• The Journal of Advanced Prosthodontics
• /
• v.6 no.5
• /
• pp.406-414
• /
• 2014

PURPOSE. The objective of this study was to investigate the biologic effects of enamel matrix derivative (EMD) with different concentrations on cell viability and the genetic expression of human gingival fibroblasts (HGF) to zirconia surfaces. MATERIALS AND METHODS. Immortalized human gingival fibroblasts (HGF) were cultured (1) without EMD, (2) with EMD $25{\mu}g/mL$, and (3) with EMD $100{\mu}g/mL$ on zirconia discs. MTT assay was performed to evaluate the cell proliferation activity and SEM was carried out to examine the cellular morphology and attachment. The mRNA expression of collagen type I, osteopontin, fibronectin, and TGF-${\beta}1$ was evaluated with the real-time polymerase chain reaction (RT-PCR). RESULTS. From MTT assay, HGF showed more proliferation in EMD $25{\mu}g/mL$ group than control and EMD $100{\mu}g/mL$ group (P<.05). HGFs showed more flattened cellular morphology on the experimental groups than on the control group after 4h culture and more cellular attachments were observed on EMD $25{\mu}g/mL$ group and EMD $100{\mu}g/mL$ group after 24h culture. After 48h of culture, cellular attachment was similar in all groups. The mRNA expression of type I collagen increased in a concentration dependent manner. The genetic expression of osteopontin, fibronectin, and TGF-${\beta}1$ was increased at EMD $100{\mu}g/mL$. However, the mRNA expression of proteins associated with cellular attachment was decreased at EMD $25{\mu}g/mL$. CONCLUSION. Through this short term culture of HGF on zirconium discs, we conclude that EMD affects the proliferation, attachment, and cell morphology of HGF cells. Also, EMD stimulates production of extracellular matrix collagen, osteopontin, and TGF-${\beta}1$ in high concentration levels. CLINICAL RELEVANCE. With the use of EMD, protective barrier between attached gingiva and transmucosal zirconia abutment may be enhanced leading to final esthetic results with implants.

• Journal of Periodontal and Implant Science
• /
• v.31 no.3
• /
• pp.597-610
• /
• 2001

Normal gingival fibroblasts functioning is fundamental for the maintenance of periodontal connective tissue as well as wound healing. Nicotine have been found to affect DNA synthesis and cell proliferation, which appear to depend on the type of cells. This in vitro study was done to determine the effects of nicotine, a major component of tobacco, on cell proliferation, viability, activity, cell cycle distribution, and expression of cell cycle regulatory proteins in human gingival fibroblasts. Nicotine has been tested for 2 days or 4 days in 5 different concentrations; $0.1{\mu}g/ml$; $1{\mu}g/ml$; $10{\mu}g/ml$; $100{\mu}g/ml$; $1000{\mu}g/ml$. To assess cell proliferation and viability, viable and non-viable cells were counted by hemocytometer; to evaluate cellular activity, MTT assay was employed; to analyze cell cycle distribution, fluorescent propidium iodide-DNA complex were measured using fluorocytometer; to determine the expression of cell cycle regulatory proteins, western blot analysis was performed. After 2 days and 4 days incubation respectively, at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$, nicotine significantly inhibited proliferation comparing to non-supplemented controls. The cell viability was significantly decreased after 2 days and 4 days at concentrations of $1{\mu}g/ml$ - $1000{\mu}g/ml$ and at $10{\mu}g/ml$ - $1000{\mu}g/ml$ respectively. After 2 days and 4 days, the cellular activity was significantly decreased at concentrations of $10{\mu}g/ml$ - $1000{\mu}g/ml$. Treatment with $100{\mu}g/ml$ nicotine for 48 hours caused an increase in the proportion of G1-phase cells (from 46.41% to 53.46%) and a decrease in the proportion of S-phase cells (from 17.80% to 14.27%). The levels of cyclin $D_1$ and CDK 4 proteins in nicotine-treated fibroblasts were lower than that of controls, whereas the levels of p16 and pRB were higher than that of controls. These results suggest that the decrease of cell proliferation and lengthened Gap phases (G1) by nicotine may due to the increased expression of p16 and pRB as well as decreased expression of cyclin $D_1$ and CDK 4 in human gingival fibroblasts.

• Restorative Dentistry and Endodontics
• /
• v.31 no.4
• /
• pp.282-289
• /
• 2006

The purpose of this study was to compare the ability of three resin surface sealants to prevent microleakage in Class V composite resin restorations. Forty Class V cavities with the occlusal margin in enamel and gingival margin in dentin were prepared on the buccal surfaces of sound extracted molars, and restored with composite resin. Restorations were randomly assigned into one of four equal groups (n = 10): a control group, without resin sealing, and three experimental groups in which margins were sealed with Fortify Plus, Biscover and Permaseal, respectively. Specimens were thermocycled, immersed in a 2% methylene blue solution for 4 hours, sectioned longitudinally, and observed the leakage at the occlusal and gingival margins. The result was analyzed using Kruskal-Wallis test, Mann-Whitney test and Wilcoxon signed rank test. In conclusion, the ability to reduce microleakage at occlusal margins was similar in all of three sealants. However at gingival margin, it depended on the type of used resin surface sealant. At gingival margin. control and Fortify Plus group showed statistically higher microleakage than PermaSeal group. and Fortify Plus group also showed higher microleakage than BisCover group (p < 0.05).

• Journal of Periodontal and Implant Science
• /
• v.43 no.2
• /
• pp.87-95
• /
• 2013

Purpose: The purpose of this study was to analyze the expression of inducible nitric oxide synthases (iNOS), tissue inhibitors of metalloproteinase $(TIMP)_{-3}$, and $TIMP_{-4}$ in the gingival tissues of periodontal patients with or without type 2 diabetes mellitus (DM). Methods: Depending on the patient's systemic condition and clinical criteria of the gingiva, each gingival sample was classified into one of three groups. Sixteen clinically, systemically healthy patients (group 1), 16 periodontal patients (group 2), and 16 periodontal patients with DM (group 3) were included. Tissue samples in each group were collected, prepared, and analyzed by western blotting. Quantification of the relative amount of $TIMP_{-3}$, $TIMP_{-4}$, and iNOS was performed. Results: The expression levels of iNOS and $TIMP_{-3}$ both increased in group 1, group 2, and group 3 in increasing order, and were significantly higher in both group 2 and group 3 as compared to group 1 (P<0.05). The expression levels of $TIMP_{-4}$ increased in the same order, but significantly increased in group 2 as compared to group 1, in group 3 as compared to group 1, and group 3 as compared to group 2 (P<0.05). Conclusions: This study demonstrated that iNOS, $TIMP_{-3}$, and $TIMP_{-4}$ might be involved in the progression of periodontal inflammation associated with type 2 DM. It is thought that further study of these factors can be applied practically for the diagnosis and control of periodontitis in diabetics.

• Restorative Dentistry and Endodontics
• /
• v.16 no.1
• /
• pp.121-132
• /
• 1991

The purpose of the study was to evaluate the marginal sealing effect of composite resin inlays according to the luting techniques and compare them to the conventional direct resin filling technique. 90 cavities of class V were prepared on the buccal surface of 90 extracted molar teeth, which were divided into four groups. Cavities of control group were directly filled with Scotchbond 2 and P - 50, and those of composite resin inlay groups were luted with one of the followings: Adhesive bond followed by Adhesive cement, All bond followed by Adhesive cement, Fuji - ionomer type L All the specimens were immersed in India ink dye solution for 7 days at $37^{\circ}C$ incubator after thermocycling between $5^{\circ}C$ and $60^{\circ}C$ and longitudinally sectioned with diamond disk inot two parts All the specimens were observed at the occlusal and gingival margins and statistical analysis was performed. The results were as follows: 1. Groups filled with composite resin inlay showed less marginal leakage than the group directly filled(p<0.01). 2. There was no significant difference in marginal leakage between composite resin inlay groups luted with Adhesive bond followed by Adhesive cement and the group luted with All bond followed by Adhesive cement(p>0.05). 3. At occlusal margins, Composite resin inlay group luted with Adhesive bond followed by Adhesive cement showed less marginal leakage than the group luted with Fuii ionomer type I(p<0.01). At gingival margins, composite resin inlay group luted with All bond followed by Adhesive cement showed less marignal leakage than the group luted with Fuji ionomer type I(P<0.01).

• Journal of Periodontal and Implant Science
• /
• v.44 no.4
• /
• pp.158-168
• /
• 2014

Purpose: The present study was conducted to evaluate the impact of scaling and root planing (SRP) on the C-reactive protein (CRP) levels of gingival crevicular fluid (GCF) and serum in chronic periodontitis patients with type 2 diabetes mellitus (T2DM-CP) or without type 2 diabetes mellitus (NDM-CP). Methods: Forty-eight human participants were divided into two groups: an experimental (T2DM-CP) group (group I, n=24) comprising chronic periodontitis patients with random blood sugar ${\geq}200mg/dL$ and type 2 diabetes mellitus, and control (NDM-CP) group (group II, n=24) of those with chronic periodontitis and random blood sugar <200 without T2DM for the study. All subjects underwent nonsurgical periodontal therapy (NSPT) including complete SRP and subgingival debridement. Periodontal health parameters, plaque index (PI), gingival index (GI), probing pocket depth (PPD), clinical attachment level (CAL), GCF volume (GCF vol), GCF-CRP, random blood glucose (RBS), glycated hemoglobin, and systemic inflammatory markers, serum CRP, total leukocyte count (TLC), neutrophil count (Neutr) and lymphocyte count (Lymph), were evaluated at baseline, 1 month, and 3 months after SRP. Results: NSPT resulted in statistically significant improvement in periodontal health parameters (PI, GI, PPD, CAL, GCF vol), CRP levels in serum as well as GCF of both groups I and II. The mean improvement in periodontal health parameters (PI, GI, PPD, CAL, GCF vol), CRP levels in serum and GCF was greater in group I than group II after NSPT. There was nonsignificant increase in GCF-CRP, TLC, Lymph, and RBS, and a significant increase in Neutr and Serum CRP in group II at 1 month. The Serum CRP level of 20 out of 24 group II patients had also increased at 1 month. Conclusions: The CRP levels in both GCF and serum were higher in T2DM-CP patients than in NDM-CP patients. Although there was a significant improvement in both the groups, greater improvement was observed in both GCF and serum samples of T2DM-CP patients.

• Journal of Periodontal and Implant Science
• /
• v.36 no.3
• /
• pp.613-625
• /
• 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 Periodontal and Implant Science
• /
• v.37 no.4
• /
• pp.755-766
• /
• 2007

Purpose: The purposes of this study were to compare and quantify the expression of $PGE_2$, MMP-14 and TIMP-1 in the gingival tissues of patients with type 2 diabetes mellitus and healthy adults of chronic periodontitis with alveolar bone resorption. Material and methods: Gingival tissue samples were obtained during periodontal surgery or tooth extraction. According to the patient's systemic condition & clinical criteria of gingiva, each gingival sample was devided into three groups. Group 1 (n=8) is clinically healthy gingiva without bleeding and no evidence of bone resorption or periodontal pockets, obtained from systemically healthy 8 patients. Group 2 (n=8) is inflammed gingiva from patients of chronic periodontitis with alveolar bone resorption. Group 3(n=8) is inflammed gingiva from patients of chronic periodontitis with alveolar bone resorption associated with type 2 diabetes. Tissue samples were prepared and analyzed by Western blotting. The quantification of $PGE_2$ MMP-14 and TIMP-1 were performed using a densitometer and statistically analyzed by one-way ANOVA followed by Tukey test. Results: The expressions of MMP-14 and TIMP-1 were showed increasing tendency in group 2 & 3 compared to group 1. The expressions of $PGE_2$, MMP-14 were showed increasing tendency in group 3 compared to group 1 and group 2. According to MMP-14 levels were increasing, $PGE_2$ showed increasing tendency in group 3, and although $PGE_2$, MMP-14 levels were increasing, TIMP-1 levels were similar expressed comparing to group 2. Conclusion: In conclusion, this study demonstrated that the expression levels of MMP-14 and TIMP-1 had increasing tendency in inflammed tissue. It can be assumed that $PGE_2$ and MMP-14 may be partly involved in alveolar bone resorptive process and the progression of periodontal inflammation associated to type 2 DM.

• The Journal of Korean Academy of Prosthodontics
• /
• v.18 no.1
• /
• pp.37-47
• /
• 1980

The purpose of this study was to investigate the stresses in different proximal margins and to measure, quantitatively, the effect of different modifications in the design of preparations on the stresses using two-dimensional photoelasticity. Photoelastic stress analysis is based on the phenomenon, exhibited by most transparent solids, of becoming birefringent, or doubly refracting, when strained. Two birefringent materials were used in this study, PSM-1 and PSM-5 in .standard sheet ($10'{\times}10'{\times}\frac{1}{4}'$ thickness), PSM-1(polyester) was used for constructing the substructure, and PSM-5(epoxy resin) was used in making the restorations to be investigated. Two birefringent materials were used in the construction of composite photoelastic model. Seven variable models were constructed. The peripheral dimensions of all model were constant and the models represent an occlusomesial section of a lower posterior molar. Model 1 represents the knife edge margin (shoulderless), Model 2 represents the chamfer, Model 3 represents a rounded shoulder(no sharp angle between the axial wall and gingival floor), Model 4 represents a flat shoulder (axial wall is a $90^{\circ}$ angle to the gingival wall), Model 5 represents $+15^{\circ}$ angulation, Model 6 has a $-15^{\circ}$ angulation, and Model 7 is the same as Model 4 except that it has a $45^{\circ}$ bevel. Improved artificial stone was used to represent dental cement in luting the composite photoelastic model. Static loading procedures(100 pounds) were used at preplanned sites. The results were as follows; 1. The stresses in the proximal portion of all tested models were compressive in nature when the proximal shoulders were loaded vertically on the same proximal marginal ridge. 2. The round and chamfered preparations were the optimum designs in proximoocclusal restorations. They showed the lowest stress concentration factor, i.e. 2.16 and 2.23, respectively. The knife edged shoulder had the highest value, K=5.39. Round type shoulder geometry experiments reduced the stress concentration factor (S.C.F.) 3. The gingival portion of proximal shoulder geometry was a critical location for stress concentration.

• The Journal of Internal Korean Medicine
• /
• v.16 no.2
• /
• pp.146-177
• /
• 1995

The purpose of this study was to investigate the relationship between the teeth and Zhang-Fu(臟腑), dental diseases, and the hygiene of the mouth through the literature of oriental medicine. First, the relatonship between the teeth and Zhang-Fu is reviewed as follows: The teeth are influenced by Shen(腎) because they are the end of bone and Biao(標) of Shen. Gingiva is related to Wei(胃) and Da-Chang(大腸) because it is passed by Yangming-Channel(陽明經). The growth and nutrition of teeth depends on Shen. The pathological condition of Shen causes the gingival atrophy, the loose of teeth, dedentition due to aging, withering of teeth, and tartar: whereas the pathological condition of Wei and Da-Chang causes toothache, gingivitis, inflamed gums, bad breath, and gingival hemorrhage. Second, the causes and therapies of dental diseases through the literature can be summarized as follows: The major causes of toothache are the pathogenic condition of wind-heat and wind-cold, the heat syndrome of Wei, the damp-heat of intestine, flaring-up of fire of deficiency type, rotten tooth, etc... The principal causes of dedentition and the shaking and loose of teeth are the deficiency of Shen, and the rest of causes are the damp-heat of Yangming. Gingival atrophy is caused by the deficiency of Shen, whereas the gingival hemorrhage comes from the factors in the pathogenic factor of wind-heat of Yangming-Channel, the heat syndrome of stomach, and the deficiency of Shen. The causes of grinding of teeth during sleeping are stomach-heat, and the delayed dentition and the withering result from the deficiency of Shen-Jing.(腎精) The principal therapies of toothache are removing wind and heat, clearing away heat and prompting diuresis, clearing away the stomach-heat, replenishing vital essence to tonify the Shen, relieving superficial syndrome by wind-cold, and alleviating pain by destroying parasites. For the prescription of the principal therapies, there are Xijio Dihuang Tang, Jiajian Ganlu Yin, Qufeng Wan, Qingwei San, Tiaowei Chenggi Tang Shengong Wan, Liangge San Qingwei Tang Yunu Jian, Liuwei Dihuang Wan Zuogui Yin Bawei Wan Wanshao Dan, Xixin San Badou Wan Gianghuo Fuzi Tang, Jiuzi Tang Badou Wan, etc... The therapies of dedentition and the shaking and loose of teeth are replenishing vital essence to tonify the Shen, and warming and recuperating the Shen-Yang: as the prescription, there are Liuwei Dihuang Wana Zuogui Yin, and Bawei Wan Anshen Wan Wanshao Dan Yougui Wan etc... The therapies of gingival hemorrhage are clearing away the stomach-heat, replenishing vital essence to tonify the Shen, warming and recuperating the Shen-Yang(腎陽), and moisturing and purging intence heat with the prescription of Tiaowei Chenggi Tang Xijiao Dihuang Tang, Liuwei Dihuang Wan Zuogui Yin, Bawei Wan Anshen Wan, and Yunu Jian. The therapy of gingival atrophy is replenishing vital essence to tonify the Shen in the prescription of Liuwei Wan Bawei Wan Ziyin Dabu Wan. The therapies of grinding of teeth during sleeping are clearing away the stomach-heat and purging intense heat, and invigorating the spleen through eliminating dampness in the prescription of Qingwei San, Wumei Wan, etc... The therapy of delaed dentition is replenishing vital essence to tonify the Shen with the prescription of Liuwei Wan Buyin Jian, etc... Third, clinical treatment reports of dental diseases are reviewed as follows: The toothache due to stomach-heat was treated by medical herbs like Gypsum, Natrir Sulfas, Rehmanniae, Schizonepetal Herba, Menthae Folium, Cimicifugae Rhizoma, and Scrophulariae Radix. The therapies of toothache due to flaring-up of fire in deficiency type from deficiency of Shen provided with replenishment of vital essence to tonify the Shen and clean ministerial fire, and the prescription was the kind of Liuwei Wan, which worked very well. The therapy of dedentition and loose of teeth due to deficiency of Shen was done to stablize the teeth as tonifing the Shen with the prescription of Guchi Wan. The rate of imrovement was over 90%. The destruction of periodontal tissue due to periodonititis was cured of dispelling wind, reducing heat, and alleviating pain, It was improved by taking Zizhi Xingiong Tang, Guchi Xiaotong San, Yunii Jian, and Qingwei San about 3-7 days, and the rate of improvement was over 80%. Fourth, the prevention and regimens are reviewed as follows: As a physical and breathing exercise of the teeth, tapping teeth which stimulates the circulation of Qi(氣) and Xue(血) had been used. The tapping time of 14, 17, 36, etc... has been reported, and it should be applied based on the body condition. The medical herbs for gargling and brushing teeth have been used. Specifically, Cimicifugae Rhizoma, Gypsum, Gypsum Fibrosum, and Indigo pulrelrata Lereis have been used to reduce heat, Coptidis Rhizama and Yang Jinggu to eliminate damp-heat, Amomi Semen, Cyperi Rhizoma, Flos Caryophylli, Asari Radix, Piperis Longi Fructus, Santali Albae Lignum, Meliae Fructus, Moschus, Aquillaiae Lignum, and Borneol to promote the circulation of Qi and to relieve pain, Ligustici Radix, Angelice Radix, Rhizoma Nardostachydis, Tribuli Semen to relieve superficial syndrome by means of diaphiresis, and Cnidii Rhizoma, Angelicae sinensis Radix, and Olibanum to promote blood circulation to stop pain.