• 대한소아치과학회지
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• 제28권2호
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• pp.238-246
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• 2001

본 실험은 탁월한 골유도능으로 관심의 대상이 되고 있는 BMP의 세포내 신호 전달자로 알려진 Smad 1과 Smad 5가 조골세포 초기 분화표지인자인 ALP 유전자의 발현에 미치는 영향 및 그 조절기전을 알아보고자 하였다. BMP 처리 없이도 Smad에 의해 ALP가 발현되는가를 알아보기 위해 Smad 1과 Smad 5가 각각 stably transfection된 C2C12 세포를 3일간 배양후 histochemical assay를 하였고, Smad 1과 Smad 5의 expression vector와 ALP promoter vector를 transient co-transfection한 후 ALP promoter activity를 측정하였다. Smad에 의한 BMP의 효과를 알아보기 위해서 100ng/ml의 BMP-2를 처리한 군과 처리하지 않은 군으로 나누어 세포를 배양한후 ALP 유전자의 발현을 northern blot analysis로 확인 하였다. Smad가 ALP 유전자의 발현을 직접적으로 조절하는가를 알아보기 위해서는 단백질 합성억제제인 cycloheximide를 전처리하여 ALP 유전자의 발현을 northern blot analysis하였다. 이상의 실험결과 다음과 같은 결론을 얻었다. $\cdot$ Smad 1과 Smad 5가 과발현된 세포에서는 BMP 처리없이도 ALP가 발현된다. $\cdot$ Smad 1과 Smad 5가 과발현된 세포에서 BMP 처리후 ALP 발현 증가율이 대조군 보다 현저히 높게 나타나 Smad가 BMP 효과를 증가시킨다는 것을 알 수 있다. $\cdot$ Smad는 새로운 단백질의 합성을 통해 ALP 유전자를 발현시킨다.

• 대한치과보철학회지
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• 제51권2호
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• pp.82-89
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• 2013

연구 목적: 본 연구는 양극산화 임플란트에 골형성단백질과 혈관내피세포성장인자를 코팅한 임플란트를 토끼의 경골에 식립하여 골과 임플란트 계면의 골유착 향상의 가능성을 평가하고자 시행하였다. 연구 재료 및 방법: 6마리의 토끼 양측 경골에 코팅을 하지 않은 양극 산화 임플란트(대조군)와 골형성단백질과 혈관내피세포성장인자를 코팅한 임플란트(실험군)을 각각 한쪽에 2개씩 식립하였다, 토끼는 2주, 8주에 각각 3마리씩 희생하였고 전체 식립된 임플란트는 24개이었다. 각 시기별, 그룹별 임플란트 수는 각각 6개씩이었다. 임플란트안정지수(resonance frequency analysis (RFA)), 회전 제거력(Removable torque measurement (RTQ))을 희생 시기에 측정하였다. 독립표본 t-test (SPSS Ver. 15.0, Chicago, USA)을 이용하여 2주, 8주에서 대조군과 실험군의 차이를 비교 분석하고, 유의수준95%에서 통계적으로 검정하였다. 결과:대조군과 실험군 모두 8주에서 우수한 골유착을 보였다. 특히 실험군에서 8주에 ISQ, RTQ 값 모두 대조군과 비교하여 우수한 값을 나타내었다 (P<.05). 하지만 2주에서는 두군 사이에 통계적 유의성을 보이지 않았다(P>.05). 결론: 골형성 단백질과 혈관내피세포성장인자를 임플란트 표면에 코팅하여 식립하는 것은 치유의 후반기에 골유착을 증대하는 것으로 사료된다.

• 한방재활의학과학회지
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• 제28권1호
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• pp.1-17
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• 2018

Objectives The purpose of this study was to evaluate the effect of Jeopgolsan (JGS) extract on anti-oxidant, anti-inflammatory activities in RAW 264.7 cells and on factors related with fracture healing in skull fractured rat. Methods Experimental animals were divided into four groups: normal group without any treatment (Normal), contral group were treated orally with distilled water (Control), Experimental group were treated orally with JGS at a concentration of 200 mg/kg/day (JGS 200) and Experimental group were treated orally with JGS at a concentration of 200 mg/kg/day (JGS 400). Rats in each group except the normal group were induced fractures in the skull. The 1,1-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical scavenging activity were measured to evaluate antioxidant activity. The production of nitric oxide (NO), $interleukin-1{\beta}$ ($IL-1{\beta}$), interleukin-6 (IL-6) and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) in the RAW 264.7 cells were measured to evaluate anti-inflammatory activity. The production of osteocalcin calcitonin, carboxy-terminal telepeptides of type II collagen (CTX II), transforming growth $factor-{\beta}$ ($TGF-{\beta}$), bone morphogenetic protein-2 (BMP-2), Insulin and alkaline phosphatase (ALP) in serum of rats were measured to evaluate the effects of fracture healing at 0, 2, 4, and 6th week. X-rays were taken every 3 week from 0 to 6th week to evaluate fracture healing effect. Results 1. No cytotoxicity was observed. 2. DPPH and ABTS radical scavenging activity were increased in a concentration dependent manner, indicating anti-oxidant effect. 3. NO, $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ were not significantly changed, indicating no anti-inflammatory effect. 4. Osteocalcin, Calcitonin, $TGF-{\beta}$ and ALP were significantly increased in the experimental groups. 5. CTX II, insulin were significantly decreased in the expermental groups. 6. Radiologic examination showed that union of fracture was promoted. Conclusions From above results, JGS showed significant results in factors related with fracture healing and radiologic examination. Threfore, JGS is expected to be effective in the treatment of fracture.

• Animal Bioscience
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• 제34권4호
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• pp.546-557
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• 2021

Objective: If fertilization does not occur within a specific period, the quality of unfertilized oocytes in the oviduct (in vivo aging) or in culture (in vitro aging) will deteriorate over time. Icariin (ICA), found in all species of Epimedium herbs, has strong antioxidant activity, and is thought to exert anti-aging effects in vitro. We asked whether ICA protects oocytes against age-related changes in vitro. Methods: We analyzed the reactive oxygen species (ROS) levels and expression of antioxidant, maternal, and estrogen receptor genes, and along with spindle morphology, and the developmental competence and quality of embryos in the presence and absence of ICA. Results: Treatment with 5 μM ICA (ICA-5) led to a significant reduction in ROS activity, but increased mRNA expression of glutathione and antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, peroxiredoxin 5, and nuclear factor erythroid 2-like 2), during aging in vitro. In addition, ICA-5 prevented defects in spindle formation and chromosomal alignment, and increased mRNA expression of cytoplasmic maturation factor genes (bone morphogenetic protein 15, cyclin B1, MOS proto-oncogene, serine/threonine kinase, and growth differentiation factor-9). It also prevented apoptosis, increased mRNA expression of antiapoptotic genes (BCL2-like 1 and baculoviral IAP repeat-containing 5), and reduced mRNA expression of pro-apoptotic genes (BCL2 antagonist/killer 1 and activation of caspase-3). Although the maturation and cleavage rates were similar in all groups, the total cell number per blastocyst and the percentage of apoptotic cells at the blastocyst stage were higher and lower, respectively, in the control and ICA-5 groups than in the aging group. Conclusion: ICA protects oocytes against damage during aging in vitro; therefore, it can be used to improve assisted reproductive technologies.

• BMB Reports
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• 제45권9호
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• pp.509-514
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• 2012

We have found that the previously uncharacterized bone morphogenetic protein-9 (BMP9) is one of the most osteogenic factors. However, it is unclear if BMP9 cross-talks with $TGF{\beta}1$ during osteogenic differentiation. Using the recombinant BMP9 adenovirus, we find that low concentration of rh$TGF{\beta}1$ synergistically induces alkaline phosphatase activity in BMP9-transduced C3H10T1/2 cells and produces more pronounced matrix mineralization. However, higher concentrations of $TGF{\beta}1$ inhibit BMP9-induced osteogenic activity. Real-time PCR and Western blotting indicate that BMP9 in combination with low dose of $TGF{\beta}1$ potentiates the expression of later osteogenic markers osteopontin, osteocalcin and collagen type 1 (COL1a2), while higher concentrations of $TGF{\beta}1$ decrease the expression of osteopontin and osteocalcin but not COL1a2. Cell cycle analysis reveals that $TGF{\beta}1$ inhibits C3H10T1/2 proliferation in BMP9-induced osteogenesis and restricts the cells in $G_0/G_1$ phase. Our findings strongly suggest that $TGF{\beta}1$ may exert a biphasic effect on BMP9-induced osteogenic differentiation of mesenchymal stem cells.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권6호
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• pp.460-465
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• 2010

Introduction: A cleft palate is a common birth defect in humans with an incidence of 1/500 to 1/1,000 births. It appears to be caused by multiple genetic and environmental factors during palatogenesis. Many molecules are involved in palate formation but the biological mechanisms underlying the normal palate formation and cleft palate are unclear. Accumulating evidence suggests that transforming growth factor $\beta$/bone morphogenetic proteins (TGF-$\beta$/BMP) family members mediate the epithelial-mesenchymal interactions during palate formation. However, their roles in palatal morphogenesis are not completely understood. Materials and Methods: To understand the roles of TGF-$\beta$/BMP signaling in vivo during palatogenesis, mice with a palatal mesenchyme- specific deletion of Smad4, a key intracellular mediator of TGF-$\beta$/BMP signaling, were generated and analyzed using the Osr2Ires-Cre mice. Results: The mutant mice were alive at the time of birth with open eyelids and complete cleft palate but died within 24 hours after birth. In skeletal preparation, the horizontal processes of the palatine bones in mutants were not formed and resulted in a complete cleft palate. At E13.5, the palatal shelves of the mutants were growing as normally as those of theirwild type littermates. However, the palatal shelves of the mutants were not elevated at E14.5 in contrast to the elevated palatal shelves of the wild type mice. At E15.5, the palatal shelves of the mutants were elevated over the tongue but did not come in contact with each other, resulting in a cleft palate. Conclusion: These results suggest that mesenchymal Smad4 mediated signaling is essential for the growth of palatal processes and suggests that TGF-$\beta$/BMP family members are essential regulators during palate development.

• Reproductive and Developmental Biology
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• 제35권1호
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• pp.9-15
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• 2011

The functional cardiovascular system is comprised of distinct mesoderm-derived lineages including endothelial cells, vascular smooth muscle cells and other mesenchymal cells. Recent studies in the human embryonic stem cell differentiation model have provided evidence indicating that these cell lineages are developed from the common progenitors such as hemangioblasts and cardiovascular progenitor cells. Also, the studies have suggested that these progenitors have a common primordial progenitor, which expresses KDR (human Flk-1, also known as VEGFR2, CD309). We demonstrate here that sustained activation of BMP4 (bone morphogenetic protein 4) in hESC line, CHA15 hESC results in $KDR^+$ mesoderm specific differentiation. To determine whether the $KDR^+$ population derived from hESCs enhances potential to differentiate along multipotential mesodermal lineages than undifferentiated hESCs, we analyzed the development of the mesodermal cell types in human embryonic stem cell differentiation cultures. In embryoid body (EB) differentiation culture conditions, we identified an increased expression of $KDR^+$ population from BMP4-stimulated hESC-derived EBs. After induction with additional growth factors, the $KDR^+$ population sorted from hESCs-derived EBs displays mesenchymal, endothelial and vascular smooth muscle potential in matrix-coated monolayer culture systems. The populations plated in monolayer cultures expressed increased levels of related markers and exhibit a stable/homologous phenotype in culture terms. In conclusion, we demonstrate that the $KDR^+$ population is stably isolated from CHA15 hESC-derived EBs using BMP4 and growth factors, and sorted $KDR^+$ population can be utilized to generate multipotential mesodermal progenitors in vitro, which can be further differentiated into cardiovascular specific cells.

• Journal of Chest Surgery
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• 제36권2호
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• pp.86-90
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• 2003

폐 형성에 활성적인 또는 억제적인 기능을 갖고 있다고 알려져 있는 FGF-7 성장인자, BMP수용체 II, 그리고 TGF-$\beta$ 수용체 II 유전자의 비정상 발현이 폐기포 생성에 관여하는지를 각각의 단일성 클론 항체를 사용하여 수술로 절제된 자연기흉 환자의 폐기포 조직들을 면역조직염색 방법으로 염색하여 관찰하였다. 대상 및 방법: 재발성 또는 지속성 기흉으로 흉강경 또는 개흉술로 폐기포 절제술을 실시한 환자들을 대상으로 하였다. 총 31명의 환자로 15세에서 39세까지 연령분포를 보였으며 남자 30명, 여자 1명이었다. 폐기포 절제는 비디오흉강경이나 소절개개흠술을 통하여 폐기포벽에 손상을 가하지 않게 주의하면서 비디오흉강경용 스태플러(Endo GIA stapler)를 이용하여 절제하였으며 가능한 원형을 유지하여 신선한 상태로 포르마린에 고정하여 면역조직화학적 연구를 위한 표본을 만들었다. 폐기포 조직 슬라이드를 단일클론성 항 TGF-$\beta$ 수용체 II, BMP수용체 II 그리고 FGF-7인자 항체를 이용하여 면역조직학적 염색방법으로 관찰하였다. 결과: 전체 환자 31명중 TGF-$\beta$ 수용체 II항체에 양성 반응을 나타낸 환자수는 모두 24명이었다. 이들 중에는 18명이 강한 양성 반응을 보였고, 6명이 약한 양성 반응을 보였다 면역조직화학적 염색 결과를 고배율 현미경으로 살펴보면, TGF-$\beta$ 수용체 II의 염색이 기흉과 정상 폐조직 경계 부위에서 측히 강하게 염색됨이 관찰되었다. 이에 반하여, BMP수용체 II 그리고 FGF-7인자의 항체를 이용한 면역조직학적 염색 결과는 모든 환자의 조직들에서 음성으로 관찰되었다. 결론: 폐 조직이 형성될 때, 억제유전자의 역할을 담당하고 있다고 알려진 TGF-$\beta$ 수용체 II의 발현이 증가되면서 폐기포가 생성될 수 있다는 가능성을 제시하였다. 이번 결론은 면역조직학적 염색 실험 결과만으로 밝혀진 사실임으로 좀 더 체계적인 분자생물학적 인 연구가 요구된다.