• The Journal of Korean Academy of Prosthodontics
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• v.41 no.2
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• pp.111-127
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• 2003

Statement of problem : Resin cements were used widely on all ceramic crowns, but the influence of resin cements on biocells was not understood clearly. Purpose : This study was investigated to evaluate the biocompatibility of resin cements for all-ceramic crowns. Material and Method : The resin cements used in this study were Panavia F (Kuraray Co., Ltd. Japan), Variolink II (Vivadent Ets., Schann / Liechtenstein), and Bistite II (Bistite dual cure resin cement-clear Tokuyama Soda Co. Japan). The viability of normal human oral keratocytes, gingival fibroblast, and gingival fibroblast immortalized by Human Papilloma virus 16 was measured in vitro for evaluation of cytotoxicity on resin cements, and the response of pulp tissue was analyzed and evaluated with light microscope after application of cements at cutting edge of incisors. Results : The normal human oral keratocytes was the most sensitive to toxicity of resin cement, and toxicity of cements was higher in Bistite II than in Variolink II. The cell viability of immortalized gingival fibroblast did not affected by type of cement and cultivation period, but there was a tendency that cytotoxicity in Bistite II was higher than in Variolink II. The cell viability of gingival fibroblast was similar to that of immortalized gingival fibroblast regardless of cement type, but Bistite II showed more toxic than others after 5 days cultivation. The responses of pulp tissue according to cement type were similar after 2 days cultivation, but revealed high toxicity in Bistite II after 10 days cultivation. Conclusion : Variolink II was more biocompatible than any other resin cements used in this study.

• Biomedical Science Letters
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• v.10 no.3
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• pp.269-273
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• 2004

This study was designed to investigate the effect of ferulic acid on cell viability and cell adhesion activity in normal human gingival fibroblasts. The cell viability and cell adhesion activity of ferulic acid was measured by MTT assay or XTT assay, respectively, after normal human gingival fibroblasts were treated with or without ferulic acid for 48 hours. The cell viability of ferolic acid on normal human gingival fibroblasts did not show any decreasement by MTT assay and also, cell adhesion activity did not decreased by XTT assay, respectively, compared with control after cells were treated with various concentrations of ferolic acid for 48 hours. MTT/sub 50/ and XTT/sub 50/ were 2,130.0 μM and 1,773.7 μM ferolic acid, respectively. These results suggest that ferolic acid is non-toxic to normal human gingival fibroblasts by showing no significant differences in the cell viability and the adhesion activity compared with control by colorimetric assay.

• Journal of Periodontal and Implant Science
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• v.32 no.3
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• pp.603-614
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• 2002

Human gingival fibroblasts have proven to useful as a species specific cell culture system in various system on periodontal disease and regeneration. However, their use is limited, since they are hard to obtain and lifespan is short due to replicative senescence. To overcome these disadvantages, we transfected primary human gingival fibroblasts by the E6 and E7 genes of the Human papilloma virus(HPV) 16. The full length of HPV 16 E6 and E7 was cloned from the pBR322 into BamHl and Sal I of a pBabe vector including hygromycin B resistance. Before pBabeE6/E7 plasmid transfection, peak 8 GFP including G418 resistance was transfected into primary GF to check the transfection efficency. PBabe E6/E7 plasmid was transfected using Lipofectamine plus following manufacter's instruction into primary normal human gingival fibroblasts in 60mm dishes with FBS free DMEM. After 2 days of transfection, the cells were treated with hygromycin for 2 weeks until the transfected control cells died. The resulting hygromycin resistant colonies were pooled, and clonned, and sucessful　transfection was established for immortalized gingival fibroblast cell lines. Immoralized GF cells showed stellate shape, that is similar to that of orange grains, and more rapid growth and higher proliferation than that of primary gingival fibroblasts. This cell lines overcame crisis and could be cultured over 30 subcultured, could be use for three dimentional culture, epithelial-mesenchymal interaction study.

• International Journal of Oral Biology
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• v.32 no.2
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• pp.67-73
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• 2007

Periodontitis is a chronic infectious disease that leads to periodontal destruction, and is one of the major causes of tooth loss in humans. The osteoclast differentiation factor (ODF), which is also known as the receptor activator of the NF-kB ligand (RANKL), is a surface-associated ligand on bone marrow stromal cells and osteoblasts. RANKL activates its cognate receptor, RANK, on osteoclast progenitor cells, which leads to the differentiation of mononucleated precursor cells. Osteoprotegerin (OPG) is a decoy receptor that is released from stromal cells and osteoblasts to inhibit the interaction between RANKL and RANK. Although the precise mechanism of bone loss in periodontitis is unknown, the differentiation and activation of osteoclasts by OPG-ODF-RANK signaling might play the role in periodontal bone destruction. The relationship between the concentration of sex hormones and the expression of ODF and OPG was examined by treating human gingival fibroblasts and periodontal ligament cells with the normal serum concentration of estrogen or progesterone during menstruation or at menopause. The ODF/OPG relative ratio was elevated at the concentration observed during ovulation in human gingival fibroblasts and at the concentration observed between ovulation and menstruation in periodontal ligament cells treated with estrogen. However, the ratio was <1 at all concentrations in both cells treated with progesterone. In the case of menopause simulated by estrogen depletion, the ratio was <1 in human gingival fibroblasts but >1 in periodontal ligament cells.

• Journal of Periodontal and Implant Science
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• v.26 no.3
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• pp.701-713
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• 1996

The purpose of this study was to evaluate the biocompatibility of the Nd:YAG lased root surface followed by root planing and/or tetracyline-HCI(T.C.-HCI) conditioning. $30,4mm{\times}4mm$ root segments were obtained from unerupted third molars and 21, periodontally involved root segments. The treatment groups were as follows : (1) healthy root cementum surface groups : 1) control(non-treated group), 2) lased only, 3) lased/root planed, and 4) lased/T.C.-HCI. (2) diseased root cementum surface groups : 1) control(root planed only), 2) lased/root planed, and 3) lased/root planed/T.C.-HCI. The specimens were treated with a Nd:YAG laser using a $320{\mu}m$ noncontact optic fiber handpiece with an energy setting of 1.5W($114.6J/cm^2$), 2.0W($152.9J/cm^2$), 5.0W($382J/cm^2$) for one minute. The fiber was held perpendicular to the petri dish(NUNC) 2cm apart in an attempt to expose the entire root segments equally. Human gingival fibroblasts were cultured from explants of normal interdental gingival tissue obtained during third morlar extraction. The attachment assay was performed with third-generation fibroblasts. The numbers of gingival fibroblasts attached to the root surface were counted on each specimen under the light microscope, and were statistically analyzed by the oneway ANOVA followed by Tukey's test in SPSS/PC+programs. The results were as follows : 1) In healthy root cementum surfaces, lased/root planed groups exhibited a significantly increased fibroblast attachment compared to controls, lased only, and lased/T.C.-HCI groups(p<0.05), 2) In diseased root cementum surfaces, laser treatment followed by root planing and/or T.C.HCl groups exhibited a increased tendency of fibroblast attachment compared to root planed only group. The results suggest that laser treatment followed by root planing and/or T.C.-HCl would appear necessary so as to render the root surface biocompatible.

• 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 Periodontal and Implant Science
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• v.31 no.3
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• pp.597-610
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• 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.

• International Journal of Oral Biology
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• v.44 no.4
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• pp.191-194
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• 2019

The purpose of this study was to investigate the antimicrobial effects of Australia propolis against cariogenic and periodontopathic bacteria. Antimicrobial activity was determined by evaluating the minimal bactericidal concentration (MBC). Cell cytotoxicity of propolis extract on normal human gingival fibroblast (HGF-1) cells was observed using the methylthiazolyldiphenyl-tetrazolium bromide assay. The data indicated that, with the exception of Aggregatibacter actinomycetemcomitans (KCOM 1306), the MBC values of the propolis strains were 0.25-1% without HGF-1 cell cytotoxicity. These results suggest that propolis can be used to develop oral hygiene products for the prevention of oral infectious disease.

• Journal of dental hygiene science
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• v.23 no.3
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• pp.216-224
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• 2023

Background: Smilax glabra has various pharmacological activities and is widely used as a herbal medicine. Although the incidence of oral cancer is low, the recurrence rate is high, and the 5-year survival rate is poor. It is necessary to search for anticancer drugs that increase the effect of cancer chemotherapy on heterogeneous oral tissues and reduce the side effects on normal cells. This study aimed to investigate the effects and mechanism of ethanol extract of Smilax glabra (EESG) as an anticancer drug for oral cancer. Methods: Smilax glabra root components extracted with 70% ethanol were used to analyze their effects on cancer cells. A 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide assay was performed for cytotoxicity analysis. Flow cytometry was performed to determine the cell cycle phase distribution. To observe apoptotic cells, terminal deoxynucleotidyl transferase dUTP nick end labeling and γH2AX were detected by fluorescence microscope. The protein levels of cleaved PARP and caspase were analyzed using western blotting. The activation of procaspase-3 was confirmed by measuring caspase-3 activity. Results: EESG was no cytotoxic to normal gingival fibroblast but was high in YD10B oral squamous cell carcinoma (OSCC) cells. EESG treatment increased the subdiploid DNA content of YD10B cells by assessing DNA content distribution. Chromatin condensation and DNA strand breaks increased in YD10B cells treated with EESG. EESG-treated YD10B cells had high Annexin V and low propidium iodide levels, confirming that early apoptosis was induced. In addition, increased levels of γH2AX foci, a marker of DNA damage, were observed in the nuclei of EESG-treated YD10B cells. The EESG-treated YD10B cells also exhibited decreased procaspase-3 and procaspase-9 levels, increased PARP cleavage and caspase-3 activity. Conclusion: These results indicate that EESG inhibited cancer cell proliferation by inducing apoptosis in YD10B OSCC cells.

• International Journal of Oral Biology
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• v.38 no.4
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• pp.141-147
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• 2013

It has been established that berberine has strong antimicrobial effects. Little is known however regarding the antimicrobial activity of berberine against endodontic pathogenic bacteria or its cytotoxicity in human oral tissue cells. The antibacterial properties of berberine were tested against 5 strains of Enterococcus faecalis and type strains of Aggregatibacter actinomycetemcomitans, Prevotella nigrescens, Prevotella intermedia, and Tannerella forsythia, which are involved in endodontic infections. Antimicrobial activity was evaluated through minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) measurements. The viability of normal human gingival fibroblast (NHGF) cells after exposure to berberine was measured using a methyl thiazolyl tetrazolium (MTT) assay. The data showed that berberine has antimicrobial effects against A. actinomycetemcomitans with an MIC and MBC of $12.5{\mu}g/ml$ and $25{\mu}g/ml$, respectively. In the cytotoxicity studies, cell viability was maintained at 66.1% following exposure to $31.3{\mu}g/ml$ berberine. Overall, these findings suggest that berberine has antimicrobial activity against the tested bacteria. Nevertheless, lower concentrations in combination with other reagents will need to be tested before these in vitro results can be translated to clinical use.