• Journal of Periodontal and Implant Science
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• 제39권2호
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• pp.119-128
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• 2009

Purpose: The aim of this study was to evaluate the stemness of cells from canine dental tissues and bone marrow. Methods: Canine periodontal ligament stem cells (PDLSC), alveolar bone stem cells (ABSC) and bone marrow stem cells(BMSC) were isolated and cultured. Cell differentiations (osteogenic, adipogenic and chondrogenic) and surface antigens (CD146, STRO-1, CD44, CD90, CD45, CD34) were evaluated in vitro. The cells were transplanted into the subcutaneous space of nude mice to assess capacity for ectopic bone formation at 8 weeks after implantation. Results: PDLSC, ABSC and BMSC differentiated into osteoblasts, adipocytes and chondrocytes under defined condition. The cells expressed the mesenchymal stem cell markers differently. When transplanted into athymic nude mice, these three kinds of cells with hydroxyapatite /${\beta}$- tricalcium phosphate (HA/TCP) carrier showed ectopic bone formation. Conclusions: This study demonstrated that canine dental stem cells have stemness like bone marrow stem cells. Transplantation of these cells might be used as a therapeutic approach for dental stem cell-mediated periodontal tissue regeneration.

• International Journal of Oral Biology
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• 제31권2호
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• pp.53-65
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• 2006

Calcium concentration in the bone resorption lacunae is high and is in the mM concentration range. Both osteoblast and osteoclast have calcium sensing receptor in the cell surface, suggesting the regulatory role of high extracellular calcium in bone metabolism. In vitro, high extracellular calcium stimulated osteoclastogenesis in coculture of mouse osteoblasts and bone marrow cells. Therefore we examined the genes that were commonly regulated by both high extracellular calcium and $1,25(OH)_2vitaminD_3(VD3)$ by using mouse oligo 11 K gene chip. In the presence of 10 mM $[Ca^{2+}]e$ or 10 nM VD3, mouse calvarial osteoblasts and bone marrow cells were co-cultured for 4 days when tartrate resistant acid phosphatase-positive multinucleated cells start to appear. Of 11,000 genes examined, the genes commonly regulated both by high extracellular calcium and by VD3 were as follows; 1) the expression of genes which were osteoclast differentiation markers or were associated with osteoclastogenesis were up-regulated both by high extracellular calcium and by VD3; trap, mmp9, car2, ctsk, ckb, atp6b2, tm7sf4, rab7, 2) several chemokine and chemokine receptor genes such as sdf1, scya2, scyb5, scya6, scya8, scya9, and ccr1 were up-regulated both by high extracellular calcium and by VD3, 3) the genes such as mmp1b, mmp3 and c3 which possibly stimulate bone resorption by osteoclast, were commonly up-regulated, 4) the gene such as c1q and msr2 which were related with macrophage function, were commonly down-regulated, 5) the genes which possibly stimulate osteoblast differentiation and/or mineralization of extracellular matrix, were commonly down-regulated; slc8a1, admr, plod2, lox, fosb, 6) the genes which possibly suppress osteoblast differentiation and/or mineralization of extracellular matrix, were commonly up-regulated; s100a4, npr3, mme, 7) the genes such as calponin 1 and tgfbi which possibly suppress osteoblast differentiation and/or mineralization of extracellular matrix, were up-regulated by high extracellular calcium but were down-regulated by VD3. These results suggest that in coculture condition, both high extracellular calcium and VD3 commonly induce osteoclastogenesis but suppress osteoblast differentiation/mineralization by regulating the expression of related genes.

• International Journal of Industrial Entomology and Biomaterials
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• 제32권2호
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• pp.80-89
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• 2016

The effects of Cudrania tricuspidata (CT) extract on markers of osteoporosis were examined in ovariectomized rats. We classified 26 rats into five groups and provided a pellet chow diet and tap water throughout the 27-wk experimental period. During the last 15 wk, we added oral injections to each group as follows: sham-operated (SHAM, n=4) and ovariectomized-control (OVX, n=5) with distilled water, alendronate with 10 mg/kg/d of alendronate sodium (ALEN, n=5), CT (CT100, n=6) with 100 mg/kg/d of CT, and CT (CT300, n=6) with 300 mg/kg/d of CT. After the experimental period, blood, urine, and micro-CT images were assessed. The CT100 and OVX groups did not show any significant differences in urinary n-terminal telopeptide (NTx) (p<0.05 ), but with increases in CT concentration, the NTx level was slightly reduced. Serum osteocalcin was significantly higher in the CT groups than in all other groups (p<0.05 ). Notably, the serum calcium levels of all groups were within the normal range, but urinary calcium levels in the CT groups were significantly lower than the OVX group (p<0.05 ). In addition, the CT groups exhibited higher trabecular BMD than the OVX groups while showing similar BMD to the ALEN group (p<0.05 ). The Tb.Th of the ALEN group was lower than all other groups. Based on the overall analysis of results, CT prevented bone loss by inhibiting bone resorption and enhancing bone formation. Although alendronate showed a similar effect in preventing bone loss, it did so by solely inhibiting bone resorption, and its long-term use reportedly causes paradoxical effects such as hip fractures. Thus, for osteoporosis induced by ovariectomy, we conclude that CT extract is an effective natural treatment without severe side effects.

• Journal of Nutrition and Health
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• 제48권5호
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• pp.381-389
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• 2015

본 연구는 성장기 동물모델의 고과당 식이섭취가 골성장 (bone growth) 및 성숙 (bone maturation)에 미치는 영향을 연구하기 위해 수행되었다. 이를 평가하기 위해 생화학적 표지자 (biochemical markers), 경골의 구조학적 파라미터 (structural parameters) 및 골밀도 (BMD), 신장의 조직학적 변화를 측정하였고, 결과를 요약하면 다음과 같다. 1) 30% 고과당 섭취 (8주 동안) 및 30% 고과당 + 45 kcal% 고지방 섭취는 경골 내 해면골 (Tibia trabecular bone)의 부피비 (BV/TV), 해면골의 평균 개수 (Tb.N) 및 패턴요소 (Tb.Pf)를 향상시켰다. 2) 45 kcal% 고지방 섭취는 경골 내 피질골 (Tibia trabecular bone)의 부피를 향상시켰다. 3) 30% 고과당 섭취는 신장의 출혈 (interstitial bleeding), 공포화 변성 (vacuolation), 사구체 울혈 (glomerular congestion) 등의 병변을 유발하지 않았다. 4) 30% 고과당 섭취는 혈중 OC 농도를 낮추었고, 요 중 DPD농도에는 영향을 주지 않았다. 5) 8주 동안의 30% 고과당 섭취는 유의한 경향의 체중증가를 유도하였고, 45 kcal% 고지방 섭취는 유의적인 체중증가를 나타내었다. 본 연구 결과에 따르면 고과당의 섭취는 신기능의 손상 없이 성장기 해면골을 더욱 치밀하게 만드는 것으로 나타났다. 많은 선행연구들을 통하여 설탕이나 포도당의 과잉 섭취가 골 소실을 유발한다고 보고되어 왔으며, 본 연구를 포함한 이전 연구들을 종합해보면 고과당 섭취는 설탕이나 포도당 과잉 섭취가 유발하는 골 소실을 유발하지는 않는 것으로 보인다. 하지만 이러한 결과가 고과당 섭취가 골형성을 돕는다는 것을 의미하지는 않는다. 따라서 본 연구의 결과는 골의 형성 및 성숙에 중요한 시기인 청소년기에 안전한 과당섭취 가이드라인을 위한 초석을 다지는데 참고자료로서 사용될 수 있으며, 설탕이나 포도당 등을 과당으로 대체하여 섭취하는 것이 골 건강에 도움이 될 수 있을지는 향후 추가적인 연구가 필요할 것으로 보인다.

• 대한치과교정학회지
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• 제37권3호
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• pp.212-219
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• 2007

현재 교정용 미니 임플란트의 식립부위에 대한 연구는 주로 구치부 치근사이 공간에 집중되어 왔으며, 연조직에 관한 연구는 부족하다. 본 연구의 목적은 하악골에서 전치부와 구치부의 치근사이 피질골과 연조직 두께를 측정하여 해부학적인 지도를 만들고, 한국인의 평균 부착치은의 폭경을 제시하는데 있다. 연구를 위하여 자원한 남자 15명, 여자 15명 총 30명을 대상으로 하였으며, 평균연령은 27세 3개월이었다. 구강 내 인상을 채득한 후 석고모형을 만들어 0.5 mm두께의 평판으로 스텐트를 제작하였다. 스텐트의 중절치에서 제1대구치까지 치근사이 치은변연을 연결한 선에 방사선 불투과성의 표식을 부착하고 구강 내에 장착한 후 3차원 CT 영상을 얻고, 각 치근사이 치조정 하방에서 2, 4, 6, 8 mm부위의 피질골 두께를 측정하였다. 연조직 두께는 스텐트의 각 치근사이 치조정 하방에서 2, 4, 6, 8 mm부위에 구멍을 뚫고 구강 내에서 근관치료용 파일을 연조직에 통과시켜 측정하였다. 석고모형 상에서 중절치에서 제1대구치까지 각 치근중앙 부위에서 부착치은의 폭경을 측정하였다. 연구결과 하악골에서 전체대상자의 평균 피질골 두께는 $1.33{\pm}0.38mm$이었으며 전치부에서 구치부로 갈수록 두꺼워지고, 평균 연조직 두께는 $1.49{\pm}0.54mm$이었으며 치근단 쪽으로 내려가면서 두께가 변하지 않았다. 평균 전체 두께는 $2.82{\pm}0.70mm$이었으며 전치부에서 구치부로 갈수록 두꺼워졌다. 부착치은의 폭경은 전치부가 구치부보다 넓었다. 이상의 결과는 피질골과 연조직 두께, 부착치은의 폭경을 고려하여 미니 임플란트의 식립부위를 결정하는 것이 필요함을 시사하였다.

• 대한임상검사과학회지
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• 제48권4호
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• pp.371-377
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• 2016

탈회방법은 골수조직의 병리학적 진단을 위해서 항상 시행되는 과정이다. HCl 탈회용액과 같이 주로 사용하고 있는 산성용액은 탈회과정 동안에 조직내의 항원성에 손상을 입힌다. 특히, 골수조직 내의 RNA나 DNA에 심하게 손상을 준다. 따라서 조직의 항원성을 보존하기 위한 표준화된 탈회방법이 필요하다. 본 연구는 일반적으로 가장 많이 사용되는 HCl 기반의 상품화된 탈회용액과 직접 제조한 EDTA 탈회용액이 골수조직의 탈회과정에 어떤 영향을 미치는지 분석하였다. 환자로부터 채취된 73예의 골수생검조직을 HCl 탈회와 EDTA 탈회의 두 그룹으로 나누어 탈회과정을 진행하였다. 골수생검조직의 탈회과정 후 결과의 차이는 hematoxylin & eosin 염색과 reticulum 염색, Ki-67, CD20, CD138의 항체를 이용한 면역조직화학염색, DNA 추출 및 분석, in situ hybridization, IGH gene rearrangement 와 같은 분자병리검사를 시행하여 분석하였다. 일반적인 염색과 특수염색에서는 두 탈회용액간의 차이는 없었다. 또한 세포증식 표지자와 같은 세포막 혹은 세포질에서 발현되는 항체는 탈회용액간의 차이 없이 잘 염색되었다. 반면 HCl 탈회 용액에 처리한 후 핵 내 단백질인 Ki-67의 염색상은 현저히 불량한 것으로 관찰되었다. HCl 탈회용액과 비교하여 EDTA 탈회용액에서의 골수생검조직 내의 DNA와 RNA가 잘 보존되었음을 다양한 분자병리검사를 통해 확인할 수 있었다. 특히 HCl 탈회용액에 처리한 28예와 EDTA 탈회용액에 처리한 12예의 DNA의 순도와 농도을 비교한 결과 통계학적으로 유의한 수준으로 차이가 있음을 확인하였다. 이로써 EDTA 탈회용액이 조직 내의 항원성을 잘 유지시키며, 면역조직화학염색과 분자병리검사에 적합한 방법임을 확인 할 수 있었다.

• Journal of Periodontal and Implant Science
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• 제47권5호
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• pp.273-291
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• 2017

Purpose: Although static magnetic fields (SMFs) have been used in dental prostheses and osseointegrated implants, their biological effects on osteoblastic and cementoblastic differentiation in cells involved in periodontal regeneration remain unknown. This study was undertaken to investigate the effects of SMFs (15 mT) on the osteoblastic and cementoblastic differentiation of human osteoblasts, periodontal ligament cells (PDLCs), and cementoblasts, and to explore the possible mechanisms underlying these effects. Methods: Differentiation was evaluated by measuring alkaline phosphatase (ALP) activity, mineralized nodule formation based on Alizarin red staining, calcium content, and the expression of marker mRNAs assessed by reverse transcription polymerase chain reaction (RT-PCR). Signaling pathways were analyzed by western blotting and immunocytochemistry. Results: The activities of the early marker ALP and the late markers matrix mineralization and calcium content, as well as osteoblast- and cementoblast-specific gene expression in osteoblasts, PDLCs, and cementoblasts were enhanced. SMFs upregulated the expression of Wnt proteins, and increased the phosphorylation of glycogen synthase $kinase-3{\beta}$ ($GSK-3{\beta}$) and total ${\beta}-catenin$ protein expression. Furthermore, p38 and c-Jun N-terminal kinase (JNK) mitogen-activated protein kinase (MAPK), and nuclear $factor-{\kappa}B$ ($NF-{\kappa}B$) pathways were activated. Conclusions: SMF treatment enhanced osteoblastic and/or cementoblastic differentiation in osteoblasts, cementoblasts, and PDLCs. These findings provide a molecular basis for the beneficial osteogenic and/or cementogenic effect of SMFs, which could have potential in stimulating bone or cementum formation during bone regeneration and in patients with periodontal disease.

• Biomolecules & Therapeutics
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• 제18권4호
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• pp.386-395
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• 2010

Bone marrow-derived mesenchymal stem cells (BM-MSC) are a multipotent cell population that can differentiate into neuron-like cells. Previously it has been reported that murine BM-MSC can differentiate into neuron-like cells by co-treatment with a Rho-associated kinase (ROCK) inhibitor -Y27632 and $CoCl_2$. In this study, we compared several ROCK inhibitors for the ability to induce human BM-MSCs to differentiate into neuron-like cells in the presence of $CoCl_2$. Y27632 with high specificity for ROCK at 1-30 ${\mu}M$ was best at inducing neuronal differentiation of MSCs. Compared to HA1077 and H1152, which also effectively induced morphological change into neuron-like cells, Y27632 showed less toxicity even at 100 ${\mu}M$, and resulted in longer multiple branching processes at a wide range of concentrations at 6 h and 72 h post-induction. H89, however, which has less specificity by inhibition of protein kinase A, S6 kinase 1 and MSK1 with similar or greater potency, was less effective at inducing neuronal differentiation of MSCs. Simvastatin, which can inhibit Rho, Ras, and Rac by blocking the synthesis of isoprenoid intermediates, showed little activity for inducing morphological changes of MSCs into neuron-like cells. Accordingly, the expression patterns for neuronal cell markers,including ${\beta}$-tubulin III, neuron-specific enolase, neurofilament, and microtubule-associated protein, were consistent with the pattern of the morphological changes. The data suggest that the ROCK inhibitors with higher specificity are more effective at inducing neuronal differentiation of MSCs.

• Journal of Korean Neurosurgical Society
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• 제30권8호
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• pp.963-969
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• 2001

Objectives : Cord blood stem cells have been widely used as donor cells for bone marrow transplantation recently. These cells can give rise to a variety of hematopoietic lineages to repopulate the blood. Recent observations reveal that some bone marrow cells and bone marrow stromal cells(MSCs) can grow to become either neurons or glial cells. It is, however, unclear whether or not there exists stems cells which can differentiate into neurons in the blood during the early stages of postnatal life. Methods : Human cord blood stem cells were prepared from human placenta after full term delivery. To induce neuronal differentiation of stem cells, ${\beta}$-mercaptoethanol was treated. To confirm the neuro-glial characteristics of differentiated stem cells, immunocytochemical stain for NeuN, neurofilament, glial fibrillary acidic protein(GFAP), microtubule associated protein2(MAP2) was performed. RT-PCR was performed for detecting nestin mRNA and MAP2 mRNA. Results : We showed in this experiment that neuro-glial markers(NeuN, neurofilament, MAP2, GFAP) were expressed and axon-like cytoplasmic processes are elaborated in the cultured human cord blood stem cells prepared from new born placenta after full term delivery. Nestin mRNA was also detected in fresh cord blood monocytes. Conclusions : These results suggest that human cord blood derived stem cells may be potential sources of neurons in early postnatal life.

• Molecules and Cells
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• 제25권2호
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• pp.216-223
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• 2008

It has been reported that bone marrow (BM)-side population (SP) cells, with hematopoietic stem cell activity, can transdifferentiate into cardiomyocytes and contribute to myocardial repair. However, this has been questioned by recent studies showing that hematopoietic stem cells (HSCs) adopt a hematopoietic cell lineage in the ischemic myocardium. The present study was designed to investigate whether BM-SP cells can in fact transdifferentiate into functional cardiomyocytes. Phenotypically, BM-SP cells were $19.59%{\pm}9.00\;CD14^+$, $8.22%{\pm}2.72\;CD34^+$, $92.93%{\pm}2.68\;CD44^+$, $91.86%{\pm}4.07\;CD45^+$, $28.48%{\pm}2.24\;c-kit^+$, $71.09%{\pm}3.67\;Sca-1^+$. Expression of endothelial cell markers (CD31, Flk-1, Tie-2 and VEGF-A) was higher in BM-SP cells than whole BM cells. After five days of co-culture with neonatal cardiomyocytes, $7.2%{\pm}1.2$ of the BM-SP cells expressed sarcomeric ${\alpha}$-actinin as measured by flow cytometry. Moreover, BM-SP cells co-cultured on neonatal cardiomyocytes fixed to inhibit cell fusion also expressed sarcomeric ${\alpha}$-actinin. The co-cultured BM-SP cells showed neonatal cardiomyocyte-like action potentials of relatively long duration and shallow resting membrane potential. They also generated calcium transients with amplitude and duration similar to those of neonatal cardiomyocytes. These results show that BM-SP cells are capable of functional cardiomyogenic differentiation when co-cultured with neonatal cardiomyocytes.