• 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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• 제28권4호
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• pp.677-692
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• 1998

In order to study the effects of cigarette smoking on periodontal tissue, gingival fibroblast from the smoking and nonsmoking groups were cultured and each group were treated with nicotine(50ng/ml,100ng/ml) and NNK(50ng/ml, 100ng/ml) to test their attachment ability at time intervals of 30minutes, 60minutes, 90minutes, 120minutes, and 240minutes. Using the same method, the growth each group treated with nicotine and NNK in order to compare their attachment ability and growth rate was done. The Results are as follows. 1. In comparing the attachment ability and growth rate between the smoking and non-smoking group were significantly higher in all time intervals. 2. When the attachment ability was com-pared among these two groups after treatment with nicotine and NNK, the non-smoking group showed decrease in attachment ability while the smoking group was not affected. 3. The growth rate of these two groups were compared after treating with nicotine and NNK. The growth rate of fibroblast from the non-smoking group decreased while fibroblast from the smoking group was not affected. These results suggest that fibroblast from the non-smoking group showed higher attachment ability, growth rate, and sensitivity to nicotine and NNK. This implies that fibroblast from the non-smoking group is a more reliable source in testing the cytotoxicity of nicotine and NNK. Also it could be reasonable to think that nicotine and NNK is a probable cause for problems in attachment and repair mechanism.

• Archives of Plastic Surgery
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• 제35권5호
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• pp.501-506
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• 2008

Purpose: In order to overcome the limitations of the conventional cryopreserved fibroblast or keratinocyte allograft method used in the treatment of diabetic foot ulcers, we reported a pilot study in 2004 demonstrating promising results of a fresh fibroblast allograft method in eight patients. However, the number of cases was insufficient for full evaluation and the follow-up duration was not long enough to determine the efficacy and safety of the method. This encouraged us to conduct this follow-up study to fully evaluate the use of noncryopreserved fresh human fibroblast allografts in treating diabetic foot ulcers. Methods: Thirty-seven patients with diabetic foot ulcers were treated using fresh fibroblast allografts. Human dermal fibroblasts from healthy teenagers were cultured in DMEM/F-12 medium supplemented with 10% serum. The cultured cells were applied on the wounds immediately following debridement, with fibrin being used as a cell carrier. In eight weeks, percentages of complete healing, mean healing time, and patient satisfactions were assessed, with follow-up time ranging from 6 to 40 months. Results: Our study showed that 83.8% of the treated patients were complete healed. The time required for complete healing was $30.9{\pm}10.1$ days. Patient satisfaction scores for the experimental treatment were higher than those for the conventional method(mean scores of $8.1{\pm}1.1$ and $4.8{\pm}1.4$, respectively). No adverse events related to the study treatment occurred. Conclusion: The use of fresh human fibroblast allografts was found to be a safe and effective treatment for diabetic foot ulcers.

• 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.

• Journal of Periodontal and Implant Science
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• 제25권2호
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• pp.222-238
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• 1995

Inflammatory cells may produce active species of oxygen in antimicrobial defense. While such species can directly damage surrounding tissue, their major secondary role may be to mediate important components of the inflammatory response. Superoxide dismutase, antioxidant, have significant anti-inflammatory properties in rheumatoid arthritis, ischemic tissue injury and gastrointestinal disease. Increased oxidative product formation diseases. And superoxide dismutase produced by Porphyromonas Gingivalis is resistant to killing by polymorphonuclear leukocyte. The purpose of this study was to investigate on the effects of superoxide dismutase in 3T3 fibroblast and in experimental gingivitis in the rats. The effect of superoxide dismutase(SOD) to cell morphology and cell activity was measured in cultured mouse 3T3 fibroblast. After experimental gingivitis were induced by lipopolysaccharide(LPb) and bovine serum albumin(BSA), injection of SOD were done. WBC count and histologic findings were observed at 1, 2, 3, and 7 days. The results were as follows; 1. There was a little difference between LPS treated groups and SOD treated groups in 3T3 fibroblast morpholoy. 2. There was no difference between only SOD treated groups (except SOD 150U at 3days) and control in 3T3 fibroblast activity. 3. LPS $0.5{\mu}g/ml$ and SOD treated groups (except 150U) had decreased 3T3 fibroblast activity and no significant difference at 3 days. 4. LPS $5.0{\mu}g/ml$ and SOD treated groups were significantly increased cell activity of 3T3 fibroblast than control group at 1 day(P<0.05). 5. In LPS induced gingivitis, the number of leukocytes in SOD treated was significantly decreased than in saline treated at 1 day(P<0.05). 6. In histopathologic findings of LPS or BSA induced gingivitis, inflammatorycell infiltration in SOD treated groups were less than in saline treated group at 1, 2 and 3 days.

• 대한의생명과학회지
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• 제13권1호
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• pp.55-60
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• 2007

Computerized image processing and analysis system was used for quantitative assessment of skin tissue components in color histological sections. The purpose of this study was to determine that the changes in the collagen fiber density and elastic fiber density in dermis in the rat skin as aging. And also to determine that the correlation between fibroblast density and collagen fiber density and elastic fiber density in the aged rat skin. Ten weeks old ($130{\sim}150g$) eight and fifty-four weeks old ($300{\sim}350g$) eight female Sprague-Dawley rats were used. The full-thickness skin biopsy specimens were prepared serial sections and stained with hematoxylin and eosin, Masson's trichrome and Verhoeff-van Gieson. The collagen fiber and the elastic fiber were identified using the image analysis processing system and then calculated the collagen fiber density rate and the elastic fiber density rate in the dermis. It also identified fibroblast and calculated fibroblast density in the dermis. By using a Student's t-test, a decrease in the collagen fiber density rate (t=-4.650, P<0.001) and the elastic fiber density rate (t=-6.494, P<0.001) of dermis can be observed in aged rats as compared with the young rats. A Student's t-test showed a significantly less fibroblast density in the aged rats than the young rats (t=-4.497, P<0.001). There were significantly positive correlation between the fibroblast density and the collagen fiber density rate (r=.69, P<0.001) and the elastic fiber density rate (r=.91, P<0.001). These results indicate that the aging may decrease the collagen fiber density and elastic fiber density due to reduced the proliferative and synthetic activity of fibroblast in the dermis.

• 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.

• 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.

• 대한기계학회:학술대회논문집
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• 대한기계학회 2008년도 추계학술대회A
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• pp.1585-1587
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• 2008

Fibroblast is constantly subjected to mechanical loads in connective tissues where mechanical signals are converted to intercellular biochemical events. The aim of this study is to understand the effects of tensile stress on the neurotrophin (NT) and transforming growth factor (TGF) expression of fibroblast in vitro. Nerve growth factor (NGF) stimulates fibroblast migration, and TGF is related to tissue repair. In this study, at the uniaxial stretch of 10% strain and frequency of 0.5 Hz, different resting times of 0, 20, and 60 min are placed in between 10 min stimulations periods. Results show increase in NGF mRNA levels and a substantial decrease in NT3 mRNA after 1 hr of stimulation, indicating that the tensile stress may regulate NGF and NT3, key factors for the neurocosmetic applications. The mRNA level for TGF-${\alpha}$ and TGF-${\beta}2$ had increased up to two-folds after 1 hr of stimulation, showing that the tensile stress may control TGF, an important part of wound healing.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제33권3호
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• pp.211-220
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• 2007

Objective: In organotypic culture of immortalized human oral keratinocytes (IHOK), the change of the growth and differentiation was investigated according to the fibroblast type and the involvement of mitogen-activated protein (MAP) kinase. Materials & Methods: IHOK was cultured three dimensionally with gingival fibroblast (GF), dermal fibroblast (DF) and immortalized gingival fibroblast (IGF). We characterized biologic properties of three dimensionally reconstructed IHOK by histological, immunohistochemical, and Western blot analysis. We also investigated whether MAP kinase pathway was involved in epithelial-mesenchymal interaction by Western blot analysis. Results: The best condition of three dimensionally cultured IHOK was the dermal equivalent consisting of type I collagen and IGF. IGF increased the expression of more proliferating cell nuclear antigen (PCNA), involucrin than GF and DF in response to co-culture with IHOK. Extracellularly regulated kinase (ERK) pathway was activated in organotypic co-culture with IGF. Conclusion: The organotypic co-culture of IHOK with dermal equivalent consisting of type I collagen and IGF resulted in excellent morphologic and immunohistochemical characteristics and involved ERK pathway. The epithelial-mesenchymal interaction was activated according to the fibroblast type.