• Journal of the korean academy of Pediatric Dentistry
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• v.34 no.1
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• pp.36-42
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• 2007

To compare the survival rate of periodontal ligament cells preserved in storage media with good availability at the time of an avulsion injury, periodontal ligament cells were incubated in ${\alpha}-MEM$ culture medium containing 10% FBS in condition of $37^{\circ}C$, 5% $CO_2$. These cells were then cultured in HBSS, ${\alpha}-MEM$, milk(S co., P. co.) and tap water at the temperature of 4, 25, $37^{\circ}C$ each in 60 min. The groups were measured by MTT assay. The results were as follows : 1. Among the storage media at $4^{\circ}C$, ${\alpha}-MEM$ and P-milk had the highest preserving ability of periodontal ligament cells, while that of HBSS S-milk and tap was low in order. 2. Among the storage media at $25^{\circ}C$, ${\alpha}-MEM$ had the highest preserving ability of periodontal ligament cells, while that of P-milk, HBSS, S-milk, tap water was low in order. 3. Among the storage media at $37^{\circ}C$, the preserving ability of periodontal ligament cells was very high in ${\alpha}-MEM$, P-milk, HBSS and S-milk, it's lowest in tap water. 4. The preserving ability of periodontal ligament cells in ${\alpha}-MEM$ was high at $4^{\circ}C$ and it's low in order of $25^{\circ}C$, $37^{\circ}C$, but in HBSS was high at $4^{\circ}C$ and it's low at $25^{\circ}C$, $37^{\circ}C$ 5. The preserving ability of periodontal ligament cells in S-milk and P-milk was high at $4^{\circ}C$, $25^{\circ}C$ and it s low at $37^{\circ}C$. In conclusion, HBSS is the storage medium of choice in an avulsion, but in this study it is preferable to choose milk at $4^{\circ}C$ for tooth since it is easy to get and affect cell viability.

• Journal of Periodontal and Implant Science
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• v.36 no.3
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• pp.613-625
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• 2006

Periodontal ligament(PDL) cells and human gingival fibroblasts(HGFs) play important roles in development, regeneration, normal function, and pathologic alteration. PDL cells and HGFs have the similarity related with general characteristics of fibroblast such as spindle shaped morphology, the presence of vimentin intermediate filament and the synthesis of interstitial collagens and fibronectin. There were many studies about the differences between PDL cells and HGFs, but they were not about whole gene level. In this study, we tried to explain the differences of gene expression profiles between PDL cells and HGFs, and the differences among three individuals by screening gene expression patterns of PDL cells and HGFs, using cDNA microarray. Although there were some variants among three experiments, a set of genes were consistentely and differentially expressed in one cell type. Among 3,063 genes, 49 genes were more highly expressed in PDL cells and 12 genes were more highly expressed in HGFs. The genes related with cell structure and motility were expressed more highly in PDL cells. These are cofilin 1, proteoglycan 1 secretory granule, collagen type I(${\alpha}$ 1), adducin gamma subunit, collagen type III(${\alpha}$ 1), fibronectin, lumican(keratan sulfate proteoglycan), and ${\alpha}$ -smooth muscle actin. Tissue inhibitor of metalloproteinase known as the enzyme controlling extracellular matrix with matrix metalloproteinase is more highly expressed in PDL cells, osteoprotegerin known as osteoclastogenesis inhibitory factor is more highly expressed in HGFs. We performed northern blot to verify cDNA microarray results on selected genes such as tissue inhibitor of metalloproteinase, fibronectin, osteoprogeterin. The result of northern blot analysis showed that each cell expressed the genes in similar pattern with cDNA microarray result. This result indicates that cDNA microarray is a reliable method in screening of gene expression profiles.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.1
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• pp.157-164
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• 2003

Cervi Parvum Cornu(CPC) is that the young horn of deer family and has been traditionally used as a medicine in Eastern. The purpose of present study was to investigate the effects of CPC on cell cycle progression and its molecular mechanism in human periodontal ligament cells (HPOLC). In cell proliferation assay, 1 ng/ml, 10 ng/ml, 100 ng/ml, 1 ㎍/ml and 10 ㎍/ml of CPC were used, all treatment groups increased the cell growth. Maximal cell proliferation was observed in cells exposed to 100 ng/ml of CPC at 4 day, and 10 ng/ml and 100 ng/ml of CPC at 6 days. S phase was increased and G1 phase was decreased in the group treated with 100 ng/ml of CPC in cell cycle analysis. The protein levels of cyclin D1 were not changed, but the levels of cyclin E, cdk 2, cdk 4 and cdk 6 were increased. The protein levels of p21, pRb were decreased as compared to that of control group, but the levels of p53 was not changed in the cells both treated with CPC Md untreated. These results suggested that CPC increases the cell proliferation and cell cycle progression in HPDLC, which is linked to an increased cellular levels of cyclin E, cdk 2, cdk 4 and cdk 6, and decreased the levels of p53, p21.

• The korean journal of orthodontics
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• v.24 no.4 s.47
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• pp.871-883
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• 1994

This experiment was designed to study possible roles of $interleukin-1\beta$, interleukin-6 and tumor necrosis $factor-\alpha$ in bone remodeling by measuring their effects on $PGE_2,\; LTB_4$ and collagenase production when they were administered to human periodontal ligament fibroblasts. Human periodontal ligament fibroblasts were collected from first premolars extracted for orthodontic treatment. They were incubated in the environment of $37^{\circ}C,\;5\%\;Co^2,\;and\;100\%$ humidity. They were treated with $0.25\%$ trypsin-EDTA solution and centrifuged. PDL cells in the fifth to seventh passage were used for the experiment. Cells were seeded onto the culture dishes and when they were successfully attached, human recombinant $interleukin-1\beta$, interleukin-6, and tumor necrosis $factor-\alpha$ were administered, alone or in combination. They were incubated for 4, 8 and 24 hours and the levels of $PGE_2,\;LTB_4$ and collagenase released into the culture media were assessed by enzymeimmunoassay and collagenase activity assay. The conclusions are as follows: 1. $IL-1\beta\;and\;TNF-\alpha$ were very active in stimulating the production of $PGE_2$ and collagenase by human periodontal ligament fibroblasts, while IL-6 increased $LTB_4$ production. 2. $IL-1\beta$ significantly increased $PGE_2$, but $LTB_4$ Production was not increased. $IL-1\beta$ is thought to act mainly via the cyclooxygenase pathway of arachidonic acid metabolism. 3. IL-6 tended to inhibit $IL-1\beta$ in the production of $PGE_2$ and collagense whereas IL-6 and $TNF-\alpha$ showed auditive effect in the level of $PGE_2$. The above cytokines increased the release of at least one of $PGE_2,\;LTB_4$ and collagenase. It suggests that cytokines are involved in bone remodeling process by stimulating PDL fibroblasts to produce various bone-resorptive agents. The roles of cytokines in bone remodeling as a whole would need further study.

• Journal of the korean academy of Pediatric Dentistry
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• v.26 no.2
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• pp.427-436
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• 1999

Preservation of the remaining periodontal ligament cells on an avulsed tooth is very important to the successful outcome of replantation. HBSS is recommended as the most suitable storage medium for the avulsed tooth that cannot be replanted immediately. But their availability near the site of an accident is doubtful. The purpose of this in vitro study was to compare periodontal ligament cells stored in different storage media obtained easily on the spot. Human periodontal ligament cells were collected from the premolar teeth extracted for orthodontic treatment. Cells were cultured in ${\alpha}-MEM$ culture medium containing 20% FBS, at $37^{\circ}C$ 100% humidity, in a 5% $CO_2$ incubator. Cells were cultured in 96 well culture plate, $5{\times}10^4$ cells per well with ${\alpha}-MEM$ and incubated for 24 hours. After discarding the medium, those cells were cultured in ${\alpha}-MEM$ contained with 10% FBS, pasteurized milk, sterilized saline, unstimulated saliva and bench-dried state at $25^{\circ}C$ room temperature for 30, 60, 90, 120, 180 minutes respectively. And then each group was measured using MTT assay. The results were as follows. 1. Between the group of each time, there was statistically significant difference. Periodontal ligament cells viability was highest in pasteurized milk and was reduced stepwisely in sterilized saline, unstimulated saliva and bench-dried state(p<0.05). 2. between the time of each group, there was statistically significant difference(p<0.05) but was no statistically significant difference at 90-120 minutes in pasteurized milk and at 60-90 minutes and 120-180 minutes in sterilized saline(p>0.05). In conclusion, HBSS as storage medium of an avulsed tooth is not practical on the spot. Insteadily pasteurized milk can be recommended to maintain the periodontal ligament cells viability.

• The korean journal of orthodontics
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• v.22 no.1
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• pp.273-283
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• 1992

To find out the differences between periodontal ligament cells (PDL cells) and gingival fibroblast cells (GFB cells), alkaline phosphatase, a marker enzyme for osteoblast, was used to measure the activities and $^{45}CaCl_2$ isotope was used to find out cellular and release of $^{45}Ca$, a requisite for bone formation,. PDL cells and GFB cells from 1 to 5 passages were also measured in alkaline phosphatase activity assay. By the use of above methods, followings were concluded that the PDL cells and the GFB cells have characteristics that are different from each other. In that PDL cells showed large amount of calcium uptake and large amount of calcium release in initial stage, they seem to possess characteristics which are similar to osteoblast-like cells. 1. The PDL cells, in contrast to the gingival fibroblast, showed exceedingly high alkaline phosphatase activity which was highest at the second passage, decreasing thereon. But gingival fibroblasts cells showed no distinct differences in alkaline phosphatase activity as the passage were elapsed. 2. For both PDL cells and GF cells, the $^{45}Ca$ uptake was greatest at 2 hours period. The PDL cells showed higher measuring than GFB cells through out the whole time period. 3. Whereas the GFB cells showed slow increase of $^{45}Ca$ release as time relapsed, the PDL cells showed rapid increase of $^{45}Ca$ release.

• The korean journal of orthodontics
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• v.28 no.1 s.66
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• pp.143-153
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• 1998

Platelet-derived growth factor(PDGF) and lipopolysaccharide(LPS) may be the important regualtors of bone metabolism Exogenous PDGF is recognized to have a stimulating effect on bone resorption in organ culture but to stimulate the formation of new bone ultimately. LPS is known to be a stimulating agent on the osteoclastic activity. The purpose of this study was to evaluate the effects and the interaction of PDGF and LPS on periodontal ligament(PDL) cells which have important roles in bone remodeling. Cultured human periodontal ligament cells were tented with various concentration or PDGF and/or LPS. The cellular viability was measured by Microtitration(MTT) assay according to the lapse time of culture. The obtained results were as follows: 1. The viability of PDL cells was not different from the con01 in 0.1ng/ml of PDGF, but was significantly increased to be over the level of control in 1ng/ml of PDGF at the second day of culture, and in 10ng/m1 of PDGF at the second and the third day of culture. 2. The cellular viability was decreased in $0.5{\mu}g/ml$ or $5{\mu}g/ml$ LPS at the third day of culture. 3. Incubation with both 1ng/ml or 10ng/ml of PDGF and $0.5{\mu}g/ml$ of $5{\mu}g/ml$ of LPS resulted in the increased cellular viability at the third day, which was greater than LPS only treated group. It was greater than even the control group in 10ng/m1 of PDGF. From the above findings, we could summarize that the admixture of PDGF and LPS could not less increase the viability of the human periodontal ligament cells than PDGF only.

• Journal of Periodontal and Implant Science
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• v.22 no.2
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• pp.212-212
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• 1992

• Journal of Periodontal and Implant Science
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• v.26 no.2
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• pp.396-413
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• 1996

Current acceptable methods for promoting periodontal regeneration are based on removal of diseased soft tissue. root treatment, guided tissue regeneration, introduction of new graft materials and biological mediators. Insulin-like growth factor-I(IGF-I) and Platelet-derived growth factor-BB(PDGF-BB), the members of the polypeptuyde growth factor family have been reported as the biological mediators which regulate a variety cellular matrix biologic activities of wound healing process including the cell proliferation, migration and extracellular matrix synthesis.The purposes of this study is to evaluate the combination effects of IGF-I and PDGF-BB on the cellular activity of the periodontal ligament cells to act as a regeneration promoting agent of periodontal tissue. Human periodontal ligament cells were prepared from the first premolar tooth extracted for the orthodontic treatment and were cultured in DMEM containing 10% FBS at the $37^{\circ}C$, 5% CO2 incubator. Author measured the DNA synthetic activity, and total protein, collagen and noncollagenous protein synthetic activities according to the concentration of 10,100ng/ml IGF-I and1,10 ng/ml PDGF-BB in combination. The results were as follows: Significantly increased in the 1 ng/ml PDGF-BB alone compared to the 10 ng/ml PDGF-BB alone(P<0.01) and in the 1 ng/ml PDGF-BB and 10, 100ng/ml IGF-I in combination compared to the 1 ng/ml PDGF-BB alone(P<0.05, P<0.0l). The synthetic activity of the total protein and collagen is significantly increased like to the synthetic activity of the DNA(P<0.05). The synthetic activity of the noncollagenous protein is increased according to the concentration of IGF_I, but not statistically statistically significant(P>0.05). The percent of the collagen is significantly in the 1ng/ml PDGF-BB and 10ng/ml IGF-I in combination compared to the 1ng/ml PDGF-BB alone(P<0.05) and in the 10ng/ml IGF-I in combination compared to the 10ng/ml PDGF-BB alone(P<0.05). The synthetic activity of the DNA is In conclusions, the percent study shows that PDGF-BB and IGF-I in combination have a potentiality to enhance the DNA synthesis and the total protein and collagen synthesis of The periodontal ligament cells, especially it is more significant in the low concentration of PDGF-BB compared to the high one. Thus, the PDGF-BB and IGF-I in combination may have important roles in promotion of periodontal litgment healing, and consequently, may useful for clinical application in periodontal regenerative procedures.

• Journal of Periodontal and Implant Science
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• v.28 no.2
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• pp.235-247
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• 1998

Calcium sulfate has a long history of medical use as an implant material. The biocompatibiliry of the material has been clearly established. Bone ingrowth concomitant with resorption occurs rapidly with efficient conduction of bone from particle to particle. Calcium sulfate also has a potential for functioning as a good bamer membrane. The purpose of this study was to compare the biocompatibility of different types of calcium sulfate grafting materials including an expelimental calcium sulfate compound on periodontal ligament cells in vitro as a preliminary test towards the development of a more convenient and useful form of grafting material which could promote regeneration of periodontal tissue. Human periodontal ligament cells were collected from the premolar teeth extracted for orthodontic treatment. cells were cultured in a.MEM culture medium containing 20% FBS, at $37^{\circ}C$ and 100% humidity, in a 5% CO2 incubator. Cells were cultured into 96 well culture plate $1{\times}104$ cells per well with $\alpha$-MEM and incubated for 24 hours. After discarding the medium, those cells were cultured in $\alpha$-MEM contained with 10% FBS alone (control group), in medcal-grade calcium sulfate(MGCS group), in plaster(plaster group), experimental calcium sulfate paste(CS paste group) for 1, 2, 3 day respectively. And then each group was characterized by examining of the cell counting, MTI assay, collagen synthesis. The results \vere as follows. 1. In the analysis of cell proliferation by cell counting, both medical-grdde calcium sulfate group and plaster group showed no stastically significant difference at day 1, 2, 3 accept for plaster group at day 1 compared to control group, but there was stastically significant difference between CS paste group and all other groups at day 1, 2, 3(P<0.05). 2. In the analysis of cytotoxicity by MIT assay, both medical-grade calcium sJlfate group and plaster group showed no stastically significant difference compared to control group at day 1, 2, 3 but there was stastically significant difference between CS paste group and all other groups at day 1, 2, 3(P<0.OS). 3. In the analysis of collagen synthesis by immunoblotting assay, high level was detected for medical-grade calcium sulfate group and plaster group at day 1, 2, 3 compared to CS paste group. On the basis of these results, medical-grade calcium sulfate and plaster was shown to possess biocompatibility whereas the CS paste had unfavourable outcome. This observation shows a need for modification of the materials contained in calcium sulfate paste.