• BMB Reports
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• 제49권10호
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• pp.548-553
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• 2016

Mesenchymal stem cells (MSCs) are widely used in cartilage tissue engineering to repair articular cartilage defects. However, hypertrophy of chondrocytes derived from MSCs might hinder the stabilization of hyaline cartilage. Thus, it is very important to find a suitable way to maintain the chondrogenic phenotype of chondrocytes. It has been reported that cordycepin has anti-inflammatory and anti-tumor functions. However, the role of cordycepin in chondrocyte hypertrophy remains unclear. Therefore, the objective of this study was to determine the effect of cordycepin on chondrogenesis and chondrocyte hypertrophy in MSCs and ATDC5 cells. Cordycepin upregulated chondrogenic markers including Sox9 and collagen type II while down-regulated hypertrophic markers including Runx2 and collagen type X. Further exploration showed that cordycepin promoted chondrogenesis through inhibiting Nrf2 while activating BMP signaling. Besides, cordycepin suppressed chondrocyte hypertrophy through PI3K/Bapx1 pathway and Notch signaling. Our results indicated cordycepin had the potential to maintain chondrocyte phenotype and reconstruct engineered cartilage.

• Journal of Periodontal and Implant Science
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• 제42권3호
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• pp.73-80
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• 2012

Purpose: The purpose of this study was to investigate the effects of the immunosuppressants FK506 and cyclosporin A (CsA) on the osteogenic differentiation of rat mesenchymal stem cells (MSCs). Methods: The effect of FK506 and CsA on rat MSCs was assessed in vitro. The MTT assay was used to determine the deleterious effect of immunosuppressants on stem cell proliferation at 1, 3, and 7 days. Alkaline phosphatase (ALP) activity was analyzed on days 3, 7, and 14. Alizarin red S staining was done on day 21 to check mineralization nodule formation. Real-time polymerase chain reaction (RT-PCR) was also performed to detect the expressions of bone tissue-specific genes on days 1 and 7. Results: Cell proliferation was promoted more in the FK506 groups than the control or CsA groups on days 3 and 7. The FK506 groups showed increased ALP activity compared to the other groups during the experimental period. The ALP activity of the CsA groups did not differ from the control group in any of the assessments. Mineralization nodule formation was most prominent in the FK506 groups at 21 days. RT-PCR results of the FK506 groups showed that several bone-related genes-osteopontin, osteonectin, and type I collagen (Col-I)-were expressed more than the control in the beginning, but the intensity of expression decreased over time. Runx2 and Dlx5 gene expression were up-regulated on day 7. The effects of 50 nM CsA on osteonectin and Col-I were similar to those of the FK506 groups, but in the 500 nM CsA group, most of the genes were less expressed compared to the control. Conclusions: These results suggest that FK506 enhances the osteoblastic differentiation of rat MSCs. Therefore, FK506 might have a beneficial effect on bone regeneration when immunosuppressants are needed in xenogenic or allogenic stem cell transplantation to treat bone defects.

• 대한한방내과학회지
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• 제36권4호
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• pp.518-526
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• 2015

Objectives This study investigated the effect of purified safflower (Carthamus tinctorius Linne) and safflower seed (Carthamus tinctorius L. seed; CS) extract, using hot water and ethanol extract methods , on the osteogenic differentiation of MC3T3E1 cells.Methods The safflower and safflower seed were extracted with hot water and ethanol. The samples were concentrated by a rotary evaporator and then freeze-dried using a freeze-dryer. The MC3T3E1 cells were propagated and maintained in DMEM (Gibco) containing 10% FBS and a 1% antibiotic antimycotic solution. To induce osteogenic differentiation, the cells were treated for 14 days with DMEM with 10 mM β-glycerophosphate and 50 μM ascorbic acid. Extract doses were confirmed by the results of an MTT assay, and treatment of the extracts was performed in a differentiation medium every two days. The ALP staining and activity were tested after osteogenic differentiation for five days, and after 14 days, osteogenic differentiation was determined by alizarin red S staining. The mRNA expressions of osteogenic-related genes were quantified using quantitative real-time PCR.Results In the results of the MTT assay, all concentrations of safflower extracts had no toxicity in the MC3T3El cells. But in the groups of 100 ng/ml and 200 ng/ml concentrations of safflower seed extracts, the cell viability was significantly reduced by up to 40-50%. So we fixed the treatment concentration of the extract at 50 ng/ml. In the ALP and alizarin red S staining, all extract groups increased osteogenic differentiation compared with the control group. The water-safflower extract group showed the highest mRNA level of Alp, Runx2, and Dlx5 genes. The mRNA level of Ocn, an osteogenic gene related to late-stage differentiation, in the ethanol-safflower extract group increased the mineralization more significantly than in other groups.Conclusions These data suggest that the extract of safflower increases the osteoblastic differentiation activates of MC3T3E1 cells like the extract of safflower seed. The water-extract and ethanol-extract of safflower have effects on different stages of osteogenesis in MC3T3El. Not only safflower seed but also safflower will be useful therapeutic reagents for age-associated chronic diseases such as osteoporosis.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제27권3호
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• pp.214-220
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• 2001

안면비대칭이나 반안면왜소증과 같은 구강악안면영역의 기형에 대한 진단 및 치료 계획 수립을 위하여 두개 악안면 구조물에 대한 총체적인 접근이 필요하다. 이에 두부 규격 방사선 사진을 이용한 3차원 두부 계측 방사선 시스템의 임상적 적용이 계속 시도되었으나 주로 두 방향의 방사선원으로부터 동시에 얻어진 standard Broadbent-Bolton cephalometer에 한정되어 있었다. 이에 본 연구에서는 단일방사선원에 의하여 촬영된 한 쌍의 정모 및 측모두부방사선 사진으로 정확한 3차원 좌표치를 얻을 수 있는 방법을 개발하고 이렇게 얻어진 3차원 좌표치의 정확도와 재현성을 검증하여 다음과 같은 결과를 얻었다. 1. 정모, 측모 두부 방사선 사진을 동시에 촬영하지 않음으로 인하여 생기는 오차를 보정해주는 수학적인 공식을 산출하고 이 공식에 의거하여 정모 두부 방사선 사진에 기준한 측모 두부 방사선 계측 사진의 위치 결정시의 오차를 건조 두개골을 이용하여 검증한 결과 1차와 2차 측정시 각각 평균 $0.46{\pm}1.21^{\circ}, $0.33{\pm}0.90^{\circ}로 나타났다. 2. 단일 방사선원으로부터 3차원 계측점의 좌표값을 얻을때의 재현성을 파악하기 위한 임상적 계측 결과 1, 2차측정간의 오차가 절대값 평균 0.54mm ($-2.99{\sim}2.26mm$)로 나타났으며 이때 계측점 인식의 오차는 평균 $1.2{\pm}1.6mm$ 였다. 위와 같은 결과를 종합하여 볼 때 정모 두부 방사선 사진에 의거한 측모 두부 방사선 계측사진의 위치를 결정한 후 두부위치 보정공식을 도입하면 임상적으로 충분히 재현성 있는 3차원 계측을 할 수 있다는 것을 알 수 있었다.을 증가시킨다. - PKC pathway는 MAPK pathway를 경유하여 Runx2의 전사 활성을 조절한다.있을 것으로 사료된다.대구치의 원심경사는 필연적으로 일어나며, 이를 최소화하기 위한 노력이 계속되어야 할 것으로 사료된다.글래스 아이오노머 시멘트 사이에는 유의한 차이가 없었다.착제의 종류에 따른 전단결합강도를 비교한 결과, 영구치와 유치 모두에서 Clearfil SE Bond를 사용한 군의 전단결합강도가 가장 높았으며 AQ Bond를 사용한 군의 전단결합강도가 가장 낮았다.본에서 III군에 비해 크기가 큰 기포가 더욱 많이 관찰되었다.e의 약동학은 cimetidine에 의해 유의한 차이를 보였으며 CYP1A2유전자형에 따른 영향은 관찰할 수 없었다. CYP1A2유전자형에 따른 생체내 대사능을 관찰하는 실험이 향후 이루어 져야 할 것으로 사료된다.san film보다 큰 수증기 투과도를 보였다.적으로 유의한 차이를 보이지 않았다.y tissue layer thinning은 3 군모두에서 관찰되었고 항암 3 일군이 가장 심하게 나타났다. 이상의 실험결과를 보면 술전 항암제투여가 초기에 시행한 경우에는 조직의 치유에 초기 5 일정도까지는 영향을 미치나 7 일이 지나면 정상범주로 회복함을 알수 있었고 실험결과 항암제 투여후 3 일째 피판 형성한 군에서 피판치유가 늦어진 것으로 관찰되어 인체에서 항암 투여후 수술시기는 인체면역계가 회복하는 시기를 3주이상 경과후 적어도 4주째 수술시기를 정하는 것이 유리하리라 생각되었다.한 복합레진은 개발의 초기단계이며, 물성의 증가를 위한 연구가 필요할 것으로 사료된다.또 다른 약물인 glycyrrhetinic acid($100{\mu}M$)도 CCh 자극으로 인한 타액분비를 억제하였다. 이상의 결과로 미루어

• Maxillofacial Plastic and Reconstructive Surgery
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• 제29권4호
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• pp.279-288
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• 2007

In the present study, we focused on stem cells in the dental papilla of the tooth germ. The tooth germ, sometimes called the tooth bud, is the primordial structure from which a tooth is formed. The tooth germ consists of the enamel organ, the dental papilla, and the dental follicle. The dental papilla lies below a cellular aggregation of the enamel organ. Mesenchymal cells within the dental papilla are responsible for formation of dentin and pulp of a tooth. Tooth germ disappears as a tooth is formed, but that of a third molar stays in the jawbone of a human until the age of 10 to 16, because third molars grow slowly. Impacted third molar tooth germs from young adults are sometimes extracted for orthodontic treatment. In the present study, we evaluated the osteogenic activity and mineralization of cultured human dental papilla-derived cells. Dental papillas were harvested from mandible during surgical extraction of lower impacted third molar from 3 patients aged 13-15 years. After passage 3, the dental papilla-derived cells were trypsinized and subsequently suspended in the osteogenic induction DMEM medium supplemented with 10% fetal bovine serum, 50 g/ml L-ascorbic acid 2-phosphate, 10 nM dexamethasone and 10 mM -glycerophosphate at a density of $1\;{\times}10^6\;cells/dish$ in a 100-mm culture dish. The dental papilla-derived cells were then cultured for 6 weeks and the medium was changes every 3 days during the incubation period. Dental papilla-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 7 of culture period, then decreased in intensity during the culture period. ALP mRNA level was largely elevated at 1 weeks and gradually decreased with culture time. Osteocalcin mRNA expression appeared at day 14 in culture, after that its expression continuously increased in a time-dependent manner up to day 28. The expression remained constant thereafter. Runx2 expression appeared at day 7 with no detection thereafter. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. Osteocalcin secretion was detectable in the culture medium from 1 week. The secretion of osteocalcin from dental papilla-derived cells into the medium greatly increased after 3 weeks although it showed a shallow increase by then. In conclusion, our study showed that cultured human dental papilla-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix.