• Journal of Periodontal and Implant Science
• /
• v.23 no.1
• /
• pp.77-96
• /
• 1993

This in vitro study was undertaken to observe whether citric acid application aids the attachment and proliferation of human periodontal ligament cells to the root surfaces of periodontally diseased teeth. The roots were prepared so that the comparison could be made among the control healthy root surface, citric acid demineralized and non-demineralized root planted surfaces. Prior to the cell attachment experiment, each groups were prepared for scanning electron microscopic (SEM) examinations of root surface morphology, All specimens were fixed with phosphate buffered glutaraldehydes, postfixed with phosphate buffered osmium tetraoxide and stained with phosphate buffered tannic acid. dehydrated in ethanol, critical point dried, sputter coated with gold and examined under the SEM. In the cell attachement experiment, human cultured periodontal ligament cells at concentration to $4.5{\times}\;10^4\;cells/ml$ were seeded in each culture well which contained prepared roots and incubated for 30min 1, 2, 6, 12 and 24 hours at 37, 5% $CO_2$air incubator. Than the specimens were prepared for SEM examination using, the same methods as described above. In the cell proliferation experiment, $5{\times}\;10^4\;cells/ml$ cells were seeded incubated with the specimens for 6 hours. Then, all of the specimens were moved into fresh culture well and incubated for 24, 48, and 72 hours. The cell counting was done after trypsinization, under light microscope. The results were as follows. When viewed the surface morphology prior to the cell attachment, the non acid treated root planed surface displayed scaling striation and occasional bacteria and calculus. The citric acid treated specimens displayed little debris on the surface and funnel shaped orifices of dentinal tubules. There were no apparent differences in the morphology of cells attached to the control and experiment groups. However, in initial attachement, there was a slight more enhanced appearance in attachment in citric acid treated groups than other root surfaces. After 6 hours of incubation, most of the cells initiated the alteration of cell morphology from ovoid to spindle shapes. After 24 hours of incubation, most of the cells displayed proliferated appearance and connected with each other via numerous processes. In the cell proliferation experiments, there were statistically significant increased number of cells in citic acid treated groups than other groups.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.32 no.3
• /
• pp.395-402
• /
• 2005

In considering the healing process of injured periodontal tissue, healing rate would be influenced by the cellular activity of periodontal fibroblasts(PDLFs). In addition, the reattachment among PDLFs should be induced for healing process. The purpose of this study was to evaluate the effects of epidermal growth factor(EGF) on the proliferation and attachment of PDLFs and to verify the efficacy of EGF as a storage media or a pre-replantation conditioner of traumatically avulsed tooth. Human recombinant epidermal growth factor(hrEGF) and human periodontal fibroblasts from first premolar were prepared. At first, MTT assay was done to evaluate the toxic effect on human periodontal fibroblast and the maximum cellular growth of EGF. Cellular proliferation rate was then compared between control group and 10ng/ml EGF added group. Also, western blot was done to evaluate the expression of fibronectin in both groups. The results were as follows: 1. From MTT assay, EGF showed no toxic effect on PDL fibroblasts. The highest proliferation was shown at 10ng/ml EGF. 2. In 10ng/ml EGF added group, the degree of proliferation of PDLFs was significantly higher than that in control group. 3. Fibronectin expression of EGF added group was also significantly higher than that of control group. From this study we could conclude that EGF enhanced the regeneration rate of periodontal fibroblast, which could be used as a pretreatment agent or a storage media for traumatically avulsed teeth.

• The korean journal of orthodontics
• /
• v.30 no.4 s.81
• /
• pp.441-452
• /
• 2000

The present studies were performed to investigate the interaction of $17{\beta}$-estradiol and human growth hormone(hGH) on the proliferation of human periodontal ligament(WDL) cell. The independent effects of $17{\beta}$ estradiol and hGH on hPDL cell proliferation were investigated and the effects of hGH on hPDL cell proliferation after $17{\beta}$-estradiol pre-treatment were also investigated. Lastly, the change of hGH receptor expression in hPDL cell after $17{\beta}$-estradiol pre-treatment were investigated. The obtained results were as follows; 1. The treatment of $17{\beta}$-estradiol or hGH had no significant effects on hPDL cell proliferation. 2. After pre-treatment of $17{\beta}$-estradiol, hGH stimulated the proliferation of the hPDL cell, regardless of hHG concentration. 3. Although there was not hGH receptor in the hPDL cell, hGH receptors were expressed in hPDL cell after more than 6 hours pre-treatment of $17{\beta}$-estradiol. 4. The effect of hGH on hPDL cell proliferation was related to the hGH receptor expression. $17{\beta}$-estradiol pre-treaaent contributed to the hGH effects on the hPDL cell by stimulating hGHR expression.

• The korean journal of orthodontics
• /
• v.28 no.4 s.69
• /
• pp.627-640
• /
• 1998

[$TGF-{\beta}$] is a polypeptide with multiple physiological functions in regulation of cell-to-cell interaction and in growth and development. The active form of vitmain $D_3$, 1,25-dihydroxycholecalciferol $[1,25-(OH)_2D_3]$, is one of the most potent stimulators of osteoclastic acitivity. The purpose of this study was to evaluate the effect of Vitamin $D_3$ and/or $TGF-{\beta}$ on the periodontal ligament(PDL) cells. Human PDL cells were prepared from the first premolars extracted for the orthodontic treatment and were incubated in the environment of , $37^{\circ},\;5\%\;CO_2\;and\;95\%$ humidity. 10, 50 or 100ng/m1 of $1,25-(OH)_2D_3$ and 0.1, 1, 5 or 10ng/ml of $TGF-{\beta}$ were administered to the culture wells, separately or in combination. And the viability of PDL cells was evaluated by MTT assay The obtained results were as follows. 1. The viability of PDL cells in 10ng/ml of vitamin $D_3$ was not significantly differenent from that of the control group at 1, 2 and 3-day of cultivation, but it was significantly increased in 50ng/ml of Vitamin $D_3$ at 3-day and in 100ng/m1 of Vitamin $D_3$ at 2 and 3-day. 2. The viability of PDL cells in 0.1ng/ml of $TGF-{\beta}$ was not significantly differenent from that of the control group at 1, 2 and 3-day of cultivation, but it was significantly increased in 1 and 5ng/ml of $TGF-{\beta}$ at 3-day of cultivation, and in 10ng/ml of $TGF-{\beta}$ at 2 and 3-day of cultivation. 3. In case of admixture of 1ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$, the viability of PDL cells was significantly increased in the admixture of 100ng/ml of vitamin $D_3$ at 3-day of cultivation 4. In case of admixture of 5ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$, the viability of PDL cells began to be increased from 2-day of cultivation in the admixture of 10 50 and 100ng/ml of vitamin $D_3$, but it was not maintained at 3-day in the admixture of 10ng/m of vitamin $D_3$. 5. In case of admixture of 10ng/ml of $TGF-{\beta}$ and the various concentrations of vitamin $D_3$ the viability of PDL cells was significantly increased in the admixture of 50ng/ml of vitamin $D_3$ at 2 and 3-day of cultivation, and in the admixture of 100ng/ml at 1, 2 and 3-day.

• The korean journal of orthodontics
• /
• v.27 no.6 s.65
• /
• pp.955-961
• /
• 1997

Nicotine is one of the major components of cigarette smoking which causes various systemic and local diseases to human body. Mitogenic effects of nicotine to systemic disease are interesting factors in the results of cellular Proliferation especially to vascular and pulmonary tissue or cells. The study of local effects concerns with destruction of tissue and delayed healing rate after various surgical treatment. Platelet-Derived Growth factor(PDGF) and Insulin-like growth factor(IGF) are blown as major mitogens to human PDL cells. The purpose of this study was to investgate the mitogenic effects of nicotine to human PDL cells. We studied the expression of PDGF-$\alpha$ receptor, PDGF-$\beta$receptor, and IGF-l receptor mRNA from the nicotine treated human PDL cells by northern analysis. The experimental groups were divided into different serum($1\%,\;10\%$) and nicotine (100ng/m1,1000ng/m1) concentrations and each group was studied by time course. The results of this study showed upregulation of PDGF-${\alpha},\;{\beta}$ receptor and IGF-l receptor mRNA at 100ng/ml nicotine concentration and $10\%$ serum group to the time course. These results suggest that physiologically attainable nicotine concentrations may stimulate the mitogenic gene synthesis to human PDL cells in vitro.

• The korean journal of orthodontics
• /
• v.31 no.4 s.87
• /
• pp.447-458
• /
• 2001

The purpose of this study was to evaluate 1) in vivo, the expression of chondroitin 4-sulfate (CH-4S), a structural element of glycosaminoglycans(GAGs), in periodontal tissue during the experimental movement of rat incisors, by labelled streptavidine biotin immunohistochemical staining for CH-4S, 2) In vitro, the expression of CH-4S in cultured human periodontal ligament(PDL) cells supplemented with 10ng/ml of $TGF-{\beta}_1$, 20ng/ml of PDGF-BB, 1ng/ml $TNF-\alpha$, or $1{\mu}g/ml$ LPS by western blot analysis. The results of this study were as follows ; 1. The expression of CH-4S was stronger in pulp, PDL, osteoblasts, osteoclasts and osteocytes in experimental group than in control group, but was rare in dentin, and cementum of experimental groups, regardless of the duration of force application, which was not different from that of control group. 2. In experimental group, the expression of CH-4S in pulp began to increase at 1 day after force application and got to the highest degree at 7 days. After 14 days, the expression in CH-4S immunoreactivity was decreased, and became similar to that of control group at 28 days. 3. The expression of CH-4S in PDL was noted in adjacent to alveolar bone. PDL showed higher intensity of immunolabelling after 1 day of orthodontic tooth movement. And the expression was more stronger in the tension side than that of pressure side of PDL at 1 day, but more stronger in the pressure side than that of tension side of PDL at 4 days. After 7 days, a decrease in CH-4S expression was observed. 4. The expression of CH-4S in alveolar bone got to the highest degree at 4 days, and At 7 days, a decrease in CH-4S expression was observed. 5. PDGF-BB notably raised the expression of CH-4S in the PDL cells at 3 days of cultivation 6. The expression of CH-4S of PDL cells was decreased with the application of $TNF-\alpha$ at 1 day. 7. Admixture of $TGF-{\beta}_1$ and PDGF-BB got more expression of CH-4S in PDL as compared to only $TGF-{\beta}1$ or PDGF-BB. A similar decrease of the expression of CH-4S was observed in the case of application of LPS or $TNF-\alpha$.

• The Journal of Korean Academy of Prosthodontics
• /
• v.54 no.3
• /
• pp.203-209
• /
• 2016

Purpose: Stimulating the proliferation and migration of periodontal ligament cells (PDLCs) has become the main goal of periodontal regeneration. To accomplish this goal, regeneration procedures have been developed, but results have not been predictable. Recently, tissue engineering using enamel matrix derivatives (EMDs) and growth factors has been applied to periodontal regeneration; however, the mechanism of EMDs is largely unknown. The aim of this study was to investigate the effects of EMDs on the proliferation and release of growth factors from PDLCs. Materials and methods: Human PDLCs were removed from individually extracted 3rd molars of healthy young adults, and cultured in the media containing EMDs (Emdogain, Biora, Malmo, Sweden) at concentration of 0, 12.5, 25, 50, 100, and $200{\mu}g/mL$ each. Cell proliferation and ALP (alkaline phosphatase) activity were measured. The evaluation of growth factors released by PDLCs was also performed by one-way analysis of variance (ANOVA) and Bonferroni's multiple comparison test. Results: Significantly increased proliferation and ALP activity were observed in PDLCs treated with over $25{\mu}g/mL$ and $50{\mu}g/mL$ EMDs, respectively. Additionally, treatment of PDLCs with $50{\mu}g/mL$ resulted in significantly increased release of vascular endothelial growth factor (VEGF) and transforming growth factor $(TGF)-{\beta}$ after 24 h and 48 h, respectively. Conclusion: EMDs enhance the proliferation and ALP activity of PDLCs, and promote the release of growth factors, including VEGF and $TGF-{\beta}$, from PDLCs. Therefore EMDs could be one of the effective methods for periodontal regeneration.

• Journal of Periodontal and Implant Science
• /
• v.22 no.2
• /
• pp.211.1-211.1
• /
• 1992

• 대한치주과학회:학술대회논문집
• /
• 2006.11a
• /
• pp.114-114
• /
• 2006

• Journal of Periodontal and Implant Science
• /
• v.32 no.3
• /
• pp.457-473
• /
• 2002

The purposes of this study is to evaluate the combination effects of TGF-${\beta}_1$ and PDGF-BB on the periodontal ligament cells to use 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/100% FBS at the $37^{\circ}C$, 5% $CO_2$ incubator. Authors measured the DNA synthesis, total protein, collagen and noncollagenous protein synthesis according to the concentration of TGF-${\beta}_1$,(1,5ng/ml) and PDGF-BB (1,10 ng/ml) in combination. To explore further this delayed effect of TGF-${\beta}_1$, we preincubated human periodontal ligament cells with TGF-${\beta}_1$ for 4 or 24 hours before PDGF-BB stimulation. The results were as follows: The DNA synthetic activity was increased dose dependently by TGF-${\beta}_1$, PDGF-BB. The combination of TGF-${\beta}_1$ and PDGF-BB consistently enhanced the DNA synthetic activity to PDGF-BB alone. The ability of TGF-${\beta}_1$ to enhance DNA synthetic activity in PDGF-BB treated periodontal ligament cells was dose dependent. The maximum mitogenic effect was at the 5ng/ml of TGF-${\beta}_1$ and l0ng/ml of PDGF-BB. Preincubation of cell with TGF-${\beta}_1$ resulted in significantly greater response to PDGF-BB at all TGF-${\beta}_1$ concentration studied, and may be useful for clinical application in periodontal regenerative procedures. The total protein, collagen and noncollagen synthesis was increased dose pendently by TGF-${\beta}_1$, PDGF-BB. The % of collagen was slightly decreased according to the concentration of TGF-${\beta}_1$, PDGF-BB. The effect of TGF-${\beta}_1$, PDGF-BB were not specific for collagen synthesis since it also increased noncollagenous protein synthesis. This study demonstrates that PDGF-BB is major mitogens for human periodontal ligament cells in vitro, and supports a role for TGF-${\beta}_1$ as a regulation of the mitogenic and total protein formation to PDGF-BB in these cells.