• Restorative Dentistry and Endodontics
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• 제18권1호
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• pp.156-172
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• 1993

The purpose of this study was to evaluate for the cytotoxicity of root canal sealers - Tubliseal, N2, AH26, and Sealapex - on fibroblasts cultivated from human periodontal ligament. Succinate dehydrogenase activity test and $^{51}Cr$ release test were performed to evaluate the shortterm cytotoxicity. According to l,3 and 6 days vital cell count and $^{14}C$-leucine incorporation rate to fibroblasts were evaluated. The results of this study were as follows: 1. In the test of SDH activity by millipore filter method, Sealapex showed mild cytotoxicity but Tubliseal, N2 and AH26 showed severe cytotoxicity. 2. In $^{51}Cr$ release test, Tubliseal was the most cytotoxic sealer tested, and rank ordered the relative cytotoxicity of the other sealers tested as follows: AH26, N2, Sealapex. 3. In the test of viable cell count, cytotoxicity in Tubliseal was continued because vital cell number reduced with time. Because vital cell was not showed in N2 and AH26 at 1 day, it was recognized that N2 and AH26 exhibited severe cytotoxicity. In Sealapex, vital cell number increased remarkably with time, so it showed that cytotoxicity decreased with time. 4. In $^{14}C$-leucine incorporation rate test, protein sythesis was not produced in Tubliseal, N2 and AH26 after 3 days, it showed that cytotoxicity in Tubliseal, N2 and AH26 was severe. Although protein synthesis in Sealapex decreased with time, it continued after 6 days. Therefore Sealapex has been exhibited mild cytotoxicity.

• Journal of Periodontal and Implant Science
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• 제35권1호
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• pp.21-30
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• 2005

Matrix metalloproteinases (MMPs) are a family of host-derived proteolytic enzymes and implicated in the remodeling and degradation of extracellular matrix under both physiological and pathological conditions. Connective tissue degradation in periodontal diseases is thought to be due to excessive MMP activities over their specific inhibitors. The effects of lipopolysaccharide (LPS) from Prevotella intermedia, one of the major putative pathogens of periodontitis, on the expression of mRNA for MMPs and tissue inhibitors of metalloproteinases (TIMPs) in human gingival and periodontal ligament fibroblasts were examined by reverse transcriptase-polymerase chain reaction (RT-PCR). The expression of mRNAs encoding MMP-1, -2, -3, -10, and -14 was increased in human gingival fibroblasts treated with p. intermedia LPS, whereas MMP-11 and TIMP-2 mRNA expression was decreased in these cells stimulated with LPS. P. intermedia LPS increased the MMP-1, -2, -10, -11, and -14 mRNA expression and decreased TIMP-1 and -2 mRNA expression in human periodontal ligament fibroblasts. These findings imply that P. intermedia LPS may play an important role in the connective tissue degradation in periodontitis.

• Journal of Periodontal and Implant Science
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• 제25권2호
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• pp.210-221
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• 1995

The purpose of this study was performed to investigate the mineralization and differentiation of osteobalsts for bone regeneration in vitro and the effect of rate of the composition in periodontal cells on mineralization. For this study, healthy gingival tissues were surgically obtained from the patients during 1st premolar extraction for the purposes of orthodontic treament. Gingival tissue was washed several time with Phosphate buffered saline contained high concentration of antibiotics and antifungal agent, and cultured in Dulbecco's Modified Eagle's Medium(DMEM, Gibco, U.S.A.). Every cell were cultured in state at $37^{\circ}C$, 100% of humidity, 5% of $CO_2$ incubator. Bone marrow stromal cells were isolated from 5-clay-old rat femur with using medium irrigation mathod by syringe. Cell suspension medium were centrifuged at 1500 rpm for 5 min and then cultured in the petri dish. Two kinds of cell were freezed and stocked in the liquid nitrogen tank until experiment. Cell were incubated into the 24 multi-well plate with $5{\times}10^4$cell/well of medium at $37^{\circ}C$, 100% of humidity 5% $CO_2$ incubator for 24 hours. After discarded of the supernatent of medium, O.5ml of medium were reapplied and incubated. And counted the number of cell using the hemocytometer and inverted light microscope. We have measured the number of mineralized nodule with using Alizarin red S. staining in microscope. Furthermore every cell were observed the morphological change between every rate of co-culture of the two kinds of cell. The results were as follows; The rate of proliferation of co-culture cell revealed high rate tendency compared the bone marrow stromal cell only and low growth rate to compared with gingival fibroblast only. The tendency of formation of the mineralized nodule were observed dose-depend pattern of bone marrow stromal cell. It is concluded that the gingival fibroblast may inhibit the formation of mineralized nodule in the culture of the bone marrow stromal cell.

• 대한치과교정학회지
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• 제30권3호
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• pp.317-333
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• 2000

급속 구개확장은 정중구개봉합을 이개하기 위하여 강한 힘을 치아에 가하게 된다. 확장후 정중구개봉합부에 존재하는 골모세포와 섬유모세포의 증식 활성도를 측정하여 정중구개확장이 생리적으로 진행되는지를 확인하고, 이때 개개 치아의 치주조직에서 나타나는 변화를 조사하기 위하여 약 10개월된 유성견 10마리를 대상으로 상악 제2소구치와 제1대구치에 Hyrak screw를 장착하였다. 실험동물은 대조군 1마리, 1주 확장한 군 3마리, 2주 확장한 군 3마리, 2주 확장후 2주 보정한 군 3마리로 구분하여, Screw를 1일 1/4회전시켜 확장하였고, 제2소구치, 제3소구치, 그리고 제 1대구치 부위의 정중구개봉합부와 치주조직을 절취하여 Soft X-ray로 관찰하고, 형태학적 및 면역조직화학적 변화를 광학현미경으로, 그리고 골개조를 편광현미경으로 관찰하여 다음과 같은 결과를 얻었다. 1. 편광현미경 소견에서는 대조군에 비하여 1주 확장군의 봉합부에서 골 형성과 흡수가 크게 증가되었으며, 2주 확장군에서 약간 감소를 보이다가, 보정군에서는 석회화가 크게 진행되었다. 2. 면역조직화학적 연구에서는 대조군에 비하여 1주 확장군의 봉합부에서 골모세포가 PCNA(Proliferating Cell Nuclear Antigen)에, 미분화 섬유모세포가 EGF(Epidermal Growth Factor)에 약간 증가된 양성 반응을 보였으며 EGF에 대한 골모세포의 반응은 낮았다. 2주 확장군에서는 섬유아세포와 골모세포에서 PCNA와 EGF에 대한 양성반응이 증가되었다. 보정군에서는 이들 PCNA와 EGF에 양성반응을 나타내는 세포들이 구개돌기 골단 주변에 집중되었다. 3. 확장기간 중 봉합부 주변에서 광범위한 골 흡수와 골형성을 동반한 골개조가 일어났으며, 보정군에서는 다소 감소하였다. 2주 확장군과 보정군의 제3소구치 와 제1대구치의 치근단에서 백악질 형성과 흡수가 나타났으며, 제1대구치의 협측 치경부에서 광범위한 초자양 변성대가 나타났다. 4. Soft X-ray소견에서 1주 확장군은 대조군에 비교하여 봉합부위의 여러부분에서 미세 파절과 저석회화된 결손부위를 보였다. 대조군과 보정군 사이 에는 정중구개봉합부의 석회화 정도에 별 차이 가 없었다. 2주 확장군과 보정군에서 제3소구치와 제1대구치는 협측으로 경사되는 경향을 보였다.

• Journal of Periodontal and Implant Science
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• 제30권3호
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• pp.599-610
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• 2000

Gingival fibroblasts are embedded in an extracellular matrix. The matrixs have influence on the development, polarity, and behavior of nearby cells. The major component of periodontal extracellular matrix is a glycosaminoglycan. The glycosaminoglycan are large carbohydrates that are composed of repeating disaccharide units and exist in three main form: dermatan sulfate, chondrotitin sulfate, heparan sulfate. The purpose of present study is to examine the biologic effects of glycosaminoglycan on human gingival fibroblast. Human gingival fibroblasts were supplemented with each glycosaminoglycan, and cellular attachment and proliferation was determined by MTT assay. Dermatan sulfate and chondroitin sulfate did not stimulate the attachment and proliferation of human gingival fibroblasts, but heparan sulfate increased the proliferation and attachment in a time- and dose dependent manner. These results indicated that heparan sulfate seems to have a high potential for gingival regeneration and root surface attachment.

• Journal of Periodontal and Implant Science
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• 제24권3호
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• pp.661-670
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• 1994

It is well known that the application of dressings after periodontal surgery have benefits to provide the comforts to patient and to promote the healing process with action of bleeding control and temporary stabilization for the operated mobile teeth. But until recently the relationship between periodontal dressings and cells which are composed of periodontium has not been clear. The purpose of this study was to evaluate the cytotoxic effect of soluble extracts from the four different kinds of periodontal dressings, two of them were eugenol type (K.H.pack, Wondrpak) and the others were non-eugenol type (Coe-pak, Periocare), on the human gingival fibroblasts in vitro. Human gingival fibroblasts were primarily cultured from gingiva around third molar during the extraction for preventive purposes. Extracts solution were prepared with culture medium by means of imersing the consistent size of periodontal dressing made from plastic mold. Cell were inoculated into the 24 well plate with $3\;{\times}\;10^4\;cells/well$ of medium at $37\;^{\circ}C$, 100% of humidity, 5% of $CO_2$, incubator for 24 hours. After discard of the supernatant of medium, those cells were cultured with original, 1/2, 1/5, 1/10 diluted soluble extract for 24, 48 and 72 hours, and counted the number of cells using the hemocytometer at each designed time and concentration. Also, the cytotoxic effect of soluble extract was measured by Wataha's MTT assay method. In briefly, cells were inoculated and cultured into 96 well culture plate with $2\;{\times}\;10^4\;cells/well$ for 24 hours. Soluble extracts were applied to cultured cells and incubated for 48 hours at same condition. $50\;{\mu}l$ of MTT solution and DMSO were added into each well for the detection of absorbance with ELISA reader. The measured data were calculated by value of colorimetric assay for survival rate. The results were as follows ; In the case of eugenol type of dressing, original, 1/2 and 1/5 diluted extracts of K.H.pack showed very low survival rate. And original extract of Wondrpak showed strong cytotoxic effect and 1/2 diluted extract showed moderate cytotoxic effect. In the case of Non-eugenol type of dressings, only original extract of Coe-pak revealed strong cytotoxic effect and Periocare had little cytotoxic effect. It is concluded that eugenol type of dressings showed more cytotoxic effect than non-eugenol types. This study suggest that use of non-eugenol dressings after periodontal surgery is recommended.

• Journal of Periodontal and Implant Science
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• 제26권1호
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• pp.89-102
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• 1996

The goal of periodontal therapy is to regenerate the loss of periodontal attachment appratus. Current theories suggest the cells of the periodontium have the capacity, when appropriately triggered, to actively participate in restoring connective tissues, including mineralized tissues. This study was performed to define the hard tissue regeneration effect of periodontal ligament(PDL) cells in vitro and the effect of rate of the composition in gingival fibroblasts(GF) on the hard tissue regeneration capacity of PDL cells. For this study, Cell growth rate, alkaline phosphatase(Al.Pase) levels and the ability to produce mineralized nodules in co-culture of PDL cells and GF were examined. The results were as follows : 1. At 7 and 15 days, Cell growth of co-culture of PDL and GF(50 : 50) was greater than that of PDL cells or GF alone(P>0.05). 2. Measurements of ALPase levels indicated that PDL cells had significantly higher activity when compared with that of co-culture groups and GF only(p<0.05). And, ALPase activity in 10 days was higher than that of 7 days(P>0.05) 3. The tendency of formation of the mineralized nodule were observed dose-depend pattern of PDL cells. There was statistically significant difference among group 1(PDL 100%), 2(PDL 70% : GF 30%), and 3(PDL 50% : GF 50%)(P<0.01). But, there was no difference among group 3, 4(PDL 30% GF 70%), and 5(GF 100%). 4. Also, the number of nodule was greater in co-culture of PDL 70% and GF 30% than in culture of PDL 70%(P<0.05) From the above results, it is assumed that the co-culture of PDL cells and GF stimulates the cell growth, which is not that of PDL cells but GF. And, the activity of ALPase depends on the ratio of PDL cells, and ALPase may relate to the initial phase of nodule formation. Also, it is thought that the calcified nodule formation principally depends on PDL cells, is inhibited by GF, and affected by cell density.

• Journal of Periodontal and Implant Science
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• 제30권4호
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• pp.895-913
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• 2000

It is becoming increasingly clear that human gingival fibroblasts(HGF) may play a role in regulating immune responsiveness in inflammatory periodontal lesions. Stimulation of HGF with locally-secreted T cell cytokine $IFN_{\gamma}$ induces human leukocyte antigen class II(HLA II) expression on HGF, which is one of the characteristic feature of professional antigen presenting cells(pAPC). However, $IFN_{\gamma}$-treated HGF and other nonprofessional antigen presenting cells(npAPC) are known to be ineffective or less effective antigen presenter to resting T cells. This study, therefore, was undertaken in an effort to elucidate the differences in expression of cell surface molecules between npAPC in periodontal tissues, such as HGF and periodontal ligament fibroblasts(PDLF), and pAPC such as monocytes/macrophages. Using flow cytometry, the levels of cell surface expression of HLA-D, ICAM-1, LFA-3, and B7-1, which are involved in antigen presentation, were determined in HGF, PDLF and human myelomonocytic cell line THP-1. $IFN_{\gamma}$ clearly induced HLA-D expression on both of fibroblasts and monocytes dose dependently. However, expression level on monocytes were 4 to 5 times higher than that on fibroblasts, and induction rate was faster in monocytes than in fibroblasts. The levels of ICAM-1 expression on fibroblasts and monocytes were enhanced by $IFN_{\gamma}$ in a dose dependent manner. On the other hand, the expression of LFA-3 molecule, which could be detected in fibroblasts and monocytes without cytokine stimulation, was no more enhanced by addition of $IFN_{\gamma}$. B7-1, important costimulatory molecule in T cell activation and proliferation, was not detected on both of fibroblasts and monocytes even when stimulated with $IFN_{\gamma}$, except on monocytes fully differentiated by pretreatment of PMA and treated by $IFN_{\gamma}$. These results suggest that delayed expression of HLA-D and absence of B7-1 on $IFN_{\gamma}$ - treated fibroblasts may at least in part be involved in the ineffectiveness of fibroblasts as primary APC. And it is postulated that although periodontal fibroblasts may not serve as primary APC in normal periodontium, sustained expression of HLA II on ubiquitous fibroblasts in inflammatory lesions may perpetuate immune responses and produce chronic inflammation and tissue injury.

• Journal of Periodontal and Implant Science
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• 제38권4호
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• pp.679-690
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• 2008

Purpose: The aim of this study was to investigate the effects of EMD on demineralized root surface using human periodontal ligament cells and compare the effects of root conditioning materials(tetracycline(TCN), EDTA). Material and Methods: Dentin slices were prepared from the extracted teeth and demineralized with TCN and EDTA. Demineralized dentin slices were incubated at culture plate with 25, 50 and $100{\mu}g/ml$ concentration of EMD. Cell attachment, alkaline phosphatase activity test, protein synthesis assay and scanning electronic microscopic examination were done. Results: Cells were attached significantly higher in EMD treated group at 7 and 14 days. Cell numbers were significantly higher in EMD treated group. Alkaline phosphatase activity was significantly higher in EMD treated group at 7 and 14 days. Protein synthesis was significantly higher in EMD treated group at 7 and 14 days. Conclusion: Enamel matrix derivatives enhance the biologic activities of human periodontal ligament cells on demineralized root surface and its effects are dependent on the concentration of EMD.

• Journal of Periodontal and Implant Science
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• 제32권1호
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• pp.249-255
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• 2002

Zea Mays L. has been known to be effective for improving tissue health and Magnoliae cortex to have effective antibacterial and antimicrobial activity against pathogenic microbes. The purpose of this study was to examine the antimicrobial effects of Zea Mays L. and Magnoliae cortex extract mixtures on periodontal pathogens(Prevotella intermedia, Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Streptococcus mutans )and to examine the effects on human gingival fibroblast cellular activity. Zea Mays L. and Magnoliae cortex extracts and their mixtures were prepared with various mixing ratios (0.5:1, 1:1, 1.5:1, 2:1). These extracts were loaded to periodontal pathogen cultured petri dish for antimicrobial test and also loaded to cultured human gingival fibroblast for cellular activity test. Each test was repeated 3 times and data were analyzed by one-way ANOVA with 95% confidence level. Mixture of these two extracts showed greater amount of inhibition area on periodontal pathogen and more improved gingival fibroblast activity as Zea Mays L. ratio reduced. So, mixture ratio 0.5:1 (Zea Mays L. : Magnoliae cortex) group showed statistical significance in antimicrobial activity and cellular activity among various mixtures(p < 0.05). In conclusion, 0.5:1 (Zea Mays L. : Magnoliae cortex) mixture possessed best gingival fibroblast cellular activity and antimicrobial activity toward periodontal pathogens.