• Journal of Periodontal and Implant Science
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• v.47 no.2
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• pp.66-76
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• 2017

Purpose: Oral wound healing requires gingival fibroblasts to respond to local growth factors. Epigenetic silencing through DNA methylation can potentially decrease the responsiveness of gingival fibroblasts to local growth factors. In this study, our aim was to determine whether the inhibition of DNA methylation sensitized gingival fibroblasts to transforming growth factor-${\beta}1$ (TGF-${\beta}1$). Methods: Gingival fibroblasts were exposed to 5-aza-2'-deoxycytidine (5-aza), a clinically approved demethylating agent, before stimulation with TGF-${\beta}1$. Gene expression changes were evaluated using quantitative polymerase chain reaction (PCR) analysis. DNA methylation was detected by methylation-sensitive restriction enzymes and PCR amplification. Results: We found that 5-aza enhanced TGF-${\beta}1$-induced interleukin-11 (IL11) expression in gingival fibroblasts 2.37-fold (P=0.008). 5-aza had no significant effects on the expression of proteoglycan 4 (PRG4) and NADPH oxidase 4 (NOX4). Consistent with this, 5-aza caused demethylation of the IL11 gene commonly next to a guanosine (CpG) island in gingival fibroblasts. The TGF-${\beta}$ type I receptor kinase inhibitor SB431542 impeded the changes in IL11 expression, indicating that the effects of 5-aza require TGF-${\beta}$ signaling. 5-aza moderately increased the expression of TGF-${\beta}$ type II receptor (1.40-fold; P=0.009), possibly enhancing the responsiveness of fibroblasts to TGF-${\beta}1$. As part of the feedback response, 5-aza increased the expression of the DNA methyltransferases 1 (DNMT1) (P=0.005) and DNMT3B (P=0.002), which are enzymes responsible for gene methylation. Conclusions: These in vitro data suggest that the inhibition of DNA methylation by 5-aza supports TGF-${\beta}$-induced IL11 expression in gingival fibroblasts.

• Journal of Periodontal and Implant Science
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• v.32 no.2
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• pp.291-302
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• 2002

In most of the previous studies, invasive and discrete techniques have been used to monitor the healing process of the gingival graft. However, Laser Doppler Flowmetry(LDF, floLAB(R), Moor Instruments Ltd., England) is a non-invasive technique for measurement of blood flow in the tissue and also allows continuous monitoring. Thus, we tested the usefulness of LDF in monitoring the healing process of free gingival graft at gingival recession. Eleven gingival graft site of 7 patients, including 5 males and 2 females, aged between 21 and 41 years (mean age 28.5) were monitored for the blood flow. The blood flow in gingival graft at coronal site, central site, apical site, mesial site and distal site was measured using LDF. Blood flow was measured at 1- week, 2- week, 3- week and 4- week after gingival graft surgery from 10 a.m. to 2 p.m. Time-course of the healing process was evaluated by statistical analysis using repeated ANOVA and Duncan test. The results were as follows : (1) Blood flow stayed increased for 2 weeks, and then, it was a tendency to decrease. (2) The blood flow at distal site had always higher than mesial site during the measuring periods. (3) The blood flow was high orderly after 1 week ; most coronal site, most apical site, central site. But that was high orderly after 2 week, 3 week, 4 week ; most coronal site, central site, most apical site. In conclusion, LDF was a useful and clinically adaptable method to monitor wound healing process. Our study suggested that it was important to protect surgical site to promote initial wound healing.

• Journal of Periodontal and Implant Science
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• v.31 no.1
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• pp.135-148
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• 2001

Gingival fibroblasts are major cellular component of gingiva. However, the molecular mechanisms of senescence of human gingival fibroblasts are unknown. Human fibroblasts undergo replicative senescence in vitro after a limited number of population doublings. A reduced rate of proliferation is a prominent phenomenon observed in senescent fibroblasts. This phenomenon is controled by cell cycle regulatory proteins. The purpose of present study was to investigate the effect of replicative senescence on cell cycle progression and to find out its molecular mechanisms in human gingival fibroblasts. Replicative senescence of gingival fibroblasts were induced by subsequent cultures that were repeated up to 18 passage. In the present study, I examined change of cell proliferation, cell activity, cell viability and cell cycle progression during the replicative process. Also, I examined expression of cell cycle regulatory proteins which was estimated by western blot analysis. Cell proliferation, cell activity and cell viability of gingival fibroblasts were notably decreased with increase of population doubling level(PDL). S phase was decreased and G1 phase was increased with increase of PDL. Western blot analysis showed that levels of P16, p21 and p53 of senescent gingival fibroblasts(PDL41, PDL58) were higher than young fibroblasts(PDL27) and cdk4 were lower than young fibroblasts(PDL27). In conclusion, these results suggest that proliferative function of human gingival fibroblasts may be decreased by replicative senescence and its molecular mechanisms may be activatied with p16, p21, p53 and pRB, and repressed wtih cdk4.

• Journal of Periodontal and Implant Science
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• v.25 no.2
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• pp.181-191
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• 1995

The ability of fibroblasts attach to teeth is of paramount imporance in re-establishing the lost connective tissue attachment after periodontal therapy. Tobacco contains a complex mixture of substances including nicotine. various nitrousamines, trace elements. and a variety of poorly characterized substances. The effects of nicotine on fibroblasts have reported an altered morphology and attachment of fibroblasts to substrates and disturbances in protein synthesis and secretion. This study examined the effect of nicotine, a major component of the particulate phase of tobacco smoke, on human gingival fibroblasts and periodontal ligament cells attachment to tissue culture surfaces and cellular activity of human gingival fibroblasts and periodontal ligament cells. Pooled human gingival fibroblasts made from extraction of 3rd molar were utilized between passage 4 and 5 and plated in 96 well plate at 20,000 cells per well. Cell number were determined using 3-(4,5-dimethylthiazole-2-y)2,5-diphenyltetrazolium bromide(MTI) , which is reflection of mitochondrial dehydrogenase activity. The concentration of nicotine used were 0.025, 0.05, 0.1, 0.2 and $0.4{\mu}M$, the average serum concentration for a smoker being approximately $0.1{\mu}M$. The results were as follows : 1. Attachment effects of nicotine on human gingival fibroblasts and periodontal ligament cells Excepts of $0.4{\mu}M$, the effects on attachment with increasing numbers of cells attaching with increasing nicotine concentrations, compared to control group. But over the 60min, return to control value. 2. The effect of cellular activity on human gingival fibroblasts and periodontal ligament cells. The cellular activity of human gingival fibroblasts and periodontal ligament cells were similar or decrease to control value at 1st incubation day. At 2nd incubation day, 0.05, 0.1, 0.2, $0.4{\mu}M$ concentrations were statistically different from control value on gingival fibroblasts group. But at 3rd incubation day, cellular activities of all experimental group were significantly decrease than control group.

• Journal of Korean society of Dental Hygiene
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• v.14 no.6
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• pp.921-926
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• 2014

Objectives: The purpose of the study is to investigate the factors associated with gingival bleeding(GB) by tooth brushing in college students. Methods: A self-reported questionnaire was filled out by 232 college students in Daegu Health College from March to June, 2014. Data were analyzed for frequency, chi square test, and logistic regression analysis using SPSS 12.0 program. The study was a cross sectional study. The questionnaire consisted of general characteristics of the subjects(gender, age, marital status, and smoking), frequency and duration of tooth brushing, scaling experience, and physical health status. Self-reporting hemorrhage was reported by yes or no. Frequency of tooth brushing was documented as the number of behavior. Above 4 times of tooth brushing was defined as 4. Duration of tooth brushing was documented as minute. Above 4 minutes, it was recorded as 4. In physical health status, 1 is feeling weak and 4 is feeling very healthy. Cronbach alpha was 0.82 in the study. Results: There were significant relationships between gingival bleeding and age(p<0.05), subjective health(p<0.01), tooth brushing frequency(p<0.05) and duration(p<0.05) by chi square test. Logistic regression analysis showed that the age(p<0.05), subjective health(p<0.01), tooth brushing frequency(p<0.05) and duration(p<0.05) were associated with gingival bleeding. Prevalence of gingival bleeding in 20 years was 0.62(odds ratio 1.85, 95% CI 1.00~3.43) and it was higher than that in 10 years. Prevalence of gingival bleeding in good health group was -1.38 and it was lower than that in poor health group. Conclusions: The factors associated with gingival bleeding were age, subjective health, and tooth brushing frequency and time.

• Journal of Periodontal and Implant Science
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• v.32 no.1
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• pp.235-247
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• 2002

The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of human gingival fibroblasts. Primary culture of human gingival fibroblasts were grown in Dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells were inoculated in the multiwell plates coated with chitosan in concentration of 0.02, 0.2, and 2 mg/ml. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized nodules was evaluated after 21 days of culture. The results were as follows : The morphology of cells on the chitosan-coated well was round or spheric. Round cells were aggregated since 6 hours of culture and showed nodule-like appearance after 24 hours of culture and did not achieved confluency at 7 days. The attachment of gingival fibroblasts was inhibited by chitosan coating with a tendency of dose dependent pattern. But, cellular activity of unit cell was higher than control. The proliferation of gingival fibroblasts was inhibited by chitosan coating at 2 mg/ml(P<0.01), while the cell proliferation at 0.02, 0.2 $mg/m{\ell}$ was comparable to the control well. Total alkaline phosphatase activity was inhibited by chitosan coating and decreased in the course of time. While ALP activity of unit cell was the highest at 2mg/ml after 4 days of culture. Finally, gingival fibroblasts produced the mineralized nodule at 2 mg/ml. In summary, the attachment, proliferation, and alkaline phosphatase activity of gingival fibroblasts were influenced differently by the concentration of coated chitosan. From this study, it could be used as the matrix of tissue engineering for gingiva without inhibition on proliferation of gingival fibroblasts using chitosan at the optimal concentration (0.02mg/ml).

• Journal of Periodontal and Implant Science
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• v.35 no.4
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• pp.1109-1116
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• 2005

The purpose of this study was to observe the effects of periodontal therapy, including nonsurgical periodontal therapy with azithromycin, surgical therapy, and maintenace therapy on the drug-induced gingival enlargement, by means of measuring gingival thickness. The test group of 18 patients with drug-induced gingival enlargement received scaling, root planing with azithromycin for 5 days, with or without surgical periodontal treatment. The control group of 18 patients who had not taken any medication, received scaling and root planing, with or without surgical periodontal treatment. Both groups received supportive periodontal therapy every 3 months for 2 years. The mean period of total treatment is 32 months in the test group and 31 months in the control group. The thickness of the buccal gingiva was measured using an ultrasonic device of $SDM^{(R)}$(Krupp Corp., Essen, Germany). The results revealed that the test $group(1.21{\pm}0.51mm)$ showed statistically thicker buccal gingiva than the control $group(1.01{\pm}0.3mm)$. In the test group, the buccal gingiva was thickest on 2nd molars and was thinnest on canines of both dental arches. In the control group, the buccal gingiva was thickest on central incisors in the maxilla and 2nd molars in the mandible, while the thinnest areas were on canines in the maxilla and 1st premolars in the mandible. It would be concluded that the periodontal treatment with azithromycin aids in decreasing the degree of the gingival enlargement but cannot prevent the recurrence completely.

• Journal of Periodontal and Implant Science
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• v.36 no.4
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• pp.817-827
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• 2006

Purpose : This study was designed 1) to compare the perception of dental professionals and lay people with respect to minor variations in maxillary anterior tooth size and alignment and their relation to the surrounding soft tissues, and 2) to evaluate the normal tooth-gingiva topographical relationships in periodontally healthy young subjects, Materials and methods : Maxillary anterior teeth were intentionally diagrammed in varying degree of deviation with respect to one of three common anterior esthetic discrepancies including variations in crown length, shape of gingival margin, and length of interproximal contact, 17 images were generated to be preferentially selected by 2 groups consisting of dental professionals and lay people (total of 740). Smiling photographs of 120 dental students who had healthy periodontium were taken and the photographic images were analyzed to be classified as 17 kinds of altered image groups. Results : The results demonstrated noticeable difference between the varying levels of discrepancy. Both group preferred gingival margin of lateral incisor to be 0.5mm lower than that of central incisor. Lay people preferred the gingival margin shape that has 2/9 horizontal component of the crown width, while dental professionals preferred the gingival margin shape that has 1/9 horizontal component of the crown width. Lay people preferred longer length of the interproximal contact (two thirds of the crown length), whereas dental professionals preferred shorter length of the interproximal contact (half of the crown length). Photographic analysis of normal esthetic gingival topography revealed 2/9 horizontal component and short length of the interproximal contact which was of the hybrid nature of the preferences shared by lay people and dental professionals. Conclusion: The results of this study show that dental professionals and lay people demonstrated significant difference in their preference of dental esthetic components, which may then influence the decision making process by dental professionals with respect to designing the anterior esthetic gingival line.

• Journal of Periodontal and Implant Science
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• v.50 no.4
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• pp.226-237
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• 2020

Purpose: This study was conducted to assess the efficacy of prophylactic gingival grafting in the mandibular anterior labial area for preventing orthodontically induced gingival recession. Methods: Eight mongrel dogs received gingival graft surgery at the first (I1) and third (I3) mandibular incisors on both sides based on the following group allocation: AT group (autogenous connective tissue graft on I1), AT-control group (contralateral side in the AT group), CM group (xenogeneic cross-linked collagen matrix graft on I3) and CM-control group (contralateral side in the CM group). At 4 weeks after surgery, 6 incisors were splinted and proclined for 4 weeks, followed by 16 weeks of retention. At 24 weeks after surgery, casts were made and compared with those made before surgery, and radiographic and histomorphometric analyses were performed. Results: Despite the proclination of the incisal tip (by approximately 3 mm), labial gingival recession did not occur. The labial gingiva was thicker in the AT group (1.85±0.50 mm vs. 1.76±0.45 mm, P>0.05) and CM group (1.90±0.33 mm vs. 1.79±0.20 mm, P>0.05) than in their respective control groups. Conclusions: The level of the labial gingival margin did not change following labial proclination of incisors in dogs. Both the AT and CM groups showed enhanced gingival thickness.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.28 no.5
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• pp.445-453
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• 2006

Cyclosporine A (CsA) is a powerful immunosuppresive agent used to prevent graft rejection of organ and treat autoimmune disease. One of the major side effects associated with CsA treatment is the development of gingival overgrowth. The purpose of this study was to investigate the mRNA expression and association of the several growth factors in gingival overgrowth induced by CsA, respectively. Gingival fibroblasts were obtained from gingival tissues of healthy donor and the patients treated with CsA. The cultured gingival fibroblasts were incubated with increasing concentrations of CsA for 24 hours, and the expression of MMP-1, TIMP-1, $TGF-{\beta}_1$, p21 were determined by reverse transcription-polymerase chain reaction (RT-PCR). The expressions of MMP-1 was slightly increased according to the concentration of treated CsA, but there was no statistical significance. TIMP-1 showed the increased expression at the CsA concentration of 250 and 500 ng/ml and significantly decreased at the CsA concentration of 750ng/ml. $TGF-{\beta}_1$ showed the increased expression at the CsA concentration of 500 and 750 ng/ml. The expression of p21 was not changed significantly. We concluded that the gingival hyperplasia induced by CsA was more related with $TGF-{\beta}_1$ than MMP-1 or TIMP-1 on gingival collagen metabolism in patients treated with CsA.