproteinized bovine bone mineral soaked in inorganic polyphosphate. The animals were sacrificed at 4 weeks, and 8 weeks af-ter the surgery. Non-decalcified specimens were processed for histologic analysis, and new bone for-mation was assessed by histomorphometric as well as statical analysis. 1. Both control group and experirrental group dermnstrated increasing of new bone formation until 8weeks. 2. At 8 weeks, experimental group I and group II showed the significant difference compared to control group in new bone formation. Especially experimental group II showed the most in-creasing of new bone formation. 3. The higher concentration of inorganic polyphosphate filled, the more volume of bone formation pro-moted, but experimental group III did not reveal significant difference compared to contol group. 4. Deproteinized bovine bone mineral did not resorbed at all until 8 weeks. These results suggest that inorganic polyphosphate has a promoting effect on bone regeneration. possibly by enhancing osteoconductivity of the carrier and by increasing osteoinductivity of the defected alveolar bone tissue, but not as we respect.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.34 no.6
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• pp.391-397
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• 2012

Purpose: The purpose of this study was to evaluate the effectiveness of the platelet-rich fibrin (PRF) used in combination with the porcine cancellous bone as a scaffold, in promoting bone regeneration in the bone defects ofthe rabbit calvaria. Methods: Ten rabbits were used in the study. Three round-shaped defects (diameter 8.0 mm) were created in the rabbit calvaria and were filled with nothing (control group), porcine cancellousbone (Experimental Group 1, porcine bone) and PRF-mixed porcine cancellous bone (Experimental Group 2). TS-GBB is a xenogenic bone-substitute product comprised of a high heat-treated mineralized porcine cancellous bone. Animals were sacrificed at 6 weeks and 12 weeks for the histological and radiographic evaluations. Results: In the micro computed tomography and histological results, the experimental groups 1 and 2 showed more bone formation, remodeling, and calcification than the control group. The new bone formation ratio showed theGroup 2 to be larger than Group 1 at6 and 12 weeks. However, there was no significant difference between the experimental groups 1 and 2 in the new bone formation area, at the 6 and 12 weeks (P>0.05). Conclusion: The PRF-mixed group showed more bone formation than the porcine cancellousbonegroup (TS-GBB), butthere was a no significant difference. The PRF may not lead to enhanced bone healing when grafted with the porcine cancellous bone.