• Maxillofacial Plastic and Reconstructive Surgery
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• 제39권
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• pp.8.1-8.8
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• 2017

Background: For an effective bone graft for reconstruction of the maxillofacial region, an adequate vascular network will be required to supply blood, osteoprogenitor cells, and growth factors. We previously reported that the secretomes of bone marrow-derived mesenchymal stem cells (MSC-CM) contain numerous growth factors such as insulin-like growth factor (IGF)-1, transforming growth factor $(TGF)-{\beta}1$, and vascular endothelial growth factor (VEGF), which can affect the cellular characteristics and behavior of regenerating bone cells. We hypothesized that angiogenesis is an important step for bone regeneration, and VEGF is one of the crucial factors in MSC-CM that would enhance its osteogenic potential. In the present study, we focused on VEGF in MSC-CM and evaluated the angiogenic and osteogenic potentials of MSC-CM for bone regeneration. Methods: Cytokines in MSC-CM were measured by enzyme-linked immunosorbent assay (ELISA). Human umbilical vein endothelial cells (HUVECs) were cultured with MSC-CM or MSC-CM with anti-VEGF antibody (MSC-CM + anti-VEGF) for neutralization, and tube formation was evaluated. For the evaluation of bone and blood vessel formation with micro-computed tomography (micro-CT) and for the histological and immunohistochemical analyses, a rat calvarial bone defect model was used. Results: The concentrations of IGF-1, VEGF, and $TGF-{\beta}1$ in MSC-CM were $1515.6{\pm}211.8pg/mL$, $465.8{\pm}108.8pg/mL$, and $339.8{\pm}14.4pg/mL$, respectively. Tube formation of HUVECs, bone formation, and blood vessel formation were increased in the MSC-CM group but decreased in the MSC-CM + anti-VEGF group. Histological findings suggested that new bone formation in the entire defect was observed in the MSC-CM group although it was decreased in the MSC-CM + anti-VEGF group. Immunohistochemistry indicated that angiogenesis and migration of endogenous stem cells were much more abundant in the MSC-CM group than in the MSC-CM + anti-VEGF group. Conclusions: VEGF is considered a crucial factor in MSC-CM, and MSC-CM is proposed to be an adequate therapeutic agent for bone regeneration with angiogenesis.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권4호
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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.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제31권5호
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• pp.365-374
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• 2009

Purpose : This research evaluates the effect of the use of absorbable membrane barrier with deproteinized bovine bone (Bio-$Oss^{(R)}$, Switzerland) on bone healing in surgically created critical-sized defects in rat calvaria. Materials and Methods : Two standardized transosseous circular calvarial defects (5 mm in diameter) are made in each calvarium of 30 rats. These rats are divided into negative control group(n=15), positive control group(n=15) and two experimental groups(n=15). In the negative control group, defects are only filled with blood clots. In the positive control group, defects are filled with autogenous bone obtained from calvarium; in the experimental group 1, defects are filled with deproteinized bovine bone; and in the experimental group 2, defects are filled with deproteinized bovine bone with absorbable membrane. At the postoperative 1 week, 3 weeks. and 6 weeks, clinical. histologic and histomorphometric evaluations of the defects are performed. Results : 1. The grafted bone without membrane in the calvarial bone defect was scattered but, the grafted bone with membrane was stable. 2. $BioMesh^{(R)}$ membrane was absorbed beginning at 3 weeks, and was absorbed considerably at 6 weeks while maintaining the structural form of the membrane. 3. The use of membrane blocked soft tissue invasion. 4. In histomorphometric analysis. it showed the greatest amount of new bone formation in the positive control group. The amount of new bone formation was greater in the experimental group 2 than experimental group 1. At 6 weeks. the amount of new bone formation was greater in the positive control group than experimental group l(p<0.005). Conclusion : These results suggest that membrane increase the stability of grafted bone and protects from soft tissue invasion, and the use of the membrane may promote new bone formation in deproteinized bovine bone graft area.

• 대한치과보철학회지
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• 제48권1호
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• pp.16-27
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• 2010

연구 목적: 본 연구의 목적은 실험용 쥐의 초기 골 재생 과정에서 혈소판 농축 혈장 (platelet-rich plasma; PRP)이 난소호르몬 분비 유무에 따라 각각 어떤 효과를 나타내는지 확인 비교해 보는 것이다. 연구 재료 및 방법: 실험용 쥐 40마리 중 20마리에는 난소절제술 (ovariectomy; OVX)을 시행하여 골다공증을 유발시킨 상태에서 골 이식을 하였고, 나머지 20마리에는 난소절제술 없이 골 이식을 하였다. 또, 난소절제술을 시행한 쥐 중 10마리와 난소절제술을 시행하지 않은 쥐 중 10마리에는 골 이식 시 골전도성 합성골 이식재인 $MBCP^{TM}$ (Micro-& macro-porous biphasic calcium phosphate)에 혈소판 농축 혈장을 첨가하여 적용하고, 아래와 같은 실험군으로 구분하였다. A군; 10마리의 non-OVX 쥐/골 이식재 ($MBCP^{TM}$). B군; 10마리의 non-OVX 쥐/골 이식재($MBCP^{TM}$)+PRP. C군; 10마리의OVX 쥐/골 이식재($MBCP^{TM}$). D군; 10마리의OVX 쥐/골 이식재($MBCP^{TM}$)+PRP. 모든 실험동물의 두 개관 정중부에 직경 8 mm 원형의 임계 크기 결함을 한 개씩 인위적으로 형성한 후, 임계 크기 결함 내부에 골 이식재 및 혈소판 농축 혈장을 적용하여 골 이식술을 실시하였다. 골이식 시행 4주 후에 실험 동물을 희생시켜 표본을 제작한 후, 광학현미경상을 관찰하고 기존에 형성한 임계 크기 결함 내부에 새롭게 침착된 신생골의 면적을 측정하여 그 측정값을 통계 분석하였다. 결과: 신생골 면적 측정값을 각 군 간 비교하여 다음과 같은 결과를 얻었다. 1. 난소절제술이 시행되지 않은 정상 쥐에서는 혈소판 농축 혈장의 사용이 골 재생에 유의한 효과를 나타내지는 않았다 (p>.05). 2. 난소절제술이 시행된 골다공증 유발 쥐에서는 혈소판 농축 혈장의 사용이 골 재생에 유의한 효과를 나타내었다 (P<.05). 3. 혈소판 농축 혈장이 사용되지 않은 경우, 난소절제술이 시행된 골다공증 유발 쥐는 정상 쥐보다 골 재생 능력이 유의할 만큼 감소하였다 (P<.05). 4. 혈소판 농축 혈장이 사용된 경우, 난소절제술이 시행된 골다공증 유발 쥐라고 하더라도 정상 쥐보다 골 재생 능력이 유의할 만큼 감소하지는 않았다 (P>.05). 결론: 이상의 결과를 토대로, 골전도성 합성골 이식재와 함께 쓰인 혈소판 농축 혈장은 정상 쥐에서보다 난소절제술이 시행된 골다공증 유발 쥐에서 골 재생 및 치유에 더 큰 효과가 있음을 알 수 있었다.

• Journal of Periodontal and Implant Science
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• 제27권1호
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• pp.129-150
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• 1997

The purpose of this study was to evaluate the effects of tetracycline(TC}, flurbiprofen, and PDGF-BB loaded biodegradable membranes on the cell-attachment, the activity of loaded PDGF-BB, in vivo release kinetics, and guided bone regenerative potentials. To evaluate the cell attachment to membranes, the number of gingival fibroblasts attached to each membrane(10% TC, 10% flurbiprofen, $200ng/cm^2$ PDGF-BB loaded membranes, drug-unloaded membrane) was counted by coulter counter and the morphologic pattern of attached cells was examined under SEM. To determine whether the activity of loaded PDGF-BB is sustained, the cellular growth and survival rate of gingival fibroblasts was used for both standard PDGF-BB and loaded PDGF-BB. For evaluation of in vivo release kinetics, drug-loaded membranes were implanted on the dorsal skin of the rats. On 1, 3, 7, 10, 14, 21, and 28 days after implantation, the amount of remaining drugs were measured by HPLC assay for TC and flurbiprofen, and by ${\gamma}-scintillation$ counter for $PDGF-BB^{1125}$. For evaluation of guided regenerative potential, the amount of new bone in the calvarial defect(5mm in diameter) of the rat was measured by histomorphometry 1 and 2 weeks after implantation of membranes. The number of cells attached to the PDGF-BB loaded membrane was largest as compared with the other mernbranes.(p< 0.05) The activity of loaded PDGF-BB was not significantly different from the activity of standard PDGF-BB.(p<0.05) After initial burst release of drug during the first 24 hours, drugs were gradually released for 4 weeks. Especially the release rate of PDGF-BB was nearly constant during 4 weeks. PDGF-BB loaded membranes(200, $400ng/cm^2$) were effective in guided bone regeneration as compared with drug-unloaded membrane. These results implicate that drug-loaded biodegradable membranes might be a useful for guided bone regeneration.