• 디지털융복합연구
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• 제11권9호
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• pp.365-370
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• 2013

Image J 프로그램은 digital image 분석 또는 생명 공학적 분석에 사용되고 있다. 다양한 동물실험 중에서도 마우스 두개골 결손모델(Mouse calvarial defected model)은 조직공학자들에게 대표적으로 사용되고 있는 뼈의 결손모델이다. 이러한 마우스 두개골 결손 모델은 뼈의 재생 또는 새로운 지지체(scaffold)의 재생효과 실험에 사용되고 있다. 마우스 두개골에 지름 4mm의 원형 결손을 만들어 각종 실험재료를 처리하여 파괴된 마우스 두개골 뼈의 재생에 관련하여 측정하는 실험이 주를 이룬다. 실험 후에 마우스를 희생하여 대부분 헤마톡실린과 에오신 염색을 통하여 분석을 하게 된다. 다른 한편으로는 마이크로 시티(u-CT)를 사용하여 정량적인 분석을 한다. 하지만 이는 시간이 오래 걸리고 고가의 비용이 드는 단점이 있다. 이번 실험은 마우스 두개골 결손 모델 시에 프리쉐어 프로그램인 Image J를 통하여 간단하게 마우스 두개골 결손 뼈의 재생량을 2D X-ray 상으로 농도를 측정하여 정량적인 분석을 실행하였다. 결론적으로 Image J를 사용한 농도 분석법은 빠르고 정확하게 뼈의 재생량을 상대적으로 비교함으로써 신생 뼈의 관한 재생 실험자에게 도움을 줄 수 있을 것으로 사료된다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제39권3호
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• pp.120-126
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• 2013

Objectives: This study sought to elucidate the effect of autogenous tooth bone material by experimenting on minipig's maxillary sinus and performing histological and histomorphometric analyses. Materials and Methods: Five 18-24 month-old male minipigs were selected, and right maxillary sinuses were grafted with bone graft material made of their respective autogenous teeth extracted eight weeks earlier. The left sides were grafted with synthetic hydroxyapatite as control groups. All minipigs were sacrificed at 12 weeks after bone graft, which was known to be 1 sigma (${\sigma}$) period for pigs. Specimens were evaluated histologically under a light microscope after haematoxylin-eosin staining followed by semi-quantitative study via histomorphometric analysis. The ratio of new bone to total area was evaluated using digital software for calculation of area. Results: All specimens were available, except one on the right side (experimental group), which was missing during specimen preparation. This study demonstrated new bone at the periphery of the existing bone in both groups, showing evidence of bone remodeling, however, encroachment of new bone on the central part of the graft at the 1 ${\sigma}$ period was observed only in the autogenous tooth bone group (experimental group). Histomorphometric analysis showed more new bone formation in the experimental group compared to the control group. Although the difference was not statistically significant (P>0.05), the mean percentage area for new bone for the experimental and control groups were $57.19%{\pm}11.16%$ and $34.07%{\pm}13.09%$, respectively. Conclusion: The novel bone graft material using autogenous tooth is a good alternative to autogenous bone, comparable to autogenous bone, and outperforming synthetic hydroxyapatite bone graft materials in terms of bone regeneration capacity. Augmentation with autogenous tooth bone materials will reduce donor site morbidity without hampering the safety of the autogenous bone graft.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제38권
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• pp.27.1-27.9
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• 2016

Background: The objective of this study was to place bone graft materials in cranial defects in a rabbit model and compare their bone regenerating ability according to the size and density of demineralized dentin matrix (DDM). Methods: We selected nine healthy male rabbits that were raised under the same conditions and that weighed about 3 kg. Two circular defects 8 mm in diameter were created in each side of the cranium. The defects were grafted with DDM using four different particle sizes and densities: 0.1 mL of 0.25- to 1.0-mm particles (group 1); 0. 2 mL of 0.25- to 1.0-mm particles (group 2); 0.1 mL of 1.0- to 2.0-mm particles (group 3); and 0.2 mL of 1.0- to 2. 0-mm particles (group 4). After 2, 4, and 8 weeks, the rabbits were sacrificed, and bone samples were evaluated by means of histologic, histomorphometric, and quantitative RT-PCR analysis. Results: In group 1, osteoblast activity and bone formation were greater than in the other three groups on histological examination. In groups 2, 3, and 4, dense connective tissue was seen around original bone even after 8 weeks. Histomorphometric analysis of representative sections in group 1 showed a higher rate of new bone formation, but the difference from the other groups was not statistically significant. RT-PCR analysis indicated a correlation between bone formation and protein (osteonectin and osteopontin) expression. Conclusions: DDM with a space between particles of $200{\mu}m$ was effective in bone formation, suggesting that materials with a small particle size could reasonably be used for bone grafting.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권3호
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• pp.161-171
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• 2010

Introduction: This study was to assess the effectiveness of new bone formation and regeneration by using a rhBMP-2 and $\beta-TCP$ as a carrier in rabbits’mandible. Materials and Methods: The mandibles of 36 rabbits were exposed and cortical bone was penetrated for this study. The experimental subjects were divided into 3 groups each 12 rabbits ; control group, experimental group 1, and experimental group 2. Control group had the defect itself without any treatment, in the experimental group 1, $\beta-TCP$P only was grafted, and in the experimental group 2, rhBMP-2 soaked in $\beta-TCP$ was grafted. The rabbits were sacrificed after 1, 2, 3, 4, 6, and 8weeks, and new bone formation area was examined and measured for bone quantitative and qualitative analysis with light, fluorescent and polarized microscopy. Results: In the experimental group 1, new bone formation from the adjacent host bone was made by osteoconduction, and in the experimental group 2, direct new bone formation by osteoinduction of rhBMP-2 as well as new bone formation by osteoconduction of $\beta-TCP$ were observed. Conclusion: rhBMP-2 of experimental group 2 is very effective in the bone formation in early 2weeks and bone remodelling from 3weeks.

• Journal of Periodontal and Implant Science
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• 제33권3호
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• pp.359-372
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• 2003

Among objectives of periodontal therapy. the principal one is the morphological and functional reconstruction of lost periodontal supporting tissues. This includes de novo formation of connective tissue attachment and the regrowth of alveolar bone. The use of enamel matrix derivative(EMD) may be a suitable means of regeneration new periodontal attachment in the infrabony defects. Implant used to replace lost tooth but, implantitis occurred after installation. The purpose of this study was to investigate the effects of EMD on differentiation and growth of osteoblast in titanium disc. Twentyfive millimeter diameter and 1mm thick Ti disc which was coated 25, 50, 100, 200${\mu}g$/ml of EMD(Emdogain(R)) used as experimental group, 25, 50, 100, 200ng/d of rhBMP-2 as positive control group, and no coat as negative control group. A human osteosarcoma cell line Saos-2 was cultured in Ti disc and cell proliferation and Alkaline phosphatase (ALP) activity were measured at 1 and 6 days. PCR was performed at 2 and 8 hours. Semi-quantitative RT-PCR for mRNA expressions of various osteoblastic differentiation markers -type I collagen, ALP, osteopontin, and bone sialoprotein - were performed at appropriate concentrations based upon the results of MTT and ALP assay. Cultured cell-disc complexes were prepared for scanning electron microscopy (SEM) at 2 hour. Data were analyzed using Mann-Whitney and repeated- measures 1-way analysis of variance(SPSS software version 10,SPSS. Chicago. IL). After culture, there was more osteoblast in EMD100${\mu}g$/ml than in EMD50, 200${\mu}g$/ml on day 6. There was significant difference in experimental and positive control group compared control group, as times go by(1 and 6 days). Alkaline phosphatase activity was different significantly in EMD100, 200${\mu}g$/ml and BMP100, 200${\mu}g$/ml on day 6. The results of reverse transcriptase-polymerase chain reaction (RT-PCR) showed that expression of mRNA for ALPase, collagen type I, osteopontin. hone sialoprotein and BMP-2 was detected at 2 hour and 8 hour in EMI 200${\mu}g$/ml subgroup and BMP100ng/ml subgroup. The results of this study suggest that application of enamel matrix derivative on osteoblast attached to titanium surface facilitate the expression of bone specific protein and the differentiation and growth of osteoblast.