• Journal of Periodontal and Implant Science
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• 제32권1호
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• pp.129-142
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• 2002

Epithelial-mesenchymal interaction plays a important role in cell growth and differentiation. This interaction is already well known to have an importance during the organ development as well as cell growth and differentiation. However, in vitro experimental model is not well developed to reproduce in vivo cellular microenvironment which provide a epithelial-mesenchymal interaction. Because conventional monolayer culture lacks epithelial-mensenchymal interaction, cultivated cells have an morphologic, biochemical, and functional characteristics differ from in vivo tissue. Moreover, it's condition is not able to induce cellular differention due to submerged culture condition. Therefore, the aims of this study were to develop and evaualte the in vitro experimental model that maintains epithelial-mesenchymal interaction by organotypic raft culture, and to characterize biologic properties of three-dimensionally reconstituted oral keratinocytes by histological and immunohistochemical analysis. The results were as follow; 1. Gingival keratinocytes reconstituted by three-dimensional organotypic culture revealed similar morphologic characteristics to biopsied patient specimen showing stratification, hyperkeratinosis, matutation of epithelial architecture. 2. Connective tissue structure was matured, and there is no difference during stratification period of epithelial 3-dimensional culture. 3. The longer of air-exposure culture on three-dimensionally reconstituted cells, the more epithelial maturation, increased epithelial thickness and surface keratinization 4. In reconstitued mucosa, the whole epidermis was positively stained by anti-involucrin antibody, and there is no difference according to air-exposured culture period. 5. The Hsp was expressed in the epithelial layer of three-dimensionally cultured cells, especially basal layer of epidermis. The change of Hsp expression was not significant by culture stratification. 6. Connexin 43, marker of cell-cell communication was revealed mild immunodeposition in reconstitued epithelium, and there is no significant expression change during stratification. These results suggest that three-dimensional oragnotypic co-culture of normal gingival keratinocytes with dermal equivalent consisting type I collagen and gingival fibroblasts results in similar morphologic and immunohistochemical characteristics to in vivo patient specimens. And this culture system seems to provide adequate micro-environment for in vitro tissue reconstitution. Therefore, further study will be focused to study of in vitro gingivitis model, development of novel perioodntal disease therapeutics and epithelial-mensenchymal interaction.

• Journal of Periodontal and Implant Science
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• 제43권1호
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• pp.3-11
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• 2013

Periodontitis is a chronic inflammation of periodontal tissue caused by subgingival plaque-associated bacteria. Periodontitis has long been understood to be the result of an excessive host response to plaque bacteria. In addition, periodontal pathogens have been regarded as the causative agents that induce a hyperinflammatory response from the host. In this brief review, host-microbe interaction of nonperiodontopathic versus periodontopathic bacteria with innate immune components encountered in the gingival sulcus will be described. In particular, we will describe the susceptibility of these microbes to antimicrobial peptides (AMPs) and phagocytosis by neutrophils, the induction of tissue-destructive mediators from neutrophils, the induction of AMPs and interleukin (IL)-8 from gingival epithelial cells, and the pattern recognition receptors that mediate the regulation of AMPs and IL-8 in gingival epithelial cells. This review indicates that true periodontal pathogens are poor activators/suppressors of a host immune response, and they evade host defense mechanisms.

• Journal of Periodontal and Implant Science
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• 제27권2호
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• pp.291-303
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• 1997

Periodontal ligament cells may have a role in the regulation of hard and soft periodontal tissues, but their specific function has not yet to be determined. To evaluate further their role in periodontal regeneration, they were examined for osteoblast-like behavior. Periodontal ligament cells and gingival fibroblasts were primarily cultured from extracted premolar with non-periodontal diseases. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidity incubator, and as a measure of cell characterization, it was examined that the morphology, alkaline phosphatase activity, collagen synthesis, and immunocytochemistry for osteonectin, osteocalcin, and collagen type I. Healthy periodontal ligament cells has more osteoblastic-like cell property in alkaline phosphatase activity. and collagen synthesis than gingival fibroblast. Immunocytochemistry localization explained that calcitonin were expressed in periodontal ligament cells only, and osteonectin and type I collagen were produced in both cells simultaneously. This results indicate that the growth characteristics of periodontal ligament cells and gingival fibroblasts exhibit some differences in proliferative rates and biochemical synthesis. The differences may help to calrify the role such cells play in the regenearation of periodontal tissues.

• Journal of Periodontal and Implant Science
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• 제25권3호
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• pp.741-755
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• 1995

구강내 매식된 임프란트가 과도한 교합력이나 염증등의 이유로 구강내로 노출되었을 때 세균독소에 이완된 면을 제거하고 평활한 면을 형성하여 주위의 연조직, 경조직에 적합한 상태로 만들어 건강한 상태로 구강내에 유지하기 위해서 임프란트 매식체 표면을 기계적인 표면처리방법으로 처치하여 이러한 방법이 임프란트 표면성분, 치은섬유아세포의 전개양상에 미치는 영향을 알아보고자 본 실험을 실행하였다. IMZ사에서 제작한 직경 10mm, 높이 2mm의 원판 타이타늄을 이용하여 피막되지 않은 타이타늄면과 TPS면을 대조군으로 하고 기계적인 표면처리방법인 low speed stone bur처치면을 실험군으로 설정한 후 EDX로 타이타늄 표면성분을 분석하였고 주사전자현미경으로 치은섬유아세포의 전개양상을 관찰하였다. EDX에 의한 타이타늄 표면성분분석 결과 모든 실험군에서 titanium peak, 소량의 aluminum이 나타났으며 그외의 성분은 나타나지 않았다. 치은섬유아세포의 전개양상에 대한 주사전자현미경 관찰결과 평활한 타이타늄면에서 접종 30분 후 세사상돌기와 박판엽상으로 확장된 세포가 많이 관찰되며 6시간 후 신장된 치은섬유아세포가 시편에 밀착된 양상을 보였고 24시간 후 치은섬유아세포는 시편의 모든 면을 피개하며 가공시의 평행한 선을 따라 방향성을 띄었다. TPS가 잔존한 stone처치군에서 세포 접종 30분 후 세사상돌기가 적게 관찰되어 평활한 타이타늄면에 비해 초기부착이 늦은 것을 알 수 있었고 6, 24시간후 치은섬유아세포는 거친면으로 인해 시편에 밀착되지 못한 양상을 보였으나 평활한 타이타늄면과 연결되며 시편의 모든면을 피개하였다. TPS군에서 치은섬유아세포는 세포 접종 30분후 세사상돌기를 거의 찾아 볼 수 없어 초기부착이 다른군에 비해 늦으며 세포배양 6, 24시간후에도 시편에 밀착되지 못하고 박판상돌기가 가늘고 길게 돌출되어 여러면에 부착된 양상을 보였으며 세포가 부착되지 않은 TPS면이 관찰되었다.

• The Journal of Advanced Prosthodontics
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• 제9권4호
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• pp.315-320
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• 2017

PURPOSE. The aim of this investigation was to analyze the dimensions of clinical crowns and to classify the crown and the gingival type in the anterior teeth in Korean young adults. MATERIALS AND METHODS. Casts were obtained from 50 subjects ranging in age from 24 to 32. Measurements of length and width were made on the casts using a pair of digital calipers on the entire dentition. Crown thickness and papilla height were also measured and MDW/CL (mesiodistal width to clinical length) and CW/CL (cervical width to clinical length) ratios of the maxillary anterior teeth were calculated. The K-clustering method was used for CW/CL to classify the anterior tooth shape into three groups (tapered, ovoid, and square), and one-way analysis of variance and Duncan's post-hoc comparison were used to evaluate statistical significance between the groups. Pearson's correlation analysis was performed between tooth shape and papillary height (PH) to demonstrate the correlation between tooth shape and gingival morphological characteristics. RESULTS. The average length of the maxillary central incisors was 9.89 mm; the mesio-distal width was 8.54 mm; and the ratio of width/length was 0.86 in Korean young adults. The average bucco-palatal thickness of the central incisor was 3.14 mm at the incisal 1/3 aspect. Ovoid type was the most common tooth shape (48%), followed by square type (29%) and taper type (23%) in the central incisors of Korean young adults. Tooth shape and gingival type were correlated with each other. CONCLUSION. New reference data were established for tooth size in Korean young adults and the data show several patterns of tooth shape and gingival type. Clinicians should diagnose and treat based on these characteristics for better results in the Korean population.

• Journal of Periodontal and Implant Science
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• 제37권4호
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• pp.849-857
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• 2007

Purpose: The purposes of this study were to compare and quantify the expressions of RANK and RANKL in the gingival tissues of non-periodontitis patient and patients with chronic periodontitis, in order to understand the contribution of these proteins to periodontal destruction. Material and methods: Gingival tissue samples were obtained during periodontal surgery or tooth extraction. According to the patient's systemic condition & clinical criteria of gingiva, each gingival sample was divided into two groups. Group 1 (n=8) is clinically healthy gingiva without bleeding and no evidence of bone resorption or periodontal pockets, obtained from non-periodontitis patients. Group 2 (n=8) is inflammed gingiva from patients with chronic periodontitis. Tissue samples were prepared and analyzed by Western blotting. The quantification of RANK and RANKL were performed using a densitometer and statistically analyzed by Student's t-Test. Results: The expression of RANK were similar in group 1 and 2. The difference between group 1 and 2 was not statistically significant. And the mean amount of RANKL was more increased in group 2 than group 1. The difference between group 1 and group 2 was statistically significant. Conclusion: The expression level RANK didn't show any significant difference between healthy tissue from non-periodontitis patients and inflamed tissue from chronic periodontitis, but the expression level of RANKL in inflammed tissue from chronic periodontitis showed significantly increased tendency compared to healthy gingiva from non-periodontitis patients. Therefore, characteristics of RANK and RANKL in progress of chronic periodontitis would be basis of further studies in diagnostic method and treatment index of the disease.

• 치위생과학회지
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• 제12권6호
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• pp.689-695
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• 2012

The aim of this study was to elucidate the effects of concentrated growth factors (CGFs) on human gingival fibroblasts in vitro. Blood was collected from three male volunteers (average age 27 years). CGFs were prepared using standard protocols. The CGF exudates were collected at the following culture time points: 1, 7, 14, and 21 days. The levels of platelet-derived growth factor BB (PDGF-BB) and transforming growth factor ${\beta}1$ (TGF-${\beta}1$) in CGFs were quantified. The CGF exudates were then used to culture human gingival fibroblasts. The biologic characteristics of these fibroblasts were analyzed in vitro for 21 days. Platelet-rich plasma released the highest amounts of TGF-${\beta}1$ and PDGF-BB on the first day. The level of TGF-${\beta}1$ had decreased slightly by day 7, although the difference compared to levels at day 1 was not statistically significant. However, by days 14 and 21, levels of TGF-${\beta}1$ had dropped significantly compared to day 1 levels. The levels of PDGF-BB at days 7, 14, and 21 did not differ significantly from that measured on day 1. CGFs maintained the release of autologous growth factors for a reasonable period of time (7 days for TGF-${\beta}1$ and 21 days for PDGF-BB). Gingival fibroblasts treated with CGF exudates collected at day 14 reached peak viability and synthesized type I collagen. Furthermore, the CGF exudates exerted positive effects on the proliferation and differentiation of these cells at days 1, 7, 14, and 21. The findings of this study suggest that treatment with CGFs represents a promising method of enhancing mucosal healing following surgical procedures.

• Journal of Periodontal and Implant Science
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• 제34권2호
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• pp.317-332
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• 2004

Osteoblasts from alveolar bone may have an important role in the bone regeneration for periodontium, but their culture and characterization are not determined yet. The purpose of this study was to investigate the biological characteristics of primary explant cultured osteoblasts(PECO) from alveolar bone. Osteoblasts were isolated and cultured from alveolar socket of extracted tooth in children. To compare the characteristics, osteoblasts and gingival fibroblasts were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, l00% humidity incubator, and human fetal osteoblasts cell line(hFOB1) were cultured with DMEM at $34^{\circ}C$, 5%, $CO_2$ 100% humidity incubator. To characterize the isolated bone cells, morphologic change, cell proliferation and differentiation were measured. Morphology of PECO was small round body or cuboidal shape on inverted microscope and was similar with hFOB1. PECO became polygonal shape with stellate and had an amorphous shape at 9th passage in culture. PECO had significantly higher activity than that of gingival fibroblasts and hFOB1 in alkaline phosphatase activity. The expression of osteocalcin and bone sialoprotein in PECO was notably increased when compared with hFOB1 and gingival fibroblasts. These result indicated that PECO from alveolar bone in children has an obvious characteristics of osteoblast, maybe applied for the regeneration of bone.

• Journal of Periodontal and Implant Science
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• 제26권1호
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• pp.143-160
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• 1996

The purpose of this study was to evaluate the response in aspect of attachment and growth rate of osteoblasts and growth rate of osteoblasts and human gingival fibroblasts to the commercially pure titanium(CP titanium)and titanium alloy(Ti-6AI-4V) that are used widely as implant materials, and to obtain the basic information to ideal implant materials. In the studly, commercially pure titanium in first test group, titanium alloy(Ti-6AI-4V) in second test group, cobalt-chrome-molybdenum alloy(Co-Cr-Mo alloy) in positive control group, and tissue culture polystyrene plate in negative control group were used. The results of this study were as follows. 1. Bone marrow cells cultured on CP titanium and Ti-6Al-4V showed significantly greater attachment and growth rate(p(0.05) compared to Co-Cr-Mo alloy in each time. 2. There were no significant differences(p>0.05) in attachment and growth rate of bone marrow cells cultured on CP titanium and Ti-6AI-4V or tissue culture plate. 3. Most bone marrow cells cultured on CP titanium, Ti-6Al-4V and tissue culture plate were attached well to each substratum in first 2days, and then, grew at higher growth rate. On the other hand, some cells cultured on Co-Cr-Mo alloy failed to attach in first 2 days, and then, attached cells grew at lower growth rate than other groups. 4. Attachment and growth rates of gingival fibroblasts cultured on CP titanium and Ti-6Al-4V showed no significant differences(p>0.05) compared to Co-Cr-Mo alloy in 2 days, but significantly greater increase(p<0.05) in 5 and 9 days. 5. There were no significantly differences(p>0.05) between growth rates on gingival fibroblasts cultured on CP titanium, Ti-6Al-4V and tissue culture plate in 2 and 5days, but a significant lower growth rate(p<0.05) on CP titanium and Ti-6Al-4V versus tissue culture plate. 6. Some gingival fibroblasts cultured on all specimen groups failed to attach, but attached cells grew well, especially on CP titanium, Ti-GAl-4V and tissue culture plate. 7. There were no significant differences(P>0.05) between growth rates of both bone marrow cells and gingival fibroblasts cultured on CP titanium and Ti-6AI-4V. As a result of this study, both commercially pure titanium and Ti-6AI-4V showed excellent biocompatibility and there was no significant difference in the cellular response to the both metals. Bone marrow cells cultured on each substratum showed significantly greater growth rate and responded sensitively to cytotoxic effects of metal surfaces compared to gingival fibroblasts. Considering cell response to the substrate, it was likely that the composition itself of titanium metals have no significant effect on the biocompatibility. Further study need to be done to evaluate the influence of surface characteristics on cellular responses.

• Journal of Periodontal and Implant Science
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• 제28권1호
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• pp.17-35
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• 1998

Chitosan, with a chemical structure similar to hyaluronic acid, has been implicated as a wound healing agent. The purpose of this research was to evaluate the effects of chitosan on the characteristics of periodontal ligament cells, calvaria cells and gingival fibroblasts and to define the effects of chitosan on bone formation in vitro. In control group, the cells were cultured alone with Dulbecco's Modified Eagle's Medium contained with 10% Fetal bovine serum, 100unit/ml penicillin, $100{\mu}g/ml$ streptomycin, $0.5{\mu}g/ml$ amphotericin-B. In experimental group, chitosan($40{\mu}g/ml$) is added into the above culture condition. And then each group was characterized by examining the cell proliferation at 1,3,5,7,9,12,15 day, the amount of total protein synthesis, alkaline phosphatase activity at 3, 7 day and the ability to produce mineralized nodules of rat calvaria cell at 11 day. The results were as follows : 1. At early time both periodontal ligament cells and calvaria cells in chitosan-treated group proliferated more rapidly than in non-treated control group, but chitosan-treated group of periodontal ligament cells at 9 days and calvaria cells at 12days showed lower growth rate than control group. Gingival fibroblast in chitosan-treated group had lower growth rate than in control group but the difference was not statistically significant (P< 0.01).2. Both periodontal ligament cells and calvaria cells in chitosan-treated group showed much protein synthesis than in control group at 3 days, but showed fewer than in control group at 7 days. Amount of total protein synthesis of gingival fibroblast didn't have statistically significant difference among the two groups(P< 0.01). 3. At 3 and 7 days, alkaline phosphatase activity of periodontal ligament cells and calvaria cells was increased in chitosan-treated group, but at 7 days there was not statistically significant difference among the two groups of calvaria cells (P< 0.01). Alkaline phosphatase activity of gingival fibroblast didn't have statistically significant difference among the two groups(P<0.01). 4. Mineralized nodules in chitosan-treated group of rat calvaria cells were more than in control group. In summery, chitosan had an effect on the proliferation, protein systhesis, alkaline phosphatase activity of periodontal ligament cells and calvaria cells, and facilitated the formation of bone. It is thought that these effects can be used clinically in periodontal regeneration therapy.