• 생명과학회지
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• 제20권4호
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• pp.607-613
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• 2010

솔잎은 항산화력이 우수한 것으로 알려져 있으며, mouse calvaria 유래의 MC3T3-E1 osteoblastic cell은 골세포의 세포 활성과 관련된 연구에서 유용하게 이용되어왔다. 따라서 본 연구에서는 MC3T3-E1 세포를 이용하여 적송잎 용매별 추출물이 세포 증식에 미치는 영향과 ALP 활성 및 조골세포의 골 형성을 위한 필수 인자인 콜라겐 합성의 영향에 대해 검토하였다. 적송잎 추출물의 농도(1, 10 50, $100\;{\mu}g/ml$)에 따른 조골세포 성장에 미치는 영향을 MTT assay로 분석한 결과, proanthocyanidin의 경우 $50\;{\mu}g/ml$ 이상 첨가한 군에서 대조군보다 급격한 증식률을 나타내었다. 에탄올 추출물을 첨가하였을 때 유의적인 증식률이 나타나지 않은 반면, 적송잎 헥산 추출물 처리군에 있어서는 $10\;{\mu}g/ml$ 농도 이상에서 유의적으로 세포증식이 촉진되었다. 적송잎 추출물이 ALP 활성에 미치는 영향을 조사한 결과, 적송잎 추출물이 조골세포의 ALP의 활성을 증가시켜 조골세포의 분화에 영향을 줄 가능성이 제시 되었다. 또한 적송잎의 열수 및 에탄올 추출물보다 헥산 추출물에서 조골세포의 ALP 활성이 증가하여 적송잎의 ALP 활성에 영향을 주는 성분은 수용성 성분보다 지용성 성분인 것으로 추측되었다. 적송잎 추출물이 조골세포의 콜라겐 합성에 미치는 실험결과에서 헥산 추출물뿐만 아니라 열수 추출물에서도 높은 콜라겐 합성능력을 나타내었다. 따라서 단일성분에 의한 것보다 복합적 성분들의 상승작용에 의해 조골세포의 콜라겐 합성이 촉진된 것으로 추측할 수 있다. 적송잎 추출물이 조골세포의 증식, ALP 활성 및 콜라겐 합성을 촉진하여 골 생성에 영향을 줄 수 있는 것이 확인되었으며, 그에 대한 구체적인 기작 연구를 위하여 향후 분자생물학적인 차원에서의 in vitro 및 in vivo 실험을 통한 구체적인 연구가 수행되어야 할 것으로 사료된다.

• BMB Reports
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• 제42권7호
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• pp.427-432
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• 2009

Orthodontic tooth movement results from the combinational process of both bone resorption and formation in the compressive and tension sides, respectively. However, the genes responsible for new bone formation in tension sides have not been determined. In this study, we used DNA microarray and real-time RT-PCR to identify genes in human periodontal ligament (PDL) cells that undergo significant changes in expression in response to static tensional forces (2 or 12 hours). The genes found were alkaline phospatase (ALP), matrix metalloproteinases (MMPs), vascular endothelial growth factor (VEGF), and several collagen genes. Furthermore, an ELISA evaluating the expression of VEGF, type IV collagen and MMP-2 found levels significantly increased after 24 and 72 hours (P < 0.05). ALP activity was also increased after 24 hours (P < 0.05). Collectively, we found the genes up-regulated in our study by the static tensional force are related to osteogenic processes such as matrix synthesis and angiogenesis.

• Journal of Periodontal and Implant Science
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• 제35권4호
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• pp.1097-1108
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• 2005

The present study was to determine the influence of micro-macro biphasic calcium phosphate(MBCP) on proliferation and differentiation of human marrow-derived mesenchymal stem cells. Primary stem cells were cultured from bone marrow and 3-4 passaged cells were used. This study tested the proliferative effects by cell counting. Collagen sythensis, alkaline phosphatase activity, expression of osteocalcin and bone sialoprotein by Western blot analysis were evaluated. The cellular proliferation of ASC was not influenced by MBCP. Collagen synthesis of ASC cultured on MBCP significantly increased at 5th and 7th days(p<0.05). The ALP activity in ASC cultured on MBCP significantly increased at 5th and 7th days(p<0.05). The expression of OC and BSP incresaed in ASC cultured on MBCP. These results suggest that MBCP may stimulates the osteoblastic activity of ASC.

• BMB Reports
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• 제45권10호
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• pp.571-576
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• 2012

Radiotherapy is considered to cause detrimental effects on bone tissue eventually increasing bone loss and fracture risk. However, there is a great controversy on the real effects of irradiation itself on osteoblasts, and the mechanisms by which irradiation affects osteoblast differentiation and mineralization are not completely understood. We explored how X-ray radiation influences differentiation and bone-specific gene expression in mouse calvarial osteoblasts. Irradiation at 2 Gy not only increased differentiation and mineralization of the cells, but also upregulated the expression of alkaline phosphatase, type I collagen, osteopontin, and osteocalcin at early stages of differentiation. However, irradiation at higher doses (>2 Gy) did not stimulate osteoblast differentiation, rather it suppressed DNA synthesis by the cells without a toxic effect. Additional experiments suggested that transforming growth factor-beta 1 and runt-transcription factor 2 play important roles in irradiation- stimulated bone differentiation by acting as upstream regulators of bone-specific markers.

• 대한치과교정학회지
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• 제35권4호
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• pp.262-274
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• 2005

본 연구는 치주인대 세포에 지속적이고 점진적 인장력을 가하여 치아 이동 시 형성되는 인장부위의 기계적 자극에 대한 생화학적 전달과 치조골 흡수와 생성 조절 기전을 이해하고자 하였다 치주인대 세포가 배양된 유연한 성장 표면을 가진 배지에 지속적이고 점진적인 인장력을 가하고 골흡수 인자인 $PGE_2$와 골형성 인자인 ALP의 생성량을 1 3 5. 12시간 후에 측정하여 정량비교하였고 파골세포 분화기전을 조절하는 OPG RANKL의 인자들과 matrix metalloproteinase(MMP)-1, -8, -9, -13, tissue inhibitor of matrix metalloproteinase(TIMP)-1의 인자들을 역전사 중합효소 연쇄반응 검사하여 m-RNA 발현을 비교한 결과 치주인대 세포에 인장력을 가한 경우 대조 군보다 $PGE_2$의 농도가 적었고 (p<0.05) ALP의 농도 변화는 없었으며 OPG의 mRNA 발현이 증가하였으나, RANKL의 mRNA 발현은 감소하였다 그리고 TIMP-1과 MMP-1 -8 -9, -13의 mRNA 발현이 대조군과 차이가 없었다. 이상의 연구에서 사람의 치주인대 세포는 점진적이고 지속적인 인장력에 대한 반응으로 $PGE_2$의 생성과 RANKL의 mRNA 발현은 감소하고 OPG의 mRNA 발현은 증가하여 골흡수를 억제하는 효과를 보이는 것으로 나타났다.

• 대한한의학회지
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• 제27권1호
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• pp.220-228
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• 2006

Objectives : Interleukin-1 (IL-1)${\beta}$ in articular chondrocytes regulates differentiation, apoptosis, and inflammatory responses. It is still controversial, So, we investigated IL- $1{\beta}$ induces chondrocytes dedifferentiation and death. Also, we studied the role of the mitogen-activated protein kinase (MAPK) subtypes on IL-$1{\beta}$-induced dedifferentiation and apoptosis. Methods : To evaluation of dedifferentiation by chemokines of chondrocytes, we assessed such as proteoglycan, collagen, MMP-3 and MMP-13 by RT-PCR analysis. Also, to assess of apoptosis effect by chemokines, we measured annexin V/propidium iodode (PI) and sub G1 cells in chondrocytes by flowcytometric analysis Results : IL-$1{\beta}$ treatment did not affect activation of ERK-1/2, but stimulation of p38 kinase. Inhibition of phospho ERK-1/2 with PD98059 enhanced IL-1b-induced dedifferentiation, and apoptosis up to 13.5%, whereas inhibition of phospho p38 kinase with SB203580 inhibited dedifferentiation, and apoptosis. Conclusions : Our results indicate that SB203580, p38 kinase inhibitor, inhibits IL-$1{\beta}$-induced dedifferentiation, and apoptosis by the inhibition of type II collagen expression and proteoglycan synthesis of rabbit articular chondrocytes.

• Biomolecules & Therapeutics
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• 제20권3호
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• pp.299-305
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• 2012

Endochondral bone formation is the process by which mesenchymal cells condense to become chondrocytes, which ultimately form new bone. The process of chondrogenic differentiation and hypertrophy is critical for bone formation and as such is regulated by many factors. In this study, we aimed to indentify novel factors that regulate chondrogenesis. We investigated the possible role of isopsoralen in induction of chondrogenic differentiation in clonal mouse chondrogenic ATDC5 cells. Isopsoralen treatment stimulated the accumulation of cartilage nodules in a dose-dependent manner. Further, ATDC5 cells treated with isopsoralen were stained more intensely with Alcian blue than control cells, suggesting that isopsoralen increases the synthesis of matrix proteoglycans. Similarly, isopsoralen markedly induced the activation of alkaline phosphatase activity compared with control cells. Isopsoralen enhanced the expressions of chondrogenic marker genes such as collagen II, collagen X, OCN, Smad4 and Sox9 in a time-dependent manner. Furthermore, isopsoralen induced the activation of extracellular signal-regulated kinase (ERK) and p38 MAP kinase, but not that of c-jun N-terminal kinase (JNK). Isopsoralen significantly enhanced the protein expression of BMP-2 in a time-dependent manner. PD98059 and SB 203580, inhibitors of ERK and p38 MAPK, respectively, decreased the number of stained cells treated with isopsoralen. Taken together, these results suggest that isopsoralen mediates a chondromodulating effect by BMP-2 or MAPK signaling pathways, and is therefore a possible therapeutic agent for bone growth disorders.

• Preventive Nutrition and Food Science
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• 제1권1호
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• pp.80-86
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• 1996

Parathyroid hormone(PTH) is known to stimulate bone resorption and to inhibit bone collagen synthesis. In contrast, as the evidence of stimulation of bone formation by PTH has recently been observed, the study on the role of PTH involved in osteoporosis draws remarkable attention. This study has dealt with the role of alkaline phoshatase(AP), a marker enzyme for bone formation and osteoblast action, Animals(BALS/cmice) were divided into three dietary groups(high and medium Ca and Ca-free) and hormones including PTH, calcitonin(CT), cholecalciferol(citamin D) were i.p. injected. AP in the serum and liver was measured using Sigma 221 alkaline buffer solutions containing 9mM of p-nitrophenyl phoshate. Enzume was reacted at 37$^{\circ}C$ for 10 minutes and the reaction was stopped by 1.8ml of 0.1N NaOH and measured at 410nm. We found that serum and liver AP activity was increased by low dietary Ac. Compared to the control, and serum Ap activity was enhanced by PTH and vitamin D regardless of the dietary Ca. On the other hand, liver AP activity was inhibited by OTH and vitamin D at all levels of dietary Ca. CT inhibited the action of PTH and vitamin D in the serum. But, the inhibition of PTH and vitamin D action by CT was not observed in the liver, unlike in the case of serum.

• 대한치과교정학회지
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• 제20권3호
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• pp.499-517
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• 1990

Electrical stimulation among several factors that influence bone remodeling has been studied by many investigators with great enthusiasm in orthodontic field. The action mechanisms of Pulsed Electromagnetic Field (PEMF) are different from those of the conventional electrode application method in that PEMF induces endogenous current in the living tissues. PEMF is known to have the healing effect in nonunion of bone and osteoporosis. It is widely used in orthopaedic scopes and the possibility of using the method in clinical orthodontics Is also conceivable. But the exact mechanisms by which the PEMF exerts its effects are not clearly understood. Therefore, the author wanted to see the effect of PEMF on five groups of rat calvarial cells obtained by sequential enzyme digestion method, and observed the changes in enzyme activation, collagen synthesis and $^3H-thymidine$ incorporation. The results were as follows: 1. Under the effect of PEMF, there were no changes in the alkaline phosphatase activity in five groups of cell populations. 2. Both the PEMF group and the PTH with PEMF group shelved no changes in acid phosphatase activities and there were no differences between two experimental groups. 3. Under the effect of PEMF, there was significant increase of collagen synthesis in the group V cell population. 4. Under the effect of PEMF, there were significant increases of $^3H-thymidine$ incorporation in the group IV and V cell populations.

• 폴리머
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• 제37권6호
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• pp.669-676
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• 2013

탈미네랄화된 골분(demineralized bone particle, DBP)은 연골 형성의 유도인자로 사용되기 때문에 조직공학에서 널리 사용되는 생체재료이다. 본 연구에서는 in vivo 환경에서 디스크 재생 효과를 연구하기 위해 DBP를 poly(lactic-co-glycolic acid) (PLGA)에 첨가하여 다공성 지지체를 제조하였다. 디스크의 섬유륜 조직을 반으로 절개한 후, 수핵 조직을 제거하여 디스크 결손을 유발시켰다. 빈 공간에 PLGA, DBP/PLGA 지지체를 이식하여 in vivo 환경에서 조직공학적 디스크 재생을 관찰하였다. 1, 2 및 3개월 후 디스크를 적출하여 글리코스아미노글라이칸(glycosaminoglycan, sGAG) 및 콜라겐 합성 정도를 측정하였으며 조직학적 평가로 H&E, Safranin-O, Alcian blue 염색과 면역조직학적 평가로 제 I형 콜라겐, 제 II형 콜라겐 염색을 수행하였다. 그 결과 DBP/PLGA 지지체에서 sGAG 및 콜라겐 함량이 높은 것을 확인하였으며 추간판 디스크 재생의 가능성을 확인하였다.