• International Journal of Oral Biology
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• 제33권1호
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• pp.33-43
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• 2008

In the process of bone remodeling, mineral phase of bone is dissolved by osteoclasts, resulting in elevation of calcium concentration in micro-environment. This study was performed to explore the effect of high extracellular calcium ($Ca{^{2+}}_e$) on mineralized nodule formation and on the expression of progressive ankylosis (Ank), plasma cell membrane glycoprotein-1 (PC-1) and osteopontin by primary cultured mouse calvarial cells. Osteoblastic differentiation and mineralized nodule formation was induced by culture of mouse calvarial cells in osteoblast differentiation medium containing ascorbic acid and ${\beta}$-glycerophosphate. Although Ank, PC-1 and osteopontin are well known inhibitors of mineralization, expression of these genes were induced at the later stage of osteoblast differentiation during when expression of osteocalcin, a late marker gene of osteoblast differentiation, was induced and mineralization was actively progressing. High $Ca{^{2+}}_e$(10 mM) treatment highly enhanced mRNA expression of Ank, PC-1 and osteopontin in the late stage of osteoblast differentiation but not in the early stage. Inhibition of p44/42 MAPK activation but not that of protein kinase C suppressed high $Ca{^{2+}}_{e^-}$induced expression of Ank, PC-1 and osteopontin. When high $Ca{^{2+}}_e$(5 mM or 10 mM) was present in culture medium during when mineral deposition was actively progressing, matrix calcifiation was significantly increased by high $Ca{^{2+}}_e$. This stimulatory effect was abolished by pyrophosphate (5 mM) or levamisole (0.1-0.5 mM), an alkaline phosphatase inhibitor. In addition, probenecid (2mM), an inhibitor of Ank, suppressed matrix calcification in both control and high $Ca{^{2+}}_{e^-}$treated group, suggesting the possible role of Ank in matrix calcification by osteoblasts. Taken together, these results showed that high $Ca{^{2+}}_e$ stimulates expression of Ank, PC-1 and osteopontin as well as matrix calcification in late differentiation stage of osteoblasts and that p44/42 MAPK activation is involved in high $Ca{^{2+}}_{e^-}$induced expression of Ank, PC-1 and osteopontin.

• Imaging Science in Dentistry
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• 제38권3호
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• pp.125-132
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• 2008

Purpose : To investigate the effects of irradiation on transforming growth factor ${\beta}_1$ (TGF-${\beta}_1$) mRNA expression and calcific nodule formation in MC3T3-E1 osteoblastic cell line. Materials and Methods : Cells were cultured in alpha-minimum essential medium ($\alpha$-MEM) supplemented with 10% fetal bovine serum and antibiotics. When the cells reached the level of 70-80% confluence, culture media were changed with $\alpha$-MEM supplemented with 10% FBS, 5 mM $\beta$-glycerol phosphate, and $50\;{\mu}g/mL$ ascorbic acid. Thereafter the cells were irradiated with a single dose of 2, 4, 6, 8 Gy at a dose rate of 1.5 Gy/min. The expression pattern of TGF-${\beta}_1$ mRNA, calcium content and calcific nodule formation were examined on day 3, 7, 14, 21, 28, respectively, after the irradiation. Results : The amount of TGF-${\beta}_1$ mRNA expression decreased significantly on day 7 after irradiation of 4, 6, 8 Gy. It also decreased on day 14 after irradiation of 6, 8 Gy. and decreased on day 21 after irradiation of 8 Gy. The amount of calcium deposition decreased significantly on day 7 after irradiation of 4, 8 Gy (P < 0.01) and showed a decreased tendency on day 14, 21 after irradiation of 4, 6, 8 Gy. The number of calcific nodules was decreased on day 7 after irradiation of 4, 8 Gy. Conclusion: Irradiation with a single dose of 4, 6, 8 Gy influences negatively the bone formation at the molecular level by affecting the TGF-${\beta}_1$ mRNA expression that was associated with proliferation and the production of extracellular matrix in MC3T3-E1 osteoblastic cell line.

• Journal of Periodontal and Implant Science
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• 제39권sup2호
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• pp.279-286
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• 2009

Purpose: Gene therapy (ex vivo) has recently been used as a means of delivering bone morphogenetic proteins (BMPs) to sites of tissue regeneration. In the present study, we investigated the effect of co-transduction of adenoviruses expressing BMP-2 and BMP-7 on osteogenesisof C2C12 cells in vitro. Methods: A replication-defective human adenovirus 5 (Ad5) containing a cDNA for BMPs in the E1 region of the virus (Ad5BMP-2 and Ad5BMP-7) was constructed by in vivo homologous recombination. Functional activity of Ad5BMP-2 and Ad5BMP-7 were evaluated in mouse stromal cells (W20-17cells). C2C12 cells are transduced with various MOI (multiplicity of infection) of Ad5BMP-2 and Ad5BMP-7 to assess most effective and stable titer. Based on this result, C2C12 cells were transduced with Ad5BMP-2 and Ad5BMP-7 alone or by combination. BMPs expression, alkaline phosphatase (ALPase) activity, cell proliferation, and mineralization were assessed. Results: Ad5BMP-2 and Ad5BMP-7 are successfully transduced to W20-17 cells, and secreted BMPs stimulated cell differentiation. Also, C2C12 cells transduced with Ad5BMPs showed expression of BMPs and increased ALPaseactivity. In all groups, cell proliferation was observed over times. At 7days, cells co-transduced with Ad5BMP-2 and Ad5BMP-7 showed lower proliferation than the others. C2C12 cells co-transduced with Ad5BMP-2 and Ad5BMP-7 had greater ALPaseactivity than that would be predicted if effect of individual Ad5BMPs were additive. Little mineralized nodule formation was detected in cells transduced with individual Ad5BMPs. In contrast, Ad5BMP-2 and Ad5BMP-7 combination stimulated mineralization after culturing for 10 days in mineralizing medium. Conclusions: Present study demonstrated that adenoviruses expressing BMPs gene successfully produced BMPs protein and these BMPs stimulated cells to be differentiated into osteoblastic cells. In addition, the osteogenic activity of Ad5BMPs can be synergistically increased by co-transduction of cells with Ad5BMP-2 and Ad5BMP-7.

• 생명과학회지
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• 제16권6호
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• pp.925-931
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• 2006

골조직은 골아세포, 파골세포, 골세포 등으로 구성되며, 골개조시 여러 인자가 세포증식, 분화, 활성화 및 골대사 조절에 관여한다. 이때 조골세포의 활성은 골형성에 중요하므로, 본 연구에서는 MC3T3-E1 조골세포주를 이용하여 해조류로 조제한 알칼리의 ENA-A 이온수가 조골세포의 증식과 분화활성에 미치는 영향을 조사하였다. 먼저 ENA-A 이온수가 조골세포의 성장에 미치는 영향을 MTT 검색법으로 조사 한 결과, ENA-A 이온수 2% 처리시 대조군과 비교하여 134% 증가하여 조골세포에 대해 높은 성장률을 보였다. 또한 ALP 효소 활성에 미치는 영향을 9일간 배양하여 그 변화를 측정하였다. 결과, 농도 1, 2, 4, 8% 처리시 112, 150, 188%까지 ALP 활성을 증가시켰다. ENA-A 이온수는 다시 ALP 효소 염색법과 Alizarin Red 염색으로 조골세포의 ALP활성유도, 분화와 석회화 형성능을 재확인하였으며 골기질 유전자의 발현의 변화도 확인하였다. 이에 본 연구결과를 통해 각종 무기질 성분을 함유한 ENA 이온수가 조골세포의 분화시에 hydroxyapatite 형성에 영향을 줌으로써 골다공증 예방에 효과가 있을것으로 생각하여 이를 보고하는 바이다.

• BMB Reports
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• 제47권8호
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• pp.463-468
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• 2014

Osteoblasts are specialized mesenchymal cells that are responsible for bone formation. In this study, we examine the role of GATA4 in osteoblast differentiation. GATA4 was abundantly expressed in preosteoblast cells and gradually down-regulated during osteoblast differentiation. Overexpression of GATA4 in osteoblastic cells inhibited alkaline phosphatase activity and nodule formation in osteogenic conditioned cell culture system. In addition, overexpression of GATA4 attenuated expression of osteogenic marker genes, including Runx2, alkaline phosphatase, bone sialoprotein, and osteocalcin, all of which are important for osteoblast differentiation and function. Overexpression of GATA4 attenuated Runx2 promoter activity, whereas silencing of GATA4 increased Runx2 induction. We found that GATA4 interacted with Dlx5 and subsequently decreased Dlx5 binding activity to Runx2 promoter region. Our data suggest that GATA4 acts as a negative regulator in osteoblast differentiation by downregulation of Runx2.

• 대한관절경학회지
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• 제2권1호
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• pp.72-76
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• 1998

Between March 1992 and December 1995 at St. Paul's, Holy Family, St. Vincent and Eui Jung Bu St. Mary Hospitals Catholic University, two hundred and eighty patients underwent arthroscopic anterior cruciate ligament(ACL) reconstruction using central one-third bone-patellar tendon-bone autograft. Nine of these patients had limitation of motion(LOM) defined as a knee flexion contracture greater than 10o or less than 125o of passive knee flexion. This study analyzes the causes of LOM after ACL reconstruction as well as the results after passive manipulation or arthroscopic adhesiolysis under anesthesia for LOM. The results are as follows: 1. Out of nine patients, initially three had isolated ACL injuries and six had combined injuries. Seven of nine cases were perfomed by ACL reconstruction within four weeks and two were performed after four weeks following injury. 2. Treatment for LOM after ACL reconstruction was done after 5.5 months on average. 3. Arthroscopic adhesiolysis was done in 5 cases. There were fibrous adhesions at suprapatellar pouch and femoral intercondylar notch in all cases, respectively, infrapatellar fat pad in 3 cases and medial para patellar gutter in 2 cases. Two patients had a fibrous nodule, "cyclops" lesion, which formed anterior to the ACL graft. 4. Range of motion and Lysholm knee score were much improved following passive manipulation or arthroscopic adhesiolysis under anesthesia for LOM.

• 치위생과학회지
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• 제15권4호
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• pp.488-494
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• 2015

치주인대세포의 효과적인 조절은 성공적인 치주 조직 재생에 중요한 역할을 한다. NFATc1의 활성화가 골모세포에서 분화를 자극하지만, 치주인대세포가 골모세포로 분화하는 과정에서 NFATc1의 역할은 아직 보고되지 않았다. 본 연구는 hPDLCs가 골모세포로 분화하는 동안 NFATc1의 mRNA의 발현과 단백질 발현이 유도됨을 처음으로 확인하였다. CsA에 의한 NFATc1의 억제는 세포증식을 감소시켰다. 게다가, CsA를 처리한 결과, 분화표지자, ALP activity 및 광화결정형성을 감소시켰다. 이러한 연구 결과는 NFATc1이 치주 재생을 위한 골모세포 분화에 중요한 조절자 역할을 할 수 있을 것으로 생각된다.

• Clinics in Shoulder and Elbow
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• 제15권2호
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• pp.138-142
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• 2012

색소 융모 결절성 활액막염(Pigmented villonodular synovitis)은 활액막, 건막 및 점액낭 등에 융모 증식과 결절을 형성하는 드문 양성 활액막 증식성 질환으로 대부분 성인의 슬관절과 고관절에 발생한다. 수부, 완관절, 견관절 및 주관절에는 드물게 발생하며, 특히 소아에서의 주관절 색소 융모 결절성 활액막염은 아주 드물다. 좌측 주관절 통증이 있는 8세 남아의 단순 주관절 방사선 검사상 좌측 주두와에 양성 골종양과 같은 병변이 관찰되었다. 수술시 좌측 주관절의 활액막 이상 증식 소견이 관찰되어 임상적으로 색소 융모 결절성 활액막염을 의심하였으며, 병변의 개방 소파술 및 좌측 주관절의 근치적 활액막 절제술을 시행하였다. 절제 조직의 조직학적 검사 결과 색소 융모 결절성 활액막염으로 확진 되었다. 술 후 좌측 주관절의 통증이 호전되었으며, 정상 운동범위를 회복하였다. 저자들은 소아의 주관절에 발생한 골종양을 닮은 색소 융모 결절성 활액막염 1예를 보고함과 동시에 문헌 고찰을 하였다.

• 대한골관절종양학회지
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• 제12권1호
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• pp.78-82
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• 2006

Kimura씨 병은 정확한 병인이 알려지지 않은 매우 드문 만성 염증성 질환으로 알려져 왔다. 주로 단독 또는 다발성으로 피하층에 결절 형태로 발생하며 대부분 양성인 경과를 취한다. 주로 경부에 주변 림파선 증식을 동반하여 발생한다. 조직학적으로 림프양 조직이 증식되어 있고, 호산구가 많은 염증 세포의 침윤과 모세 세정맥이 풍부한 병리 소견을 보인다. 치료방법으로는 스테로이드 치료, 방사선 치료 및 수술적 치료 등이 있다. 저자들은 상지에 발생한 2예의 Kimura씨 병을 경험하였기에 문헌 고찰과 함께 보고하는 바이다.

• 치과방사선
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• 제27권1호
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• pp.263-271
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• 1997

The authors diagnosed a 54-year-old male as central mucoepidennoid carcinoma after undergoing clinical, radiological and histopathological examinations. The characteristics were as followed : 1. Clinically, the patient complained of the painless unilateral swelling of the left mandibular molar region and had a pus discharge through the fistula. Painful nodule was palpated on the scalp of the left frontal area and it was regarded as a metastatic lesion. 2. Plain radiographs showed the ill-defined permeative radiolucent lesion. The osteolytic lesions were also detected in the cranial bone, number 9 and 11 ribs, scapula, and vertebral bodies. 3. The mandibular CT and PNS MRI showed the swelling of the left mandible and the enlargement of the several lymph nodes of 1.5 cm in size. 4. Histopathologically, many solid epidermoid type cells were mixed with mucus-secreting cells and they were arranged in duct-like structure. Most of them were epidermoid type, which indicates a high grade tumor. Mucins could be found in mucicamrine staining.