• Journal of Periodontal and Implant Science
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• 제36권3호
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• pp.757-765
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• 2006

Human periodontal ligament fibroblasts (hPDLF) are very important for curing the periodontal tissue because they can be differentiated into various cells. A tissue engineering approach using a cell-scaffold is essential for comprehending today's periodontal tissue regeneration procedure. This study examined the possibility of using an acellular dermal matrix as a scaffold for human periodontalligament fibroblast (hPDLF). The hPDLF was isolated from the middle third of the root of periodontally healthy teeth extracted for orthodontic reasons. The cells were cultured in a medium containing Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at $37^{\circ}C$ in humidified air with 5% $CO_2$. The acellular dermal matrix(ADM) was provided by the US tissue banks(USA). Second passage cells were used in this study. The hPDLF cells were cultured with the acellular dermal matrix for 2 days, and the dermal matrix cultured by the hPDLF was transferred to a new petri dish and used as the experimental group. The control group was cultured without the acellular dermal matrix, The control and experimental cells were cultured for six weeks. The hPDLF cultured on the acellular dermal matrix was observed by Transmission Electron microscopy (TEM). Electron micrography shows that the hPDLF was proliferated on the acellular dermal matrix. This study suggests that the acellular dermal matrix can be used as a scaffold for hPDLF.

• Journal of Periodontal and Implant Science
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• 제36권3호
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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.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2005년도 생물공학의 동향(XVI)
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• pp.224-224
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• 2005

• Restorative Dentistry and Endodontics
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• 제24권1호
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• pp.76-87
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• 1999

The effect of retrograde root-end filling materials(IRM, Super-EBA, Vitremer, MTA) on human periodontal ligament fibroblasts and osteoblasts was observed. The cell activities were evaluated by MTT assay, protein assay and alkaline phosphatase activity examination. The results as follows ; 1. After 24hrs culture, both E1 cells & PDL fibroblast adding root-end filling materials were suppressed cell activities but after 48hrs, cell activities were recovered. 2. Cell activity was lowest in Vitremer followed by IRM, MTA, Super-EBA. 3. Cell activity depression by Vitremer was not concerned with pH changes. 4. Protein synthesis by root-end filling materials were not significant difference in Both E1 cell & PDL fibroblasts but protein synthesis were a little increased by Super-EBA. 5. Alkaline phosphatase activity was increased in E1 cell by Super-EBA & MTA but was not significant differences in E1 cell by IRM & Vitremer. Alkaline phosphatase activity was a little depressed in PDL fibroblast by Vitremer. This findings suggest that these root-end filling materials may have important roles in promotion of PDL healing and consequently may be useful for clinical application in apical surgery.

• Restorative Dentistry and Endodontics
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• 제43권3호
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• pp.24.1-24.12
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• 2018

Objectives: The aim of this in vitro study was to evaluate the biocompatibility of newly proposed root-end filling materials, Biodentine, Micro-Mega mineral trioxide aggregate (MM-MTA), polymethylmethacrylate (PMMA) bone cement, and Smart Dentin Replacement (SDR), in comparison with contemporary root-end filling materials, intermediate restorative material (IRM), Dyract compomer, ProRoot MTA (PMTA), and Vitrebond, using human periodontal ligament (hPDL) fibroblasts. Materials and Methods: Ten discs from each material were fabricated in sterile Teflon molds and 24-hour eluates were obtained from each root-end filling material in cell culture media after 1- or 3-day setting. hPDL fibroblasts were plated at a density of $5{\times}10^3/well$, and were incubated for 24 hours with 1:1, 1:2, 1:4, and 1:8 dilutions of eluates. Cell viability was evaluated by XTT assay. Data was statistically analysed. Apoptotic/necrotic activity of PDL cells exposed to material eluates was established by flow cytometry. Results: The Vitrebond and IRM were significantly more cytotoxic than the other root-end filling materials (p < 0.05). Those cells exposed to the Biodentine and Dyract compomer eluates showed the highest survival rates (p < 0.05), while the PMTA, MM-MTA, SDR, and PMMA groups exhibited similar cell viabilities. Three-day samples were more cytotoxic than 1-day samples (p < 0.05). Eluates from the cements at 1:1 dilution were significantly more cytotoxic (p < 0.05). Vitrebond induced cell necrosis as indicated by flow cytometry. Conclusions: This in vitro study demonstrated that Biodentine and Compomer were more biocompatible than the other root-end filling materials. Vitrebond eluate caused necrotic cell death.

• 대한소아치과학회지
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• 제34권3호
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• pp.438-446
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• 2007

치주인대섬유모세포들은 완전탈구 된 치아의 성공적인 재식을 위한 중요한 요소이다. 따라서, 외상으로 인해 완전탈구된 치아의 보존을 위한 보관액을 선택하는 것이 중요하다. 성장인자들과 호르몬들은 치주인대섬유모세포들의 생존을 위한 치료적 제제로 고려되고 있다. Epidermal growth factor(EGF)는 다른 조직에서 재생과 상처 치유 과정의 중요한 역할인자로 대두되고 있다. 따라서, 완전탈구 된 치아를 위한 치료적 적용을 위해 EGF의 세포 증식에 미치는 영향을 평가하였다. 또한 EGF와 tri-iodothyronine(T3)의 혼합액, EGF와 Heparin-binding epidermal growth factor-like growth factor(HB-EGF)의 혼합액이 세포 증식에 미치는 상승 효과를 평가하였다. 치주인대섬유모세포의 세포증식은 EGF 농도가 증가함에 따라 증가하였고, 10 ng/ml 농도에서 최대 세포증식을 보였다. EGF는 상처치유분석에서 상처 치유촉진과 이동성을 보여주었다. EGF에 T3와 HB-EGF를 첨가한 혼합액에서 배양한 세포는 EGF만 처리한 경우보다 세포 증식이 상승되었다. EGF와 T3 혼합액의 상승효과 기전을 유추하기 위해서 RT-PCR로 EGF 수용기의 발현을 확인하였고, T3가 EGF 수용기 발현을 증가시켰음을 확인하였다. 따라서 EGF와, EGF와 T3, EGF와 HB-EGF의 혼합액은 완전탈구된 치아의 치료에 있어 유용한 선택이 될 것이다.

• Journal of Periodontal and Implant Science
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• 제41권1호
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• pp.17-22
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• 2011

Purpose: Nitric oxide (NO) has been known as an important regulator of osteoblasts and periodontal ligament cell activity. This study was performed to investigate the relationship between NO-mediated cell death of human periodontal ligament fibroblasts (PDLFs) and N-methyl-D-aspartic acid (NMDA) receptor antagonist (+)-5-methyl-10, 11-dihydro-5H-dibenzo[a,d]cyclohepten-5, 10-imine hydrogen maleate (MK801). Methods: Human PDLFs were treated with various concentrations (0 to 4 mM) of sodium nitroprusside (SNP) with or without $200\;{\mu}M$ MK801 in culture media for 16 hours and the cell medium was then removed and replaced by fresh medium containing MTS reagent for cell proliferation assay. Western blot analysis was performed to investigate the effects of SNP on the expression of Bax, cytochrome c, and caspase-3 proteins. The differences for each value among the sample groups were compared using analysis of variance with 95% confidence intervals. Results: In the case of SNP treatment, as a NO donor, cell viability was significantly decreased in a concentration-dependent manner. In addition, a synergistic effect was shown when both SNP and NMDA receptor antagonist was added to the medium. SNP treated PDLFs exhibited a round shape in culture conditions and were dramatically reduced in cell number. SNP treatment also increased levels of apoptotic marker protein, such as Bax and cytochrome c, and reduced caspase-3 in PDLFs. Mitogen-activated protein kinase signaling was activated by treatment of SNP and NMDA receptor antagonist. Conclusions: These results suggest that excessive production of NO may induce apoptosis and that NMDA receptor may modulate NO-induced apoptosis in PDLFs.

• Journal of Periodontal and Implant Science
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• 제30권4호
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• pp.869-885
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• 2000

Fibroblasts are major cellular components of gingiva and periodontal ligament. They regulate the healing process after surgery or injury. Recently, many natural medicines, whose advantages are less side effects and possibility of long-term use, have been studied for their capacity, their anti-bacterial and anti-inflammatory effects and regenerative potential of periodontal tissues. Sophorae radix have been traditionally used as an anti-bacterial and antiinflammatory drug in oriental medicine. The purpose of present study was to investigate the effects of Sophorae radix extract on cell cycle progression and its molecular mechanism in human gingival fibroblasts. Sophorae radix extracts($100{\mu}g/ml$) notably increased cell proliferation and cell activity in the human gingival fibroblasts as compared to non-supplemented controls. There was an increase in the S phase and a decrease in the G1 phase in $100{\mu}g/ml$ of Sophorae radix extracts group as compared to non-supplemented controls. The level of cyclin E and cdk 2 protein in test group was higher than that of control groups. But that of cyclin D, cdk 4, and cdk 6 was not distinguished from controls. The level of p53 protein in test group was lower than that of controls, whereas that of p21 was not different. The level of pRB protein in test group was higher than that of controls, whereas that of p16 was lower. These results indicate that the increase of cell proliferation by Sophorae radix extracts may be due to the increased expression of cyclin E and cdk 2, and the decreased expression of p53 and p16 in human gingival fibroblasts.

• 대한치과교정학회지
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• 제22권1호
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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.

• Restorative Dentistry and Endodontics
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• 제42권3호
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• pp.176-187
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• 2017

Objectives: This study aimed to evaluate the chemical and biological properties of fast-set white mineral trioxide aggregate (FS WMTA), which was WMTA combined with calcium chloride dihydrate ($CaCl_2{\cdot}2H_2O$), compared to that of WMTA. Materials and Methods: Surface morphology, elemental, and phase analysis were examined using scanning electron microscope (SEM), energy dispersive X-ray microanalysis (EDX), and X-ray diffraction (XRD), respectively. The cytotoxicity and cell attachment properties were evaluated on human periodontal ligament fibroblasts (HPLFs) using methyl-thiazoldiphenyltetrazolium (MTT) assay and under SEM after 24 and 72 hours, respectively. Results: Results showed that the addition of $CaCl_2{\cdot}2H_2O$ to WMTA affected the surface morphology and chemical composition. Although FS WMTA exhibited a non-cytotoxic profile, the cell viability values of this combination were lesser than WMTA, and the difference was significant in 7 out of 10 concentrations at the 2 time intervals (p < 0.05). HPLFs adhered over the surface of WMTA and at the interface, after 24 hours of incubation. After 72 hours, there were increased numbers of HPLFs with prominent cytoplasmic processes. Similar findings were observed with FS WMTA, but the cells were not as confluent as with WMTA. Conclusions: The addition of $CaCl_2{\cdot}2H_2O$ to WMTA affected its chemical properties. The favorable biological profile of FS WMTA towards HPLFs may have a potential impact on its clinical application for repair of perforation defects.