• 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.24 no.3
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• pp.671-689
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• 1994

The purpose of this study was to compare effects of various bone grafts on periodontal regeneration of alveolar bone defects in dogs. Seven adult dogs aged 12 to 18 months were used in this study. Experimental alveolar bone defects were created surgically with a #1/2 round bur at the furcation area of the buccal surface of the mandibular 3rd, 4th premolars and 1st molar. Each experimental alveolar bone defects were grafted with dense hydroxyapatite, natural coral, and decalcified freeze-dried bone, and respectively divided into DHA, NC, DFDB group. An area without bone graft was divided into control group. At 1,2,4,6, and 12 weeks, dogs were serially sacrificed and specimens were prepared with Hematoxylin-Eosin stain and Mallory stain for light microscopic evaluation. The results of this study were as follows : 1. In control group, the matrix change of granulation tissue was observed at 1 week. And in experimental groups, the appearance of connective tissue around graft materials was loosely formed at 1 week, but densely formed at 2 weeks. 2. In every group, the slight formation of new trabecular bone was seen from remaining bone at 1 week. 3. The DHA and NC particles were gradually encapsulated by new trabecular bone from remaining bone, and the osteoid tissue was directly induced from DFDB particles. 4. The presence of osteoblasts was first observed at 1 week in control group and at 2 weeks in NC group, but at 6 weeks in DHA group. 5. In DHA group, the resorption of particles was not observed until 12 weeks. But in NC and DFDB group, the particles were resorbed at 6 weeks and replaced by new bone. And the amount and size of particles were reduced, and their border represented irregular form. In summary, in three experimental groups the inflammatory or foreign body reaction were slight, but the regeneration of new osteoid tissue and the matrix change of dense connective tissue fiber were observed. Especially, NC and DFDB materials were considered as the biocompatible graft materials which were effective in the regenertion of new bone.

• Macromolecular Research
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• v.10 no.3
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• pp.158-167
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• 2002

In order to endow with new bioactive functionality from small intestine submucosa (SIS) powder as natural source to poly (L-lactide) (PLA) and poly (lactide-co-glycolide) (PLGA) synthetic biodegradable polymer, porous SIS/PLA and SIS/PLGA as natural/synthetic composite scaffolds were prepared by means of the solvent casting/salt leaching methods for the possibility of the application of tissue engineered bone and cartilage. A uniform distribution of good interconnected pores from the surface to core region was observed the pore size of 40~500 ${\mu}{\textrm}{m}$ independent with SIS amount using the solvent casting/salt leaching method. Porosities, specific pore areas as well as pore size distribution also were almost same. After the fabrication of SIS/PLA hybrid scaffolds, the wetting properties was greatly enhanced resulting in more uniform cell seeding and distribution. Five groups as PGA non-woven mesh without glutaraldehyde (GA) treatment, PLA scaffold without or with GA treatment, and SIS/PLA (Code No.3 ; 1 : 12 of salt content, (0.4 : 1 of SIS content, and 144 ${\mu}{\textrm}{m}$ of median pore size) without or with GA treatment were implanted into the back of nude mouse to observe the effect of SIS on the induction of cells proliferation by hematoxylin and eosin, and von Kossa staining for 8 weeks. It was observed that the effect of SIS/PLA scaffolds with GA treatment on bone induction are stronger than PLA scaffolds, that is to say, in the order of PLA/SIS scaffolds with GA treatment > PLA/SIS scaffolds without GA treatment > PGA nonwoven > PLA scaffolds only with GA treatment = PLA scaffolds only without GA treatment for the osteoinduction activity. The possible explanations are (1) many kinds of secreted, circulating, and extracellular matrix-bound growth factors from SIS to significantly affect critical processes of tissue development and differentiation, (2) the exposure of SIS to GA resulted in significantly calcification, and (3) peri-implant fibrosis due to covalent bonding between collagen molecule by crosslinking reaction. In conclusion, it seems that SIS plays an important role for bone induction in SIS/PLA scaffolds for the application of tissue engineering area.

• Proceedings of the Korean Society of Life Science Conference
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• 2003.10a
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• pp.28-37
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• 2003

Tissue engineering and stem cells show potentials to restore lost or malfunctioning human tissues or organs. Another cell source for tissue engineering of cardiovascular tissues is stem cell. This study reports the development of cardiovascular tissues using tissue engineering and mesenchymal stem cells. The blood vessels and heart valves were fabricated by culturing mesenchymal stem cells on biodegradable synthetic or natural matrices. Bone marrow was isolated from dogs or rats and mesenchymal stem cells were cultured. The cells were seeded onto biodegradable synthetic or natural matrices and implanted in dogs. Histological and immunohistochemical analyses were performed to examine the regenerated cardiovascular tissues. Histological and immunohistochemical analyses showed the complete regeneration of blood vessels and heart valves. Fluorescent labeling of cells prior to implantation and fluorescence examination of the regenerated tissues revealed that the implanted cells reconstituted the cardiovascular tissues. This study demonstrates the potential of tissue engineering and mesenchymal stem cells for the regeneration of functional cardiovascular tissues or organs.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.32 no.5
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• pp.478-483
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• 2010

Bisphosphonates are synthetic analogue and have high affinity on bone remodeling site. Since they have a long half-reduction time, they accumulate at bone and act for a long time. They are widely used in osteo-porosis derived from imflammatory bowel disease or postmenopausal osteoporosis patient for bone mineral density improvement. In addition, they neutralize hypercalcemia owing to bone metastasis of malignancy. However, a jaw bone necrosis was recently reported in some patients who have taken bisphosphonates for a long time. It is called Bisphosphonate Related Osteonecrosis of Jaws (BRONJ). It can come spontaneous-ly, but more often after oral surgery including tooth extraction. In this case, a 80-year-old woman was treated with bisphosphonate (sodium alendronate) for 2 years to improve bone mineral density. She had her left lower second molar tooth extracted at local clinic. After extraction, she had inflammatory symptoms like a pain, a cheek swelling, and a discharge of pus. She was referred to our clinic for treatment. We treated meticulously from dressing to surgery. After following up about 1.5 years, the jaw lesion was successfully healed. So we report this case.

• Journal of Periodontal and Implant Science
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• v.41 no.2
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• pp.67-72
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• 2011

Purpose: The aim of this study is to determine whether certain biomaterials have the potential to support cell attachment. After seeding bone marrow stromal cells onto the biomaterials, we investigated their responses to each material in vitro. Methods: Rat bone marrow derived stromal cells were used. The biomaterials were deproteinized bovine bone mineral (DBBM), DBBM coated with fibronectin (FN), synthetic hydroxyapatite (HA), HA coated with FN, HA coated with $\beta$-tricalcium phosphate (TCP), and pure $\beta$-TCP. With confocal laser scanning microscopy, actin filaments and vinculin were observed after 6, 12, and 24 hours of cell seeding. The morphological features of cells on each biomaterial were observed using scanning electron microscopy at day 1 and 7. Results: The cells on HA/FN and HA spread widely and showed better defined actin cytoskeletons than those on the other biomaterials. At the initial phase, FN seemed to have a favorable effect on cell adhesion. In DBBM, very few cells adhered to the surface. Conclusions: Within the limitations of this study, we can conclude that in contrast with DBBM not supporting cell attachment, HA provided a more favorable environment with respect to cell attachment.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.33 no.4
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• pp.308-313
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• 2011

Purpose: Beta-tricalcium phosphate (${\beta}$-TCP) is a synthetic calcium phosphate ceramic that has widely been used as a bone material to repair bone defects. Despite many clinical studies, the molecular mechanism whereby this biomaterial alters the gene expression in osteoblasts to promote bone formation is poorly understood. Thus, we attempted to address this question by using microarray techniques to identify the genes that are differentially regulated in osteoblasts exposed to ${\beta}$-TCP. Methods: By using DNA microarrays, we identified several genes whose expression levels were significantly up- or down-regulated in osteoblast-likeMG-63cells cultured with ${\beta}$-TCP at a concentration of 100 mg/10 ml for 24 hours. Results: The differentially expressed genes covered a broad range of functional activities: signal transduction, transcription, cell cycle regulation, vesicular transport, apoptosis, immunity, cytoskeletal elements and cell proliferation and differentiation. Conclusion: The gene expression changes related to cell proliferation and differentiation, vesicle transport, immunity and defense could affect the osteogenic activities of osteoblasts for bone regeneration. However, further studies will be required to verify the relative importance of these genes in bone formation, their temporal and spatial expression patterns and their interactions with each other.

• Journal of Periodontal and Implant Science
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• v.30 no.1
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• pp.91-104
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• 2000

Many synthetic bone materials have been studied for their potential of regenerative effects in periodontal tissue. Safflower seeds have been traditionally used as a drug for the treatment of fracture and blood stasis in oriental medicines. The purpose of this study was to assess and compare the osseous responses in rat calvarial defects between bone substitutes such as calcium carbonate and bovine-derived hydroxyapatite and feeding of safflower seeds. The calvarial defects were made with 8 mm trephine bur in 24 Sprague-Dawley rats. Two graft materials were implanted in each experimental groups, whereas the control and safflower seed feeding groups were sutured without any other treatment. And then the rats of safflower seed feeding group were supplied with 3 g/day of safflower seeds. Each group was sacrificed at 4 weeks and 8 weeks. To study a histopathology related to bone healing and regeneration, Goldner's Masson Trichrome stain was done at each weeks. The tissue response was evaluated under light microscope. There were more osteoblastic activity, new bone formation, dense bony connective tissues in bovine-derived hydroxyapatite group compared to other groups at 8 weeks. The osseous defect area of safflower seed feeding group was filled with prominent fibrous tissues, where less inflammatory infiltration and new capillary proliferation. In the early phase of bone healing, safflower seed feeding reduces the inflammatory response and promotes the proliferation of connective tissue. These results suggest that natural bovine-derived HA and safflower seed feeding could enhance the regenerative potential in periodontal defects.

• Environmental Mutagens and Carcinogens
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• v.24 no.1
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• pp.33-39
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• 2004

To validate and to estimate the chemical hazard playa very important role to environment and human health. The detection of many synthetic chemicals including agrochemicals that may pose a genetic hazard in our environment is of great concern at present. Since these substances are not limited to the original products, and enter the environment, they have become widespread environmental pollutants, thus leading to a variety of chemicals that possibly threaten the public health. Pyrazosulfuron-ethyl [Ethyl-5-(4,6-dimethoxypyrimidin-2-ylcarbamoylsulfamoyl)-1-methylpyrazole-4-carboxylate, $C_{14}H_{18}N{6}O_{7}S,$ M.W. =414.39, CAS No. 93697-74-6], is one of well known rice herbicide belong in the sulfonyl urea group. To clarify the genotoxicity of this agrochemical, Ames bacterial reversion assay, in vitro chromosomal aberration assay with Chinese hamster lung (CHL) fibroblast and bone marrow micronucleus assay in mice were subjected. In Ames assay, although pyrazosulfuron-ethyl revealed cytotoxic at 5,000-140 $\mug/plate$ in Salmonella typhimurium TA100, no dose-dependent mutagenic potential in 4.4～70 $\mug/plate$ of S. typhimurium TA 98, TA 100, TA1535 and TA 1537 both in the absence and presence of S-9 metabolic activation system was observed. Using CHL fibroblasts, the 50% cell growth inhibition concentration $(IC_{50})$ of pyrazosulfuron-ethyl was determined as 1,243 $\mug/mL,$ and no chromosomal aberration was observed both in the absence and presence of S-9 mixture in the concentration range of 311-1,243 $\mug/mL.$ And also, in vivo micronucleus assay using mouse bone marrow, pyrazosulfuron-ethyl revealed no remarkable induction of MNPCE (micronucleated polychromatic erythrocytes/1000 polychromatic erythrocytes) in the dose range of 625-2,500 mg/kg body weight when administered orally. Consequently, Ames bacterial gene mutation with Salmonella typhimurium, in vitro chromosome aberration with mammalian cells and in vivo bone marrow micronucleus assay revealed no clastogenic potential of pyrazosulfuron-ethyl in this study.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.35 no.1
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• pp.18-24
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• 2013

Purpose: This study is performed to determine the effects of titanium cap with various sizes of pores on bone formation during guided bone regeneration (GBR). Methods: Calvaria from 10 adult male rabbits were chosen as the recipient sites. A trephine bur with a diameter of 10 mm was used to form one round groove on each side of sagittal suture of the cranium, and a round bur with a diameter of 1.5 mm was used to form 6 small holes on the inner circles of round grooves to induce bleeding. In the control group, bone graft was not conducted, and closed titanium cap was fixed in the round groove. Bone graft was not performed in groups 1 and 2, but fixed on titanium caps with 0.2 mm, and 0.5 mm sized pores, respectively. For groups 3, 4, and 5, a synthetic bone graft material (${\beta}$-tricalcium phosphate, Cerasorb$^{(R)}$, Germany) was transplanted, and titanium caps without pore, with 0.2 mm and 0.5 mm sized pore were fixed, respectively. The animals were sacrificed 4 weeks after, and clinical, radiographical, and histomorphometrical evaluation of bone regeneration was performed. Results: In all groups, there were no clinical signs of infection, inflammation or wound dehiscence. Radiographic evaluation revealed well-defined semi-circular radiopacity inside the titanium cap of groups 3, 4, and 5. Histologically, the inner surface of the hemisphere was evenly lined with newly formed bone tissue, as well as grafted bone material in the group 3. In groups 4 and 5, the insertion of connective tissue was observed along the inner surface. However, the overall surface area between the grafts with different holes yielded no statistical significance in the histomorphometrical evaluation. Conclusion: Although the total area of newly formed bone showed no significant difference, excellent bone formation tendency was observed histologically when closed caps were used with bone graft was accompanied.