• Journal of Periodontal and Implant Science
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• 제26권2호
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• pp.414-428
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• 1996

The use of basic fibroblast growth factor which function as potent biologic mediators regulating numerous activities of wound healing has been suggested for the promotion of periodontal regeneration. The mitogenic effects of basic fibroblast growth factor on human periodontal ligament cells and human gingival fibroblasts were evaluated by determining the incorporation of 5-Bromo-2'deoxy-uridine into DNA of the cells in a dose -dependent manner. The cells which were prepared were the primary cultured gingival fibroblasts and periodontal ligament cells from human the fourth or sixth subpassages were used in the experiments. The cells which were seeded DMEM contain 10% FBS. The added concentrations of basic fibroblast growth factor were 0.1, 1, 10, 50, $l00{\eta}g/ml$ and basic fibroblast growth factor were added to the quiescent cells for 24 hours, 48 hours and 72 hours. They were labeled with $10{\mu}l/200{\mu}l$ 5Bromo-2'-deoxy-uridine for the last 6 hours of each culture. The results of the five determinants were presented as mean and S.D.. The results were as follows. : The DNA synthetic activity of human gingival fibroblasts was increased dose dependently by basic fibroblast growth factor at 24 hours, 48 hours and 72 hours. The similar mitogenic effects were at the 24 and 48 hours of basic fibroblast growth factor, but the DNA synthetic activity of human gingival fibroblasts generally decreased at 72 hours. The DNA synthetic activity of human periodontal ligament cells was increased dose dependently to $50{\eta}g/ml$ by basic fibroblast growth factor at 24, 48 and 72 hours, but the DNA synthetic activity decreased at $l00{\eta}g/ml$ of each hour. Generally the maximum mitogenic effects were at the 48 hours application of basic fibroblast growth factor. The DNA synthetic activity of human periodontal ligament cells generally decreased lower at 72 hours than at 24, 48 hours the application of basic fibroblast growth factor. In the comparison of DNA synthetic activity between human gingival fibroblasts and human periodontal ligament cells, human periodontal ligament cells had slightly higher proliferation activity than human gingival fibroblasts for a longer time at the high dosage of the basic fibroblast growth factor.In conclusion, basic fibroblast growth factor have important roles in the stimulation of DNA synthesis in human periodontal ligament cells and human gingival fibroblasts, and thus may be useful for clinical applications in periodontal regenerative procedures.

• Archives of Plastic Surgery
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• 제34권4호
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• pp.420-425
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• 2007

Purpose:The mechanism of scar formation is not fully understood. Fibroblast is an important cell in wound healing process. We experienced a patient who was taking progesterone orally. Upper blepharoplasty was performed on her but, wound healing was delayed. We hypothesized that progesterone was the cause of delayed wound healing and fibroblast proliferation inhibition. We investigated the effect of progesterone in vitro on human dermal fibroblasts to study the effects on fibroblast proliferation. Methods: Human dermal fibroblasts from four persons were cultured initially. Progesterone is mixed to them at various concentrations, and fibroblast cell count was measured by MTT assay method at 570 nm. We confirmed that progesterone has some inhibitory effect on fibroblast proliferation and maximal inhibitory concentration of progesterone was determined. Then fibroblasts from a total of nineteen persons were cultured and the effects of progesterone were studied. Results: The initial study showed the maximal inhibitory concentration of progesterone to be $50{\mu}g/ml$. The main study showed that progesterone had 70.9% inhibitory effect on human dermal fibroblast in vitro. Conclusion: Progesterone has inhibitory effect on cultured human dermal fibroblast proliferation in vitro.

• Archives of Plastic Surgery
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• 제32권3호
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• pp.347-356
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• 2005

The wound healing process in fetus is quite different form that of adult. Regeneration plays an important role and scarless wound healing is possible in early gestational fetal period. Recently, the various effects of the hypoxia and reoxygenation in the wound healing process have been investigated by many researchers. The hypoxic state is known to alter protein synthesis and gene expression of TGF-${\beta}$, VEGF. The authors hypothesize there may be differences between fetal and adult fibroblast and this difference may play a possible role in the mechanism of scarless fetal wound healing. In this study, we investigated the growth of fibroblast, the amount of collagen deposition, the amount of protein synthesis and gene expression in TGF-${\beta}$(transforming growth factor-${\beta}$), VEGF(vascular endothelial growth factor) under the various hypoxic and reoxygenation conditions. Through these processes, we tried to determine the relationships between scarless fetal wound healing and hypoxic condition. In control group, fetal and adult fibroblasts were cultured under normoxic condition. The experimental groups were allocated into four different groups. The differences in TGF-beta, VEGF under 24, 48, 72 hours were statistically investigated. Compared to adult fibroblast group, there was a statistically significant increase (p<0.01) in the rates of protein synthesis in TGF-beta and VEGF of fetal fibroblast. In this study, these results may reflect the possibility that fetal fibroblast are more susceptible to change in oxygen and has a superior rate of angiogenesis through increased VEGF expression. The possible superiority of angiogenesis in fetal fibroblast may play an important role in scarless wound healing.

• Journal of Periodontal and Implant Science
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• 제25권1호
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• pp.133-145
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• 1995

The migration and proliferation of periodontal ligament cells are desired goal of periodontal regeneration therapy. PDGF and $TGF-{\beta}1$ are well known to regulate the cell activity of mesenchymal origin cell. The purpose of this study was to determine the effects of these growth factors on human gingival fibroblast and periodontal ligament cell actvity, and to identify the regulatory effect of $TGF-{\beta}1$ on the response to PDGF by MIT assay. Human gingival fibroblast and periodontal ligament cells were cultured from extracted teeth for non-periodontal reason. Cultured human gingival fibroblast and periodontal ligament cells in vitro were treated with polyperpetide growth factor PDGF and $TGF-{\beta}1$ in both a dose and time - dependent manner. Cell morphology were determined by inverted microscope and cell acitivity were determined by MIT assay. The result of this study demonstrated that PDGF and $TGF-{\beta}1$ were not changed the morphology of these cell compared with control group. PDGF or $TGF-{\beta}1$ increased cell activity of periodontal ligament cell in dose and time dependent manner but gingival fibroblast were decreased to the level of control group at third day. Additionally, incubation with $TGF-{\beta}1$ addition to PDGF resulted in a enhanced cell activity of PDGF. Therefore, cell acitivty of gingival fibroblast were not changed compared with control group. This stiudy demonstrates that PDGF and $TGF-{\beta}1$ are major mitogens for human periodontal ligament cell in vitro, and $TGF-{\beta}1$ is a regulator of cell activity to PDGF in human gingival fibroblast and periodontal ligament cell.

• Journal of Periodontal and Implant Science
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• 제26권4호
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• pp.889-905
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• 1996

It is known that growth factors function as potent biologic mediators regulating numerous activities of wound healing via cell proliferation, migration and extracellular matrix formation and they also promote periodontal regeneration. But, method of growth factor application is controversial yet. So purpose of this study is to evaluate the effect of demineralized root surface as one of method of growth factor application. The ginigival fibroblasts were primary cultured and fifth or sixth subpassages were used in these experiments. In first experiment, root surface blocks demineralized with 100mg/ml tetracycline for 5 minutes and pH 1 citric acid for 3 minutes(experimental groups) and nonteminerilized root surface blocks (control groups) were placed in 100ng/ml PDGF-BB for 5 minutes. Then the cells were seeded on each root surface blocks and cultured for 6, 24, 48, 72 hours. In second experiment, root surface blocks deminerilized with tetracycline and citric acid and nondemineralized root surface blocks were placed in 200ng/ml PDGF-BB for 5 minutes and another non-demineralized root surfcae blocks were placed in DMEM without PDGF-BB. At 1, 2, 4, 6, 8 days, the cells were seeded in 24-well plate and using of each eluent, cultured for 72 hours. The results of the four determinants were presented as mean and S.D.. The results were as follows : The attachment and proliferation of human gingival fibroblast on root surface were more increased when PDGF-BB was applicated on root surfrace demineralized with tetracycline or citric acid than non-demineralized root surface. And, in comparision tetracycline with citric acid, there were more attachment and proliferation of human gingival fibroblast on root surface demineralized with tetracycline than citric acid, and proliferation of human gingival fibroblast on demineralized root surface was increased time dependently 1 day to 3 days. In second experiment using eluent, proliferation of human gingival fibroblast was more increased to 6 days when human gingival fibroblast was cultured in eluent that PDGF-BB was applicated on demineralized root surface than two control groups, and degree of proliferation was decreased time dependently 1 day to 6 days. Proliferation of human gingival fibroblast cultured in eluent without PDGF-BB was constant 1 day to 6 days. After 6 days, degree of proliferation of human gingival fibroblast was similar in four groups. This means that release duration of PDGF-BB from demineralized root surface is 6 days. And in comparision tetracycline with citric acid, there was more proliferation of human gingival fibroblast in tetracycline-treated group than citric acid. In conclusion, demineralized root surface as primary site for PDGF-BB application, especially demineralized with tetracycline has important roles in attachment and proliferation of human gingival fibroblast, and may be useful clinical applications in periodontal regenerative procedures.

• Biotechnology and Bioprocess Engineering:BBE
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• 제3권2호
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• pp.51-54
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• 1998

It has been proved that co-cultivation of human neroblastoma cells and human fibroblast cells can enhance nerve cell growth and the production of BDNF in perfusion cultivation. In batch co-cultivation, maximum cell density was increased up to 1.76${\times}$106 viable cells/mL from 9${\times}$105 viable cells/mL of only neuroblastoma cell culture. The growth of neuroblastoma cells was greatly improved by culturing both nerve and fibroblast cells in a perfusion process, maintaining 1.5${\times}$106 viable cells/mL, which was much higher than that form fed-batch cultivation. The nerve cell growth was greatly enhance in both fed-batch and perfusion cultivations while the growth of fibroblast cells was not. It strongly implies that the factors secreted from human fibrobast cells and/or the environments of co-culture system can enhance both cell growth and BDNF secretion. Specific BDNF production rate was not enhanced in co-cultures; however, the production period was increased as the cell growth was lengthened in the co-culture case. Competitive growth between nerve cells and fibroblast cells was not observed in all cases, showing no changes of fibroblast cell growth and only enhancement of the neuroblastoma cell growth and overall BDNF production. It was also found that the perfusion cultivation was the most appropriate process for cultivating two cell lines simultaneously in a bioreactor.

• 대한치과보철학회지
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• 제44권1호
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• pp.112-123
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• 2006

Purpose: The biocompatibility and bio-adhesive property of a dental implant abutment are important for proper soft tissue healing and maintenance of osseointegration of implant. However, studies of soft tissue healing and mucosal attachment of various materials of implant abutment other than titanium are still needed. In this study, cell attachment, proliferation, cytotoxicity of human gingival fibroblast for ceramic, gold alloy, Ni-Cr alloy and, commercially available pure titanium as a control were evaluated, using MTS and scanning electron microscopy. Materials and Methods: Specimen was designed to disc, 4mm diameter and 1mm thickness, made of ceramic, gold alloy, Ni-Cr alloy and commercially available pure titanium. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. Cells were inoculated in the multiwell plates placed the specimen disc. Cell Titer 96 AQucous One Solution Cell Proliferation Assay were done after 1hour 3hours, 24hours, 3days, 5days of incubation. The discs were processed for scanning electron micrography to evaluate cell attachment and morphologic change. Results: The results were obtained as fellows. 1. The ceramic showed high cell attachment and proliferation and low cytotoxicity, which is as much bioadhesive and biocompatible as titanium. 2. The gold alloy represented limited proliferation of human gingival fibroblast and the highest cytotoxicity among tested materials (p<0.05). 3. The Ni-Cr alloy limited the proliferaion of the human gingival fibroblast compared to titanium(p<0.05) but cytotoxicity on the bottom of well was not so considerable, compared to titanium. 4. On the scanning electron micrographs , the ceramic showed good attachment and proliferation of human gingival fibroblast, which was similar to titanium. But gold alloy and Ni-Cr alloy showed the shrinkage of gingival fibroblast both after 24 hours and 3 days. On 5th day, small amount of the human gingival fibroblast proliferation was observed on the Ni-Cr alloy, while the shrinkage of gingival fibroblast was still observed on the gold alloy. Conclusions: These results suggest that the ceramic abutment is as biocompatible as titanium to make proper mucosal seal. The gold alloy has a high cytotoxicity to limit proliferation of gingival fibroblast, which suggest limited use on the anterior tooth where soft tissue healing is recommeded.

• Journal of Periodontal and Implant Science
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• 제25권1호
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• pp.1-13
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• 1995

Chlorhexidine and Listerine are widely used in dentistry due to its effectiveness on plaque control and bactericidal action. The effects of these agent on chronic gingivitis and wound healing following surgical periodontal therapy in human has been favorable. Understanding the effects of chlorhexidine and Listerine on human gingival fibroblast will provide the rationale for its use during the healing process of periodontal surgery. The purpose of this study was to compare the effects of chlorhexidine and Listerine on human gingival fibroblast. Human gingival fibroblasts were cultured from the healthy gingiva on the extracted premolar of orthodontic patients. Human gingival fibroblast were trypsinized and cultured in growth medium added range of 0.0012-0.12% chlorhexidine and 1-100% Listerine mouth wash solution. The cell used in this study were between fifth to eighth passage number. The cell morphology were examined by inverted microscope and the cell activity were measured by MIT assay. The Morphology of gingival fibroblast added Chlorhexidine and Listerine at the concentration of all range were became globular and lost their cytoplasmic process. Our results indicate that a 0.0012 concentration of chlorhexidine and 1% concentration of Listerine were shows minimal cytotoxicity, but above these concentraion, there was a significant difference between the cell activity in the experimental group and control group(p

• Medical Lasers
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• 제9권1호
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• pp.12-24
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• 2020

Human fibroblast-derived multi-peptide factors (MPFs) have been used during treatments with energy-delivering modalities to enhance energy-induced tissue reactions. Human fibroblast-derived MPFs, which include a range of growth factors and chemoattractive factors, activate and recruit fibroblasts and endothelial cells, as well as promote extracellular matrix deposition, all of which are crucial to wound repair. Interestingly, fibroblasts from different species or anatomical sites exhibit distinct transcriptional properties with high heterogeneity. In addition, the patterns of MPF secretion can differ under a range of experimental conditions. Therefore, the use of allogeneic fibroblasts and proper cultivation thereof are necessary to obtain MPFs that can enhance the epithelial-mesenchymal interactions during wound repair. Moreover, energy-delivering devices should be selected according to evidence demonstrating their therapeutic efficacy and safety on a pathological skin condition and the major target skin layers. This paper reviewed the histologic patterns of post-treatment tissue reactions elicited by several energy sources, including non-ablative and ablative fractional lasers, intense focused ultrasound, non-invasive and invasive radiofrequency, picosecond-domain lasers, and argon and nitrogen plasma. The possible role of the immediate application of human fibroblast-derived MPFs during wound repair was proposed.

• 한국식품영양과학회지
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• 제26권6호
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• pp.998-1005
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• 1997

Kimchi showed protective effect from oxidative damage generated by hydrogen peroxide and paraquat. To investigate the major components of kimchi which reduce the cytotoxicity against reactive oxygen species, keratinocyte(A431, epidermoid carcinoma, human) and fibroblast(CCD-986SK, normal control, human) were cultured under oxidative stress condition provoked by paraquat, a superoxide anion generator, and hydrogen peroxide in the absence or presence of kimchi ingredients. Most keratinocyte and fibroblast cells were killed by hydrogen peroxide and paraquat over 1mM concentration, but kimchi ingredients showed protective effects from oxidative damage generated by hydrogen peroxide and onion, among those, garlic showed the most remarkable preventive effect. Most of kimchi ingredients showed protective effect against paraquat, especially leek notably increased cell survival. For fibroblast cells, ginger had the preventive effect against paraquat, especially leek notably increased cell survival. For fibroblast cells, ginger had the preventive effect from cell killing by high dose of hydrogen peroxide, but most ingredients were not effective against paraquat.