• 대한치과교정학회지
• /
• 제34권6호
• /
• pp.514-525
• /
• 2004

흡연이 골형성 세포의 기능을 억제한다는 것은 이미 잘 알려져 있으나 대다수의 연구가 주로 역학조사에 기초하여 이루어져 왔다. 본 연구의 목적은 흠연과 골형성 세포의 상관관계를 실험적으로 평가하기 위한 것이다. 본 연구를 위해 인간 골육종 세포인 MG63 조골제포주를 이용하였으며, 니코틴이 조골세포의 증식, 염기성 인산분해효소의 활성, 오스테오칼신곽 오스테오프로테제린의 생성 그리고 mRNA발현에 미치는 영향을 관찰하였다 MG63조골세포를 니코틴 농도가 각각 $1.0{\mu}M$,\;1.0mM, 2.5mM, 5.0mM, 7.5mM, 그리고 10.0mM,이 함유된 배지에서 1일, 2일, 3일 그리고 6일 동안 배양 실험을 하여 다음과 같은 결과를 얻었다. MG63 조골세포의 증식이 저농도의 니코틴에서는 일시적으로 활성화되었지만 5.0mM 이상의 고농도에서는 억제되었다. 염기성 인산분해효소의 활성은 니코틴 농도가 증가함에 따라 감소하였다. 오스테오칼신 생성양은, 배양 1일 후, 5.0mM 이하의 농도에서 증가하였으나 7.5mM 이상의 고농도에서는 감소하였다. MG63 조골세포를 3일 배양한 경우, 오스테오칼신 생성양은 1.0mM의 저농도에서도 감소하였다 니코틴이 오스테오칼신 단백질 생성과 오스테오칼신 mRNA생성에 미치는 영향은 1일과 3일에서 다소 차이가 있었다. 오스테오프로테제린의 생성양은 모든 실험군에서 니코틴을 함유하지 않은 대조군보다 감소하였다. 하지만 mRNA 수치는 단백질 생성과 상반되게 7.5mM (3일), 5.0mM (6일) 이상의 고농도에서 증가하였다.

• Journal of Periodontal and Implant Science
• /
• 제40권1호
• /
• pp.39-44
• /
• 2010

Purpose: Bone tissues for clinical application can be improved by studies on osteoblast differentiation. Runx2 is known to be an important transcription factor for osteoblast differentiation. However, bone morphogenetic protein (BMP)-2 treatment to stimulate Runx2 is not sufficient to acquire enough bone formation in osteoblasts. Therefore, it is necessary to find other regulatory factors which can improve the transcriptional activity of Runx2. The erythroblast transformation-specific (ETS) transcription factor family is reported to be involved in various aspects of cellular proliferation and differentiation. Methods: We have noticed that the promoters of osteoblast differentiation markers such as alkaline phosphatase (Alp), osteopontin (Opn), and osteocalcin (Oc) contain Ets binding sequences which are also close to Runx2 binding elements. Luciferase assays were performed to measure the promoter activities of these osteoblast differentiation markers after the transfection of Runx2, myeloid Elf-1-like factor (MEF), and Runxs+MEF. Reverse-transcription polymerase chain reaction was also done to check the mRNA levels of Opn after Runx2 and MEF transfection into rat osteoblast (ROS) cells. Results: We have found that MEF, an Ets transcription factor, increased the transcriptional activities of Alp, Opn, and Oc. The addition of Runx2 resulted in the 2- to 6-fold increase of the activities. This means that these two transcription factors have a synergistic effect on the osteoblast differentiation markers. Furthermore, early introduction of these two Runx2 and MEF factors significantly elevated the expression of the Opn mRNA levels in ROS cells. We also showed that Runx2 and MEF proteins physically interact with each other. Conclusions: Runx2 interacts with MEF proteins and binds to the promoters of the osteoblast markers such as Opn nearby MEF to increase its transcriptional activity. Our results also imply that osteoblast differentiation and bone formation can be increased by activating MEF to elicit the synergistic effect of Runx2 and MEF.

• 대한한의학회지
• /
• 제24권3호
• /
• pp.35-44
• /
• 2003

Objective : This study was performed to evaluate the effects of Palmijihwang-hwan (Baweidehuang-wan) and Obaeja (Galla Rhois) on the regeneration of periodontal tissue. Methods : In this study, we used MC3T3-El cells, such as osteoblast-like cell lines and human periodontal ligament fibroblasts, for experimental material. We separated each type of cells into a control group and an experimental group. In the control group, the cells were cultivated for 48 hours with distilled water and media which contained 10% fetal bovine serum (FBS) and penicillin (l00unit/ml)-streptomycin ($l00{\mu\textrm{g}}/ml$) at $37^{\circ}$ in 5% $CO_2$ gas. In the experimental group, the cells were cultivated for 48 hours with Palmijihwang-hwan extract and Obaeja extract (concentrations $1{\mu\textrm{g}}/ml,{\;}25{\mu\textrm{g}}/ml,{\;}50{\mu\textrm{g}}/ml$) under the same conditions as the control group. Investigating the regeneration of periodontal tissue was performed by evaluating proliferation, the activity of alkaline phosphatase and the synthetic ability of proteins using those cultivated cells by means of microculture tetrazolium (MTT) assay, alkaline phosphatase substrate kit and protein assay kit. Results : 1. In vitro, Palmijihwang-hwan extract increased the proliferation of MC3T3-El cells. 2. In vitro, Obaeja extract increased the activity of alkaline phosphatase and the synthetic ability of protein in MC3T3-El cells and human periodontal ligament fibroblasts depending on Obaeja extract's concentration. Conclusion : Obaeja extract can be developed as a subsidiary medicine for the regeneration of periodontal tissue. Further studies to evaluate the different concentrations the Obaeja extract and clinical trials in vivo are suggested.

• International Journal of Oral Biology
• /
• 제33권3호
• /
• pp.105-111
• /
• 2008

Thrombospondins (TSP-1, TSP-2) are secretory extracellular glycoproteins that are involved in a variety of physiological processes such as tumor cell adhesion, invasion, and metastasis. The present study was undertaken to elucidate the involvement of thrombospondins in the adhesion of osteoblast-like cells using the TSP-1 or TSP-2 antisense MG63 and MC3T3-E1 cell lines. For downregulation of TSPs expression, we prepared antisense constructs for TSP-1 and TSP-2 using the pREP4 an episomal mammalian expression vector, which be able to produce the specific antisense oligonucleotides around chromosome. MG63 and MC3T3-E1 osteoblast-like cells were transfected with the antisense constructs and nonliposomal Fugene 6, and then selected under hygromycin B (50 ${\mu}g/ml$) treatment for 2 weeks. Western blot analysis revealed that expression of the TSP proteins was downregulated in the antisense cell lines. The cell adhesion assay showed that adhesive properties of TSP-1 and TSP-2 antisense MG63 cells on the polystyrene culture plate were reduced to 17% and 21% of the control cells, respectively, and those of the TSP-1 and TSP-2 antisense MC3T3-E1 cells also decreased to 19% and 27% of control, respectively. Adhesion of TSP-1 and TSP-2 antisense MC3T3-E1 cells on Type I collagen-coated culture plate decreased to 27% and 76%, respectively. These results indicate that TSP-1 and TSP-2 proteins may have an important role in adhesion of osteoblast-like cells to extracellular matrix.

• 한국재료학회지
• /
• 제20권7호
• /
• pp.392-400
• /
• 2010

The electrospinning process was established as a promising method to fabricate nano and micro-textured scaffolds for tissue engineering applications. A BCP-loaded PCL micro-textured scaffold thus can be a viable option. The biocompatibility as well as the mechanical properties of such scaffold materials should be optimized for this purpose. In this study, a composite scaffold of poly ($\varepsilon$-caprolactone) (PCL)-biphase calcium phosphate (BCP) was successfully fabricated by electrospinning. EDS and XRD data show successful loading of BCP nano particles in the PCL fibers. Morphological characterization of fibers shows that with a higher loaded BCP content the fiber surface was rougher and the diameter was approximately 1 to 7 ${\mu}m$. Tensile modulus and ultimate tensile stress reached their highest values in the PCL- 10 wt% BCP composite. When content of nano ceramic particles was low, they were dispersed in the fibers as reinforcements for the polymer matrix. However, at a high content of ceramic particles, the particles tend to agglomerate and lead to decreasing tensile modulus and ultimate stress of the PCL-BCP composite mats. Therefore, the use of nano BCP content for distribution in fiber polymer using BCP for reinforcement is limited. Tensile strain decreased with increasing content of BCP loading. From in vitro study using MG-63 osteoblast cells and L-929 fibroblast like cells, it was confirmed that electrospun PCL-BCP composite mats were biocompatible and that spreading behavior was good. As BCP content increased, the area of cell spreading on the surface of the mats also increased. Cells showed the best adherence on the surface of composite mats at 50 wt% BCP for both L-929 fibroblast-like cells and MG-63 osteoblast cell. PCL- BCP composites are a promising material for application in bone scaffolds.

• 한국약용작물학회:학술대회논문집
• /
• 한국약용작물학회 2008년도 춘계학술발표회
• /
• pp.182-183
• /
• 2008

• 대한치주과학회:학술대회논문집
• /
• 대한치주과학회 1999년도 제39회 종합학술대회 연제초록
• /
• pp.89-90
• /
• 1999

• Carbon letters
• /
• 제22권
• /
• pp.42-47
• /
• 2017

In this paper, the in vitro biocompatibility of graphene film (GF) with osteoblasts was evaluated through cell adhesion, viability, alkaline phosphatase activity, F-actin and vinculin expressions, versus graphite paper as a reference material. The results showed that MG-63 cells exhibited stronger cell adhesion, better proliferation and viability on GF, and osteoblasts cultured on GF exhibited vinculin expression throughout the cell body. The rougher and wrinkled surface morphology, higher elastic modulus and easy out-of-plane deformation associated with GF were considered to promote cell adhesion. Also, the biomineralization of GF was assessed by soaking in simulated body fluid, and the GF exhibited enhanced mineralization ability in terms of mineral deposition, which almost pervaded the entire GF surface. Our results suggest that graphene promotes cell adhesion, activity and the formation of bone-like apatite. This research is expected to facilitate a better understanding of graphene-cell interactions and potential applications of graphene as a promising toughening nanofiller in bioceramics used in load-bearing implants.

• Journal of Periodontal and Implant Science
• /
• 제41권6호
• /
• pp.293-301
• /
• 2011

Purpose: We have previously reported that tetra-cell adhesion molecule (T-CAM) markedly enhanced the differentiation of osteoblast-like cells grown on anorganic bone mineral (ABM). T-CAM comprises recombinant peptides containing the Arg- Gly-Asp (RGD) sequence in the tenth type III domain, Pro-His-Ser-Arg-Asn (PHSRN) sequence in the ninth type III domain of fibronectin (FN), and the Glu-Pro-Asp-Ilu-Met (EPDIM) and Tyr-His (YH) sequence in the fourth fas-1 domain of ${\beta}$ig-h3. Therefore, the purpose of this study was to evaluate the cellular activity of osteoblast-like cells and the new bone formation on ABM coated with T-CAM, while comparing the results with those of synthetic cell binding peptide (PepGen P-15). Methods: To analyze the cell viability, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed, andto analyze gene expression, northernblot was performed. Mineral nodule formations were evaluated using alizarin red stain. The new bone formations of each group were evaluated using histologic observation and histomorphometrc analysis. Results: Expression of alkaline phosphatase mRNA was similar in all groups on days 10 and 20. The highest expression of osteopontin mRNA was observed in the group cultured with ABM/P-15, followed by those with ABM/T-CAM and ABM on days 20 and 30. Little difference was seen in the level of expression of collagen type I mRNA on the ABM, ABM/T-CAM, and ABM/P-15 cultured on day 20. There were similar growth and proliferation patterns for the ABM/T-CAM and ABM/P-15. The halo of red stain consistent with $Ca^{2+}$ deposition was wider and denser around ABM/T-CAM and ABM/P-15 particles than around the ABM particles. The ABM/T-CAM group seemed to have bone forming bioactivity similar to that of ABM/P-15. A complete bony bridge was seen in two thirds of the defects in the ABM/T-CAM and ABM/P-15 groups. Conclusions: ABM/T-CAM, which seemed to have bone forming bioactivity similar to ABM/P-15, was considered to serve as effective tissue-engineered bone graft material.

• 대한치과보철학회지
• /
• 제41권3호
• /
• pp.300-318
• /
• 2003

Statement of problem: The success of implants depends on intimate and direct contact of implant material on bone tissue and on functional relationship with soft tissue contact. Creation and maintenance of osseointegration depend on the understanding of the tissue's healing, repairing, and remodeling capacity and these capacities rely on cellular behavior. Altering the surface properties can modify cellular responses such as cell adhesion, cell motility, bone deposition, Therefore, various implant surface treatment methods are being developed for the improved bone cell responses. Purpose: The purpose of this study was to evaluate the responses of osteoblast-like cells to surface-modified titanium. Materials and Methods: The experiment was composed of four groups. Group 1 represented the electropolished surface. Group 2 surfaces were machined surface. Group 3 and Group 4 were anodized surfaces. Group 3 had low roughness and Group 4 had high roughness. Physicochemical properties and microstructures of the discs were examined and the responses of osteoblast-like cells to the discs were investigated. The microtopography was observed by SEM. The roughness was measured by three-dimension roughness measuring system. The microstructure was analyzed by XRD, AES. To evaluate cell responses to modified titanium surfaces, osteoblasts isolated from calvaria of neonatal rat were cultured. Cell count, morphology, total protein measurement and alkaline phosphatase activities of the cultures were examined. Results and Conclusion: The results were as follows 1. The four groups showed specific microtopography respectively. Anodized group showed grain structure with micropores. 2. Surface roughness values were, from the lowest to the highest, electropolished group, machined group, low roughness anodized group, and high roughness anodized group. 3. Highly roughened anodized group was found to have increased surface oxide thickness and surface crystallinity. 4. The morphology of cells, flattened or spherical, were different from each other. In the electropolished group and machined group, the cells were almost flattened. In two anodized groups, some cells were spherical and other cells were flattened. And the 14 day culture cells of all of the groups were nearly flattened due to confluency. 5. The number of attached cells was highest in low roughness anodized group. And the machined group had significantly lower cell count than any other groups(P<.05). 6. Total protein contents showed no difference among groups. 7. The level of alkaline phosphatase activities was higher in the anodized groups than electropolished and machined groups(P<.05).