• Maxillofacial Plastic and Reconstructive Surgery
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• v.38
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• pp.17.1-17.6
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• 2016

Background: The aim of this study is to verify the feasibility of using silk fibroin (SF) as a potential membrane for guided bone regeneration (GBR). Methods: Various cellular responses (i.e., cell attachment, viability, and proliferation) of osteoblast-like MG63 cells cultured on an SF membrane were quantified. After culturing on an SF membrane for 1, 5, and 7 days, the attachment and surface morphology of MG63 cells were examined by optical and scanning electron microscopy (SEM), cell viability was determined using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and cell proliferation was quantified using 4',6-diamidino-2-phenylindole (DAPI) fluorescence staining. Results: Optical microscopy revealed that MG63 cells cultured on the SF membrane proliferated over the 7-day observation period. The viability of cells cultured on SF membranes (SF group) and on control surfaces (control group) increased over time (P < 0.05); however, at respective time points, cell viability was not significantly different between the two groups (P > 0.05). In contrast, cell proliferation was significantly higher in the SF membrane group than in the control group at 7 days (P < 0.05). Conclusions: These results suggest that silk fibroin is a biocompatible material that could be used as a suitable alternative barrier membrane for GBR.

• The Journal of Korean Academy of Prosthodontics
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• v.44 no.5
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• pp.574-583
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• 2006

Statement of problem : The use of permanent magnetics is increasing in implant dentistry. Purpose : This study is to know the effect of permanent magnetics on bone matrix formation of osteoblasts. Materials and methods : The konus abutment-shaped permanent magnetics were connected to the implant fixture, and placed on the culture plate. The osteoblast-like cell Mc3T3E1 were used for cell culture. As the control group, the implants were connected to titanium healing caps, and cultured in the same conditions of experimental group. After 3. 7, 14 days, cells were cultured, and we measured and compared the amount of collagen type I, osteocalcin, which is bone matrix protein by Western immunoblotting analysis. Results: As a result of Western immunoblotting analysis for estimating the amount of bone extracellular matrix, there was no difference between osteoblast of the experimental group and the control group during 3 and 7day-osteoblast culturing. However when cells were cultured for 14days, the amount of bone extracellular matrix was increased, on the experimental group. Conclusion: From these results, magnetic field of permanent magnetics might have effect on bone formation of osteoblast, especially at initial stage of implant placement. Therefore, their clinical application for implant or bone graft could be possible.

• The korean journal of orthodontics
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• v.24 no.1 s.44
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• pp.105-114
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• 1994

The movement of teeth during orthodontic treatment requires bone remodeling process of bone formation and bone resolution. To find out the changes occuring in the cell itself, mechanical stress was applied to the cell populations involved in the bone metabolism. Bone tissue cell populations were isolated from fetal rat calvaria and divided into OC and OB groups. Following results were obtained from measuring the changes in acid & alkaline phosphatease activity, cyclic AMP and $PGE_2$ production in time lapse after the application of mechanical stress. 1. In case of the marker enzyme of specific bone tissue cell, acid phosphatase activity was high in OC group and alkaline phosphatase activity was high in OB group. 2. After the mechanical stress was applied, acid phosphatase activity was decreased in both OC and OB groups and alkaline phosphatase activity was increase in OB group. 3. When the mechanical stress was applied for 15, 30 and 60 minutes, the production of $PGE_2$ increased in both OC and OB groups, as the time span increased. 4. When the mechanical stress was applied for 20 and 40 minutes, the production of $PGE_2$ increased in both OC and OB groups, as the time span increased.

• Journal of Biomedical Engineering Research
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• v.29 no.2
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• pp.159-163
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• 2008

The goal of this study is to investigate the effect and potential of three-dimensional Co-culture of BMSCs (bone marrow stromal Cells) and NP (nucleus pulposus) Cells on the differentiation of BMSCs into NP-like Cells. The NP Cells and BMSCs were isolated and cultured from New Zealand White rabbits. The isolated NP Cells and BMSCs were prepared in different alginate beads. Those two types of beads were separated by a track-etched membrane of $3\;{\mu}m$ pore in a 6-well culture plate. No growth factors were used. In addition to these, NP and BMSC were cultured in the beads independently for control. The number of Cells in Co-culturing system was half of those in two control groups. Proliferation and production of glycosaminoglycan (GAG) were evaluated along with histological observation. The GAG production rate(GAG contents/Cell) of Co-cultured BMSCs were much higher than that of BMSCs cultured alone. The total amounts of GAG produced by BMSCs in Co-culturing system were larger than those produced by BMSCs in control group and were comparable with those produced by NP alone even the number of each Cell was half of BMSCs in Co-culturing system. This study showed the potential of differentiation of BMSCs into NP-like Cells through three-dimensional Co-culture system even without any chemical agents.

• Journal of Periodontal and Implant Science
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• v.27 no.4
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• pp.923-939
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• 1997

This study was performed to evaluate the effect of mixed culture of rat's calvaria cells and periodontal ligament cells on calcification. These cells have been known to do important role on the periodontal tissue regeneration, especially alveolar bone and cementum. Experimental groups were made which based on the different rate of rat's calvaria cells and periodontal ligament cells, and then these cells were cultured with Dulbecco's Modified Eagle's Medium contained with 10% fetal bovine serum, $50{\mu}g/ml$ ascorbic acid, and 10mM/ ml $Na-{\beta}-glycerophosphate$. Each group was characterized by examining the cell proliferation rate, amount of total protein synthesis, alkaline phosphatase activity, and the number of calcified nodules in vitro. In cell proliferation rate , the cells of control groups were cultured Dulbecco's Modified Eagle's Medium contained with 10 % fetal bovine serum. The results were as follows : 1. The cell proliferation rate in control groups decreased stastically significantly along with the decrease of the rate of bone cells at 7 day and 20 day(P < 0.01). 2. The cell proliferation rate in experimental groups decreased stastically significantly along with decrease of the rate of bone cells at 3 day and 14 day(P < 0.01). 3. The amount of total protein synthesis was significantly decreased along with decrease of the rate of bone cells at 3 day and 6 day(p < 0.01). 4. Alkaline phosphatase activity showed reverse time dependent pattern and was significantly decreased along with decrease of the rate of bone cells during the experimental periods (P < 0.01). 5. Calcified nodules were observed in group 1 (Rat calvaria cells alone) for the first time, and the number of calcified nodule decreased stastically significantly along with the decrease of the rate of bone cells at 12 day(P < 0.01). From the above results, When bone cells and periodontal ligament cells were mixed cultured, the cell proliferation rate was mostly dependent on the actual rate of bone cells and same pattern was showed in amount of total protein synthesis, alkalinephosphatase activity, and the number of calcified nodules. And the calcified nodule forming capacity of bone cells was inhibited by periodontal ligament cells

• Journal of Periodontal and Implant Science
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• v.29 no.3
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• pp.483-509
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• 1999

Biodegradable barrier membrane has been demonstrated to have guided bone regeneration capacity on the animal study. The purpose of this study is to evaluate the effects of cultured calvarial cell inoculated on the biodegradable barrier membrane for the regeneration of the artificial bone defect. In this experiment 35 Sprague-Dawley male rats(mean BW 150gm) were used. 30 rats were divided into 3 groups. In group I, defects were covered periosteum without membrane. In group II, defects were repaired using biodegradable barrier membrane. In group III, the defects were repaired using biodegradable barrier membrane seeded with cultured calvarial cell. Every surgical procedure were performed under the general anesthesia by using with intravenous injection of Pentobarbital sodium(30mg/Kg). After anesthesia, 5 rats were sacrificed by decapitation to obtain the calvaria for bone cell culture. Calvarial cells were cultured with Dulbecco's Modified Essential Medium contained with 10% Fetal Bovine Serum under the conventional conditions. The number of cell inoculated on the membrane were $1{\times}10^6$ Cells/ml. The membrane were inserted on the artificial bone defect after 3 days of culture. A single 3-mm diameter full-thickness artificial calvarial defect was made in each animal by using with bone trephine drill. After the every surgical intervention of animal, all of the animals were sacrificed at 1, 2, 3 weeks after surgery by using of perfusion technique. For obtaining histological section, tissues were fixed in 2.5% Glutaraldehyde (0.1M cacodylate buffer, pH 7.2) and Karnovsky's fixative solution, and decalcified with 0.1M disodium ethylene diaminetetraacetate for 3 weeks. Tissue embeding was performed in paraffin and cut parallel to the surface of calvaria. Section in 7${\mu}m$ thickness of tissue was done and stained with Hematoxylin-Eosin. All the specimens were observed under the light microscopy. The following results were obtained. 1 . During the whole period of experiment, fibrous connective tissue was revealed at 1week after surgery which meant rapid soft tissue recovery. The healing rate of defected area into new bone formation of the test group was observed more rapid tendency than other two groups. 2 . The sequence of healing rate of bone defected area was as follows ； test group, positive control, negative control group. 3 . During the experiment, an osteoclastic cell around preexisted bone was not found. New bone formation was originated from the periphery of the remaing bone wall, and gradually extended into central portion of the bone defect. 4 . The biodegradable barrier membrane was observed favorable biocompatibility during this experimental period without any other noticeable foreign body reaction. And mineralization in the newly formed osteoid tissue revealed relatively more rapid than other group since early stage of the healing process. Conclusively, the cultured bone cell inoculated onto the biodegradable barrier membrane may have an important role of regeneration of artificial bone defects of alveolar bone. This study thus demonstrates a tissue-engineering the approach to the repair of bone defects, which may have clinical applications in clinical fields of the dentistry including periodontics.

• Journal of the Korean Society of Radiology
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• v.5 no.1
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• pp.11-18
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• 2011

The periosteum contains multipotent cells that can differentiate into osteoblasts and chondrocytes. Cultured periosteum-derived cells (PDCs) have an osteogenic capacity. The purpose of this study was to evaluate the interaction of PDCs with bone graft biomaterial. After cell isolation from the calvarial periosteum of Sprague-Dawley rats, cultured PDCs were placed in critical-sized calvarial defects with beta-tricalcium phosphate (${\beta}$-TCP). All rats were sacrificed 8 weeks after bone graft surgery, and the bone regenerative ability of bone grafting sides was evaluated by plain radiography, micro-computed tomography (CT), and histological examination. PDCs grafted with ${\beta}$-TCP displayed enhanced calcification in the defect site, density of regenerated bone and new bone formation within the defect and its boundaries. Furthermore, these PDCs more efficiently regenerated new bone as compared to grafted ${\beta}$-TCP only. The results suggest that cultured PDCs have the potential to promote osteogenesis in bone defects.

• The korean journal of orthodontics
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• v.24 no.2
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• pp.295-301
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• 1994

The movement of teeth during orthodontic treatment requires bone remodeling process in periodontal tissue. To find out the changes occuring in the cell itself, mechanical force was applied to the cultured periodontal ligament cells. Following results were obtained from measuring the changes in cyclic AMP and $PGE_2$, $^3H$-thymidine incorporation amount in time lapse after application of mechanical force. 1. When mechanical force was applied to cultured PDL cells, the amount of cAMP in cells were increased significantly after 15 min. of force application, but were decreased gradually as time lapsed. 2. When mechanical force was applied to cultured PDL cells, the amount of PGE2 were increased at 20,40,60 min. and was significantly increased at 20 min. 3. When mechanical force was applied to cultured PDL cells, the amount of $^3H$-thymidine incorporation was some increased, but was not statistically significant.

• Proceedings of the Korean Society of Toxicology Conference
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• 2003.05a
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• pp.66-67
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• 2003

It is well known that osteoblasts and osteoc1asts playa key role in bone metabolism. They involve in osteoformation or bone destruction which are ragulated by various factors such as thyroid hormone, parathyroid hormone, estrogen, growth factor and cytokine. Recently, it is demonstrated that oxidative stress is one of pathological factors in bone metabolism, but it is left unknown about mechanism between oxidative stress and bone metabolism.(omitted)

• Journal of Periodontal and Implant Science
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• v.41 no.6
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• pp.293-301
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• 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.