• Journal of Periodontal and Implant Science
• /
• 제30권1호
• /
• pp.27-39
• /
• 2000

To evaluate the effects of Dexamethasone(Dex), Platelet derived growth factor-BB(PDGF) and combination of Dex and PDGF(DP) on the growth and differentiation of MC3T3-E1 cells, Dex($10^{-7}\;M$) and PDGF(10 ng/ml) in experimental group were added to the cells at the days 5, 10, 15, 20, 25 and examined for cell proliferation activities, DNA synthesis activities, ALP activities and bone nodule formation. The results were as follows : 1. In Dex group, cell proliferation, DNA synthesis and ALP activities were lower until 15 days when compared to the control group. Bone nodules formation were shown at 10 days. 2. In PDGF group, cell proliferation and DNA synthesis activities were higher until 15 days and ALP activities were lower when compared to the control and Dex groups. Bone nodules formation were shown at 20 days. 3. In DP group, cell proliferation and DNA synthesis activities of PDGF were suppressed by Dex and synergistic effects of combination of Dex and PDGF on ALP activities were shown at days 5 when compared to control and Dex groups. Bone nodules formation activities of Dex were suppressed by PDGF.

• 한국해양바이오학회지
• /
• 제15권1호
• /
• pp.24-32
• /
• 2023

The Laminaria japonica Aresch (Sea tangle) belongs to the brown algae and has a long history as a food material in Asia, including Korea. Recent studies have found that the fermented Sea tangle extract (FST) inhibited the differentiation of osteoclasts and protected osteoblasts from oxidative damage. This study aims to explore the possibility that FST can induce the differentiation of osteoblasts and identify the responsible mechanism. According to our results, FST induced differentiation into osteogenic cells in the presence of osteoblastic MC3T3-E1 cells under non-toxic conditions.. This finding was confirmed by phalloidin staining, increased alkaline phosphatase activity, and calcium deposition. Additionally, it was found that this process was achieved by increasing the expression of key factors involved in osteoblast differentiation, such as runt-related transcription factor-2, osterix, β-catenin, and bone morphogenetic protein-2. Moreover, FST increased autophagy, which may contribute to the maintenance of the bone formation homeostasis, and is associated with the activation of the phosphatidylinositol 3-kinase/Akt and mitogen-activated protein kinase signaling pathways. Although further research about the bioactive substances contained in FST and the tests of their efficacy are required, the results of this study indicate that FST has incredible applicability as a functional material for maintaining the bone homeostasis.

• 농약과학회지
• /
• 제6권2호
• /
• pp.96-104
• /
• 2002

Paraquat의 세포독성을 알아보기 위하여 NIH 3T3 섬유모세포에 적용한 후 MTT와 NR 분석법을 이용하여 세포독성을 측정하고, paraquat의 세포독성에 대한 3-MC의 독성경감효과를 알아보기 위하여 Spraque Dawley계 수컷 랫드에 paraquat 단독 및 paraquat와 3-MC 병용투여 후 랫드의 폐를 경시적으로 채취하여 관찰하였다. Paraquat의 NIH 3T3 섬유모세포에 대한 $MTT_{50}$은 $1668.97{\mu}M$, $NR_{50}$은 $1030.25{\mu}M$로 산출되어 Borefreund와 Puemer(1984)의 독성판정기준에 의하면 저독성 물질이었다. Paraquat 단독 투여 군은 H&E 염색에서 3시간째부터 폐 모세혈관 내에 적혈구 수가 증가하기 시작하여 24시간째에는 충혈상태에 이르렀으며, 폐포사이 중격에서는 큰폐포상피수가 증가하였다. 또한 폐 조직을 둘러싸고 있는 결합조직 내에는 임파구, 대식세포 및 다형핵 백혈구 등이 다수 관찰되었고, 48시간째부터 폐포사이 중격과 폐포내에 폐포큰포식세포가 증가하기 시작하여 96시간째에는 다수의 폐포큰포식세포가 관찰되었다. Verhoeff의 iron hematoxylin 염색에서도 paraquat 단독 투여 후 24시간째에 조직변화가 가장 심하였고, 교원섬유량의 급격한 증가, 폐포의 넓이와 폐포 구멍(alveolar pore) 간격의 확장 등이 관찰되었다. 한편, paraquat와 3-MC 병용투여군은 paraquat 단독 투여 군에 비하여 조직변화가 약하게 관찰되었는데, 병용투여 후 3시간째에는 단독투여 3시간째의 소견과 유사하였으나 점차 회복되어 폐 모세혈관 내에 적혈구 수가 증가하여 24시간째에는 대조군의 구조와 거의 유사하였다. 또한 폐 조직을 둘러싸고 있는 결합조직과 임파소절에서도 paraquat 단독 투여 군에서 보였던 변화가 거의 관찰되지 않았다. Verhoeff의 iron hematoxylin 염색에서도 병용투여 후 24시간째에는 교원섬유량이 단독 투여 군에 비하여 크게 감소하였고 폐포와 폐포 구멍의 넓이도 대조군과 유사하였다.

• 대한치과교정학회지
• /
• 제27권6호
• /
• pp.929-941
• /
• 1997

가장 바람직한 교정력은 환자에게 불편감을 주지 않고 치조골 상실과 치근흡수와 같은 조직의 손상없이 가장 빨리 치아를 이동시키는 힘이다. 최적의 교정력을 얻기 위하여 그 동안 많은 방법들이 시도되어 왔으며 최근에는 자석의 사용이 고려되고 있다. 본 연구는 Sm-Co 자석의 정자기장이 효소와 세포 활성에 미치는 영향을 알아보기 위하여 시행되었다. 적혈구 침강속도가 측정되었으며, 철이온과 관련된 효소 (Catalase, NO synthase)와 철이온과 무관한 효소 (Lactic dehydrogenase)의 활성과 세포내 합성은 Spectrophotometer를 이용하여 측정되었으며, 조골세포 $MC_{3}T_3-E_1$의 성장과 증식은 Crystal violet 염색법과 ${^3}H$-thymidine incorporation에 의한 DNA합성능을 측정하였다. 실험군의 적혈구는 표면자기장이 1,400 G (gauss)인 자석에, 효소와 조골세포는 7,000 G의 정자기장에 노출시키고, 정자기장에 노출시키지 않은 경우와 비교하여 다음과 같은 결과를 얻었다. 1. 적혈구 침강속도는 정자기장의 영향을 받지 않았다. 2. Catalase와 Lactic dehydrogenase의 활성은 정자기장의 영향을 받지 않았다. 3. NO synthase와 Lactic dehydrogenase의 세포내 합성은 정자기장의 영향을 받지 않았다. 4. 세포배양된 조골세포 $MC_{3}T_3-E_1$의 성장과 증식은 정자기장의 영향을 받지 않았다. 이상의 결과로 보아 정자기장은 효소와 세포 활성에 대한 영향이 없는 것으로 사료되었다.

• Journal of Periodontal and Implant Science
• /
• 제29권4호
• /
• pp.751-765
• /
• 1999

Several growth factors and polypeptidesare not commonly yet used for regenerators of bone tissue or alveolar bone because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many natural medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, anti-inflammatory and regenerative potential of periodontal tissues. Olibanum, Myrrha, Phlomis Radix, and Cimicifugae Rhizoma have been traditionally used as a drug for treatment of bone disease in oriental medicine. The objective of this study was to examine the ability of alkaline phosphatase(ALP) synthesis of rat calvarial osteoblast(MC3T3-E1) when several natural medicines were supplemented. MC3T3-E1 cells were cultured with ${\alpha}$-MEM(negative control), dexamethasone(positive control), and each natural medicines for 3 and 5 days. And then ALP synthesis was measured by spectrophotometer for enzyme activity and by naphthol AS-BI staining for morphometry. All of the natural medicines induced higher activity of ALP synthesis than the negative controls. Especially Olibanumind uced the higher activity than the positive controls (p<0.05). In the aspects of culturing time, except Cimicifugae Rhizoma, the natural medicines induced higher activity of ALP synthesis at 5 days than at 3 days (p<0.05). In morphometry, all of the natural medicines showed statistical significance compared to the negative control (p<0.05). Myrrha a n d Phlomis Radix showed larger positively stained area at 5days than at 3 days, whereas the others did not showed the difference between at 5 and at 3 days(p<0.05). These results indicate that several natural medicines have an inducing ability of ALP synthesis in MC3T3-E1 cells.

• Nutritional Sciences
• /
• 제9권4호
• /
• pp.233-239
• /
• 2006

Porous matrices of bioactive polymers such as polyglycolic acid (PGA) or polylactic acid (PLA) can be used as scaffolds in bone tissue growth during bone repair process. These polymers are highly porous and serve as a template for the growth and organization of new bone tissues. We evaluated the effect of PGA and PLA polymers on osteoblastic MC3T3-E1 cell extracellular mineralization. MC3T3-E1 cells were cultured in a time-dependent manner -1, 15, 25d as appropriate - for the period of bone formation stages in one of the five culture circumstances, such as normal osteogenic differentiation medium, PGA-plated, fetal bovine serum (FBS)-plated, PGA/FBS-coplated, and PLA-plated For the evaluation of bone formation, minerals (Ca, Mg, Mn) and alkaline phosphatase activity, a marker for osteoblast differentiation, were measured Alizarin Red staining was used for the measurement of extracellular matrix Ca deposit During the culture period, PGA-plated one was reabsorbed into the medium more easily and faster than the PLA-plated one. At day 15, at the middle stage of bone formation, cellular Ca and Mg levels showed higher tendency in PGA- or PLA-plated treatments compared to non-plated control and at day 25, at the early late stage of bone formation, all three cellular Ca, Mg or Mn levels showed higher tendency as in order of PGA-related treatments and PLA-plated treatments, compared to control even without significance. Medium Ca, Mg or Mn levels didn't show any consistent tendency. Cellular ALP activity was higher in the PGA- or PLA-plated treatments compare to normal osteogenic medium treatment PGA-plated and PGA/FBS-plated treatments showed better Ca deposits than other treatments by measurement of Alizarin Red staining, although PLA-plated treatment also showed reasonable Ca deposit. The results of the present study suggest that biodegradable material, PGA and also with less extent for PLA, can be used as a biomaterial for better extracellular matrix mineralization in osteoblastic MC3T3-E1 cells.

• Journal of Nutrition and Health
• /
• 제51권5호
• /
• pp.379-385
• /
• 2018

Purpose: Zinc (Zn) is an essential trace element for bone mineralization and osteoblast function. We examined the effects of Zn deficiency on osteoblast differentiation and mineralization in MC3T3-E1 cells. Methods: Osteoblastic MC3T3-E1 cells were cultured at concentration of 1 to $15{\mu}M$ $ZnCl_2$ (Zn- or Zn+) for 5, 15 and 25 days up to the calcification period. Extracellular matrix mineralization was detected by staining Ca and P deposits using Alizarin Red and von Kossa stain respectively, and alkaline phosphatase (ALP) activity was detected by ALP staining and colorimetric method. Results: Extracellular matrix mineralization was decreased in Zn deficiency over 5, 15, and 25 days. Similarly, staining of ALP activity as the sign of an osteoblast differentiation, was also decreased by Zn deficiency over the same period. Interestingly, the gene expression of bone-related markers (ALP, PTHR; parathyroid hormone receptor, OPN; osteopontin, OC; osteocalcin and COLI; collagen type I), and bone-specific transcription factor Runx2 were downregulated by Zn deficiency for 5 or 15 days, however, this was restored at 25 days. Conclusion: Our data suggests that Zn deficiency inhibits osteoblast differentiation by retarding bone marker gene expression and also inhibits bone mineralization by decreasing Ca/P deposition as well as ALP activity.

• 대한치과보철학회지
• /
• 제46권3호
• /
• pp.227-237
• /
• 2008

STATEMENT OF PROBLEM: Zirconium oxide can be a substitute to titanium as implant materials to solve the esthetic problems of dark color in the gingival portion of implant restorations. PURPOSE: This study was performed to define attachment and growth behavior of osteoblast- like cells cultured on grooved surfaces of zirconium oxide and evaluate the genetic effect of zirconium oxide surfaces using the reverse transcriptase-polymerase chain reaction (RT-PCR). MATERIAL AND METHODS: MC3T3-E1 cells were cultured on (1) commercially pure titanium discs with smooth surface (T group), (2) yttrium-stabilized tetragonal zirconia polycrystal (Y-TZP) with machined surface (ZS group), and (3) Y-TZP with $100{\mu}m$ grooves (ZG group). Cell proliferation activity was evaluated through MTT assay and cell morphology was examined by SEM. The mRNA expression of Runx2, alkaline phosphatase, osteocalcin, TGF-${\beta}1$, IGF-1, G3PDH in E1 cells were evaluated by RT-PCR. RESULTS: From the MTT assay, after 48 hours of adhesion of MC3T3-E1 cells, the mean optical density value of T group and ZG group significantly increased compared to the ZS group. SEM images of osteoblast-like cells showed that significantly more cells were observed to attach to the grooves and appeared to follow the direction of the grooves. After 24 hours of cell adhesion, more spreading and flattening of cells with active filopodia formation occurred. Results of RT-PCR suggest that T group, ZS group, and ZG group showed comparable osteoblast-specific gene expression after 24 hours of cell incubation. CONCLUSION: Surface topography and material of implants can play an important role in expression of osteoblast phenotype markers. Zirconia ceramic showed comparable biological responses of osteoblast-like cells with titanium during a short-time cell culture period. Also, grooves influence cell spreading and guide the cells to be aligned within surface grooves.

• 대한치과교정학회지
• /
• 제26권4호
• /
• pp.449-454
• /
• 1996

교정력에 의한 치아이동은 세포반응의 결과로 나타난다. 이에 힘의 양상에 따른 세포의 반응을 알아보고자 MC3T3-E1 세포를 24well 배양접시에 배양한 후 밀생상태가 되었을 때, incubator 속의 특수 제작된 사각상자에 배양 접시를 넣고 타이머에 연결된 Diaphram pump를 이용하여 $300gm/cm^2$의 압력을 10분간 가압후 10분간 가압이 중지되도록 한 간헐적 가압군과 지속적으로 가한 가압군으로 하여 각각 실험 24시간, 48시간, 72시간후의 alkaline phosphatase활성도를 대조군과 비교하여 다음과 같은 결과를 얻었다. 1. 가압 24시간 후에서는 각 군간에 alkaline phosphatase 활성도에 유의한 차이가 나타나지 않았다. 2. 가압 48시간 후에서는 간헐적 가압군과 지속적 가압군 모두 대조군 보다 alkaline phosphatase 활성도가 증가 되었다. 3. 가압 72시간 후에서는 간헐적 가압군의 alkaline phosphatase 활성도 증가가 대조군에 비하여 더욱 현저하였으나 지속적 가압군에는 대조군에 비하여 유의한 차이가 나타나지 않았다. 4. 가압 72시간 후에서는 간헐적인 가압군이 지속적인 가압군에 비하여 alkaline phosphatase의 활성도가 증가되었다. 5. 가암에 의한 세포의 뚜렷한 형태학적인 변화는 관찰되지 않았다.

• Imaging Science in Dentistry
• /
• 제35권4호
• /
• pp.191-198
• /
• 2005

Purpose: To characterize the effects of 2-deoxy-D-glucose (2DG) and quercetin (QCT) on cytokine secretion of IL-6, $TGF-\beta$ and gene expression of Col I in irradiated MC3T3-E1 cells Materials and Methods: The MC3T3-El cells were cultured in an a-MEM supplemented with 5mM 2DG or 10mM QCT and then the cells were incubated 12h before irradiation with 2, 4, 6, and 8Gy X-ray using a linear accelerator delivered at a dose rate of 1.5Gy/min. Level of IL-6 and $TGF-\beta$ was determined by ELISA. Also expression of Col I was examined by RT-PCR. Results: In accordance with the radiation dose, the amount of $TGF-\beta$ was not different in RA + QCT, but it showed a peak value in control and RA + 2DG at 4Gy on the 3rd day. However, all groups showed a decreasing tendency dose-dependently in RA+QCT on the 7th day (p<0.01). In accordance with the radiation dose, the amount of IL-6 increased dose-dependently in all groups on the 3rd day. On the 7th and 21st day, all groups showed peak values at 4Gy. RA+QCT showed a slightly increased amount of IL-6 at 2Gy, but it showed a slightly decreased amount at 4, 6, and 8Gy. In accordance with the period of culture after irradiation, the expression of Col I increased dose-dependently in RA+QCT. Conclusion: The result showed that QCT acted as radiosensitizer in the secretion of $TGF-\beta$ and gene expression of Col I during differentiation in irradiated MC3T3-E1 cells at the cellular level.