Gingival crevicular fluid (GCF) is a promising source for markers of destructive periodontal disease activity. This study was undertaken to evaluate the protein composition of GCF in varying stages of the gingival inflammatory response. GCF sampled from 26 people with clinically healthy gingiva and 18 people with periodontitis were examined via sodium dodecyl sulphate polyacrylamide gel electrophoresis(SDS/PAGE). The result were as follows. 1. Total amount of GCF protein of diseased group significantly different from that of normal group. But difference in protein concentration was not that significant. 2. In analyzing GCF with SDS/PAGE, it was suggested that albumin is used as indicator plasma protein leakage because of heavily staining bond of albumin in patients with periodontal disease. 3. In diseased group, overall bonds of protein and bands of high molecular weight protein were heavily stained. It was proved useful information on high molecular plasma protein leakage with increasing vascular permeability due to inflammation.
Root surface exposure due to gingival recession after periodontal surgery, dentin exposure after root planing elicit pain response when exposed to mechanical, heat, chemical or osmotic stimulation. Especially, patients treated with periodontal surgery, show high frequency and there have been reports showing the 1 out of 7 patients have dentin hypersensitivity. There have been many studies on the clinical effects of various materials on the treatment of dentin hypersensitivity. but, none could provide absolute clinical efficacy. In this study, 45 teeth from 30 patients, who had had periodontal surgery and showed dentin hypersensitivity after surgery were chosen for the experimental group and they were illuminated with laser, 15teeth were chosen for the control group and they were not exposed to laser. After this dentin hypersensitivity was elicited by tactile, compressed air, cold water and then, the degree was evaluated using NRS(Numerical Rating Scale). And during LLLT(Low Level Laser Therapy) semiconductor laser using Gallium - Arsenide as a diode was illuminated for 180 seconds at a frequency of 7(500Hz). This therapy was done 10 times, and each time the changes in dentin hypersensitivity was evaluated using NRS. The results were as follows : 1. After treat with LLLT on dentin hypersensitivity due to periodontal surgery, 22.2% showed total loss of dentin hypersensitivity, 60.0% showed loss of tactile dentin hypersensitivity, 48.8% showed loss of compressed air dentin hypersensitivity, 22.2% showed loss of cold water dentin hypersensitivity. 2. As a result of clinical evaluation of dentin hypersensitivity using NRS, there was significant increase in improvement of dentin hypersensitivity in the experimental group compare to the control group(P<0.05). And there was almost no natural loss of dentin hypersensitivity in the control group. 3. In comparison of the stages of evaluation, there was significant difference in between experimental and control group. after the second visit(P<0.05), and the difference increased with each visit.
The purpose of this study was to evaluate the effect of fibrin tissue adhesive and porous resorbable calcium carbonate on the periodontal regeneration of the class II furcation defect in dogs. Class II furcation defect was surgically created on the second, third, and fourth premolars bilaterally in the mandibles of six mongrel dogs. The experimental sites were divided into four groups according to the treatment modalities: Control-surgical debridement only; Group I-calcium carbonate grafting; Group II-application of fibrin adhesive only; Group III-application of fibrin adhesive after calcium carbonate grafting. The animals were sacrificed at the 2, 4, and 12 weeks after periodontal surgery and the decalcified specimens were prepared for histological and histometrical examination. The results are as follows : Clinically, there were no inflammatory response in all groups after 2, 4, 12 weeks. In the Control group, junctional epithelium was grown downward to the reference notch. In Group I, graft materials were exfoliated from the defect throughout the experimenta periods andnew bone was seen in the notch area at 4 and 12 week specimens. In Group II, fibrin adhesive was absorbed at 2 week specimens, and connective tissue attachment increased than that of control group. New cementum and new bone were seen above the notch area. In Group III, the graft material was maintained in the defect throughout the experimental period and inducing the amount of periodontal tissue regeneration was higher than other groups. These results suggest that the use of fibrin tissue adhesive in conjunction with porous resorbable calcium carbonate would improves the stability of graft material and inhibit the epithelial down growth and make it be a feasible method for periodontal regeneration.
Recent studies have demonstrated that human periodontal ligament cells express receptor activation of nuclear factor ${\kappa}B$ ligand (RANKL) which enhances the bone resorbing activity of osteoclasts differentiated from hematopoietic preosteoclasts. The purpose of this study is to determine the effects of p38 MAPK and JNK kinase upon regulating RANKL and OPG in response to $IL-1{\beta}$(l ng/ml) in rat periodontal ligament cells. Soluble RANKL was measured by immunoassay. The effects of p38 MAPK on RANKL and OPG expression was determined by RT-PCR. The results were as follows: 1. Periodontal ligament cells which stimulated by $IL-1{\beta}$ increased soluble RANKL synthesis by dose-dependent pattern. 2. p38 MAP kinase inhibitor (SB203580) showed regulation of soluble RANKL expression by dose-dependent manners. 3. p38 MAP kinase inhibitor (SB203580) regulated the expression of RANKL, but it dose regulate the expresseion of OPG. 4. JNK (c-jun $NH_2-terminal$ kinase) inhibitor (PD98059) did not regulate mRANKL and mOPG. These results suggested that p38 MAPK play a significant role in RANKL gene expression.
치주질환은 상당수의 성인들이 가지고 있는 질환이지만 아직 분자적인 수준에서의 발생 기작과 치료 방법에 대해서는 많은 것이 밝혀져 있지 않다. 본 연구에서는 치주질환 조직과 정상 조직에서 얻어진 유전자 발현 데이터를 이용하여 치주질환 조직과 정상 조직 사이에 분자적 차이가 있는지를 확인한다. 특히 기계학습 알고리즘을 이용하여 유전자 발현양 기반 치주질환 조직과 정상 조직의 분류가 가능한지를 확인하고, 각 조직에서 발현양 차이가 나는 유전자들이 주로 어떤 기능을 하는 것인지 살펴본다. t-SNE를 이용한 분석 결과 정상 조직과 치주질환 조직 샘플이 명확히 구분되어 군집화 될 수 있음이 확인되었다. 또한, 결정 트리, 랜덤 포레스트, 서포트 벡터 머신을 이용한 분류 알고리즘을 적용한 결과 불균형 데이터임에도 높은 정확도와 민감도, 특이도를 보였으며, 염증 반응 및 면역 반응 관련 유전자들이 주로 두 집단 간에 차이를 보임이 확인되었다.
Ahu Dikilitas;Fatih Karaaslan;Sehrazat Evirgen;Abdullah Seckin Ertugrul
Journal of Periodontal and Implant Science
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제52권6호
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pp.455-465
/
2022
Purpose: Periodontal diseases are inflammatory conditions that alter the host's response to microbial pathogens. Type 2 diabetes mellitus (T2DM) is a complex disease that affects the incidence and severity of periodontal diseases. This study investigated the gingival crevicular fluid (GCF) levels of colony-stimulating factor-1 (CSF-1) and interleukin-34 (IL-34) in patients with stage III grade C periodontitis (SIII-GC-P) and stage III grade C periodontitis with uncontrolled type 2 diabetes (SIII-GC-PD). Methods: In total, 72 individuals, including 24 periodontally healthy (PH), 24 SIII-GC-P, and 24 SIII-GC-PD patients, were recruited for this study. Periodontitis patients (stage III) had interdental attachment loss (AL) of 5 mm or more, probing depth (PD) of 6 mm or more, radiographic bone loss advancing to the middle or apical part of the root, and tooth loss (<5) due to periodontal disease. Radiographic bone loss in the teeth was also evaluated; grade C periodontitis was defined as a ratio of the percentage of root bone loss to age greater than 1.0. The plaque index (PI), gingival index (GI), presence of bleeding on probing (BOP), PD, and clinical AL were used for clinical periodontal assessments. GCF samples were obtained and analyzed using an enzyme-linked immunosorbent assay. Results: All clinical parameters-PD, AL, GI, BOP, and PI-were significantly higher in the SIII-GC-PD group than in the PH and SIII-GC-P groups for both the full mouth and each sampling site (P<0.05). The total IL-34 and CSF-1 levels were significantly higher in the SIII-GC-PD group than in the PH and SIII-GC-P groups (P<0.05), and there were significant differences between the periodontitis groups (P<0.05). Conclusions: These findings suggest that IL-34 and CSF-1 expression increases in patients with SIII-GC-PD. CSF-1 was associated with the inflammatory status of periodontal tissues and T2DM, while IL-34 was associated only with T2DM.
Purpose: The aim of this study was to investigate the efficacy of photo-crosslinked gelatin methacryloyl (GelMa) hydrogel containing calcium phosphate nanoparticles (CNp) when applying different fabrication methods for bone regeneration. Methods: Four circular defects were created in the calvaria of 10 rabbits. Each defect was randomly allocated to the following study groups: 1) the sham control group, 2) the GelMa group (defect filled with crosslinked GelMa hydrogel), 3) the CNp-GelMa group (GelMa hydrogel crosslinked with nanoparticles), and 4) the CNp+GelMa group (crosslinked GelMa loaded with nanoparticles). At 2, 4, and 8 weeks, samples were harvested, and histological and micro-computed tomography analyses were performed. Results: Histomorphometric analysis showed that the CNp-GelMa and CNp+GelMa groups at 2 weeks had significantly greater total augmented areas than the control group (P<0.05). The greatest new bone area was observed in the CNp-GelMa group, but without statistical significance (P>0.05). Crosslinked GelMa hydrogel with nanoparticles exhibited good biocompatibility with a minimal inflammatory reaction. Conclusions: There was no difference in the efficacy of bone regeneration according to the synthesized method of photo-crosslinked GelMa hydrogel with nanoparticles. However, these materials could remain within a bone defect up to 2 weeks and showed good biocompatibility with little inflammatory response. Further improvement in mechanical properties and resistance to enzymatic degradation would be needed for the clinical application.
In infectious disease, invasion of host tissue by bacteria or their products frequently induces a wide variety of inflammatory and immunopathologic reaction. Evidence indicates that cytokines are involved in the initiation and progression of chronic inflammatory diseases, such as periodontitis. Interleukin-6, which is a multifunctional cytokine, has important roles in acute and chronic inflammation and may also be implicated in bone resorption. Periodontal diseases are characterized by chronic inflammation of the periodontium with alveolar bone resoption. A principal driving force behind this response appears to lie in the immune system's response to bacteria. Many of the cell components which have been shown to function as virulence factors in gram-negative bacteria are associated with the bacterial surface. Of these, lipopolysaccharide has been characterized as one that mediates a number of biological activities which can lead to the destruction of host tissue. Non-steroidal antiinflammatory drug is used for reduce inflammation, and most of NSAIDs inhibit prostaglandine $E_2$ production, but it is shown that $PGE_2$ production is stimulated by IL-1 in recent study. So, the influence of other cytokines except $PGE_2$ on periodontium can not be avoided. Therefore, new antiinflammatory drug is needed. Rhizoma coptidis is used in oriental medicine for anti-inflammation and antiseptics. In this present study, we examined the IL-6 release in periodontal ligament cells treated with the lipopolysaccharide, and also the effect of rhizoma coptidis on cellular activity and IL-6 production of periodontal ligament cells. To evaluate the effect of rhizoma coptidis on cellular activity, the cells were seeded at a cell density of $1{\times}10^4$ cells/well in 24-well culture plates. After one day incubation, 1-6, 10-9 and 10-12 g/ml of rhizoma coptidis and 5, $10{\mu}g/ml$ of LPS were added to the each well and incubated for 1 and 2 days, respectively. Then, MTT assay were carried out. To evaluate the effect of rhizoma coptidis on IL-6 production, the cells were seeded at a cell density of $1.5{\times}10^4$ cells/well in 24-well culture plates. After one day incubation, 10-9 g/ml of rhizoma coptidis and 5, $10{\mu}g/ml$ of LPS were added to the each well and incubated for 3, 6, 12 and 24 hours. Then, amounts of IL-6 production is measured by IL-6 ELISA kit used. The results were as follows : 1. Rhizoma coptidisrbelow to ($10^{-6}g/ml$) significantly increaed cellular activity of periodontal ligament cells than control. 2. Rhizoma coptidist ($10^{-9}g/ml$) significantly increased cellular activity of LPS($5{\mu}g/ml$)-treated periodontal ligament cells than control. 3. LPS(5 and $10{\mu}g/ml$) significantly increased IL-6 production of periodontal ligament cells than control. 4. Rhizoma coptidis($10^{-9}g/ml$) decreased IL-6 production of LPS ($5{\mu}g/ml$)-treated periodontal.ligarnent cells than LPS only tested group. These findings suggest that stimulation of the IL-6 release of periodontal ligament cells by LPS may have a role in the progression of inflammation and alveolar bone resoption in periodontal disease, and that inhibition of the IL-6 release of cells and stimulation of cellular activity by rhizoma coptidis may help the periodontal regeneration.
Diabetes mellitus is a systemic disease with profound effects on oral health and periodontal wound healing. Uncontrolled diabetes adversely affects surgical wound healing and is often associated with abnormal proliferation of fibroblasts. Human gingival fibroblasts and PDL cells were chosen because they are intimately involved in periodontal therapy and are important for the success of surgical procedure such as guided tissue regeneration. The aim of the present study was to elucidate whether cellular activity and collagen synthesis by glucose pre-treated human gingival fibroblasts and PDL cells are influenced by insulin, and whether healthy cells differ from glucose treated cells. Cells were cultured with DMEM at $37^{\circ}C$, 5% $CO_2$, 100% humidified incubator. To evaluate the effect of glucose on gingival fibroblasts and periodontal ligament cells, the cells were seeded at a cell density of $1{\times}10^4\;cells/well$ culture plates and treated with 20 and 50mM of glucose for 5 days. Then MTT assay was carried out. To evaluate the effect of insulin on glucose-pretreated cells, the cells were seeded at a cell density of $1{\times}10^4\;cells/well$ culture plates and treated with 20 and 50mM of glucose for 5 days. After incubation, $10^3$, $10^4$ and $10^5mU/l$ of insulin were also added to the each well and incubated for 2 days, respectively. Then, MTT assay and collagen synthesis assay were carried out. The results indicate that cellular activity of gingival fibroblasts significantly increased by glucose while periodontal ligament cells were unaffected and cellular activity of gingival fibroblasts and periodontal ligament cells were unaffected by insulin. Collagen synthesis of gingival fibroblast with 20mM glucose and insulin unaffected, but 50mM glucose and insulin increased than control. Collagen synthesis of periodontal ligament cell with 20mM glucose and $10^5mU/l$ insulin significantly increased than other groups and 50mM glucose pretreated PDL cells significantly increased at $10^3mU/l$ insulin but decreased at $10^4mU/l$ insulin. Our findings indicated that these cell types differed in their growth response to glucose, and the increase in collagen synthesis was significantly raised at insulin level of $10^3mU/l$ in gingival fibroblasts and periodontal ligament cells except 20mM glucose pretreated periodontal ligament cells.
Introduction : The purpose of this study was to evaluate the possibility of the acellular dermal matrix (ADM) as a barrier membrane for bone regeneration, and to evaluate the osteogenic effect of ADM as a carrier system for rhBMP-2 in the rat calvarial defect model. Materials and Methods: An 8-mm, calvarial, critical-size osteotomy defect was created in each of 60 male Spraque-Dawley rats(weight $250{\sim}300g$). Three groups of 20 animals, each received either rhBMP-2(0.025mg/ml) in an ADM carrier, ADM only, or negative surgical control. And each group was divided into 2- and 8-weeks healing intervals. The groups were evaluated by histologic and histomorphometric parameters(10 animals/group/healing intervals). Data were expressed as $means{\pm}standard$ deviations($m{\pm}SD$). Comparisons between experimental and control groups were made using two-way ANOVA and post hoc t-test. Comparisons between 2 weeks and 8 weeks were made using paired t-test. The level of statistical difference was defined as P< 0.05. Results : The ADM group and rhBMP-2/ADM group results in enhanced local bone formation in the rat calvarial defect at both 2 and 8 weeks. The amount of defect closure and new bone formation were significantly greater in the rhBMP-2/ADM group relative to ADM group(P<0.05). At 8 weeks, the majority of ADM in the defect was contracted, and integrated with surrounding host tissues. In addition, host cell infiltration and neovascularization of the ADM in the absence of an inflammatory response were observed, and the newly formed bone around ADM showed a continuous remodeling and consolidation. Conclusion : The results of the present study indicated that ADM may be used as a barrier membrane for bone regeneration and that may be employed as a delivery system for BMPs.
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