不同种类烤瓷基底冠修复对基牙龈沟液及成分的影响

目的探讨不同种类烤瓷基底材料对基牙牙周组织的影响。方法选择上颌前牙烤瓷冠修复的患者48人,共72颗受试牙,按照烤瓷内冠不同分为镍铬合金、金合金和金沉积三组,每组24颗,患牙对侧同名牙为对照组。修复后6个月和12个月检测龈沟液(Gingival Crevicular Fluid,GCF)分泌量及其中肿瘤坏死因子-α(Tumour necrosis factor,TNF-α)和白细胞介素-6(Interleukin-6,IL-6)的含量。结果修复后6月镍铬合金和金合金组GCF分泌量及GCF中TNF-α含量明显高于对照组(P<0.05),IL-6含量升高无统计学意义(P>0.05)。12月后镍铬合金组和金合金组的GCF分泌量高于对照组(P<0.05),TNF-α和IL-6分泌量增加无统计学意义(P>0.05),金沉积组各指标水平无显著变化(P>0.05)。组间比较GCF量及TNF-α含量,镍铬合金组大于金合金组和金沉积组(P<0.05)。结论不同种类金属基底材料对牙周组织均有刺激性,但程度不同,长期效果有待继续观察。不同的牙科合金材料对龈沟液中某些细胞因子的含量有影响。     

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目的评价贵金属烤瓷全冠修复体对患牙牙周组织的影响。方法选择2007年4月至2008年12月于沈阳市沈河区牙病防治所行贵金属烤瓷全冠修复的患者25例(患牙77颗),于修复前和修复后6、12个月分别进行常规龈沟出血指数(SBI)检查、龈沟液(GCF)称量及GCF中白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)检测。结果肩台设计为龈下的前牙SBI在修复后12个月与修复前比较差异有统计学意义(P<0.05),GCF质量及GCF中IL-1β和TNF-α含量在修复前与修复后6个月、12个月比较差异无统计学意义(P>0.05);肩台设计为龈上的磨牙SBI、GCF质量及GCF中IL-1β和TNF-α含量在修复前与修复后6个月、12个月比较差异无统计学意义(P>0.05)。结论贵金属烤瓷全冠修复体对患牙的牙周组织影响较小,有利于牙周健康。     

Detection of hepatitis C virus RNA from gingival crevicular fluid and its relation to virus presence in saliva

BACKGROUND: To search for a possible source of hepatitis C virus (HCV) in saliva, the presence and shedding patterns of HCV in gingival crevicular fluid (GCF) and saliva of HCV viremic patients were assessed based on clinical, biochemical, histological, virological, and oral health parameters. METHODS: Saliva and GCF samples of 50 HCV viremic patients were collected to detect HCV RNA by a modified commercial polymerase chain reaction (PCR) assay. Clinical oral examination was performed and periodontal status at the collection sites was monitored. The results were correlated to specified parameters. RESULTS: HCV RNA was detected in 59% (29/49) of the GCF specimens and in 35% (17/48) of the saliva specimens. In saliva specimens, HCV RNA was detected only in cases which also had detectable HCV RNA in the GCF samples (P=0.00002) and was significantly related to the presence of blood in saliva (P=0.03). Higher, but not significant, values of oral clinical parameters at the sites of fluid collection were found in GCF specimens harboring HCV RNA. In GCF specimens with no blood detected, HCV RNA was more often present in cases with higher plasma viral load (P=0.05). CONCLUSIONS: The results suggest that besides blood, the other most probable source of HCV in saliva is GCF. Unknown endogenous HCV inhibitory mechanisms in the oral cavity may explain the discrepancies in HCV appearance between saliva and GCF. The results provide a biologic basis for further investigation of the role of HCV in the pathogenesis of periodontal disease.     

3种烤瓷全冠对龈沟液中TNF-α和IL-1β水平的影响

目的:探讨钴铬合金、纯钛金属及二氧化锆不同烤瓷冠基底材料对局部龈沟液中肿瘤坏死因子-α(TNF-α)以及白介素-1β(IL-1β)表达水平的影响。方法:选择临床病例30例,分为3组,分别采用钴铬合金烤瓷冠、纯钛烤瓷冠、二氧化锆全瓷冠修复。在修复前、修复后1个月、3个月分别进行龈沟液(GCF)量、菌斑指数(PLI)、TNF-α和IL-1β表达水平的测定及统计分析。结果:修复前3组之间各检测指标无明显差异;修复后1个月、3个月时钴铬合金烤瓷冠组的GCF量、TNF-α和IL-1β表达水平均高于纯钛烤瓷冠和二氧化锆全瓷冠组;而纯钛烤瓷冠组与二氧化锆全瓷冠组的GCF量、TNF-α和IL-1β水平无显著变化且两组之间各项指标无明显差异。结论:钴铬合金烤瓷冠可导致GCF量及其成分变化,对牙周组织有一定的不利影响,而纯钛烤瓷冠和二氧化锆全瓷冠对牙周组织健康的影响相对较小。     

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目的探讨钴铬合金烤瓷全冠修复对患牙牙周组织的影响。方法选择2007年4月至2008年10月来沈阳市沈河区牙病防治所应用钴铬合金烤瓷全冠修复的患者39例(106颗患牙),修复前和修复后6、12个月分别进行常规龈沟出血指数(SBI)检查,龈沟液(GCF)称量,GCF中白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)质量浓度的检测。结果修复前牙的患者患牙SBI、GCF质量、GCF中IL-1β和TNF-α的质量浓度修复前与修复后6个月比较差异无统计学意义(P>0.05),与修复后12个月比较差异有统计学意义(P<0.05);修复后牙的患者患牙SBI、GCF质量修复前与修复后6个月比较差异无统计学意义(P>0.05),与修复后12个月比较差异有统计学意义(P<0.05);修复后牙的患者患牙GCF中IL-1β和TNF-α的质量浓度修复前与修复后6个月和修复后12个月比较差异均无统计学意义(P>0.05)。结论采用钴铬合金烤瓷全冠修复前牙,修复时间超过12个月时,对SBI、GCF质量和GCF中IL-1β和TNF-α的质量浓度有显著影响;修复后牙,修复时间超过12个月时,对SBI、GCF质量有显著影响,而对GCF中IL-1β和TNF-α的质量浓度无明显影响。     

Correlations between pentraxin 3 or cytokine levels in gingival crevicular fluid and clinical parameters of chronic periodontitis

The gingival crevicular fluid (GCF) contains various biomarkers, such as interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor-α (TNF-α), and IL-10, among others. These cytokines have been reported to correlate with gingival inflammation and periodontal status. Therefore, the analysis of GCF may be useful for the diagnosis of periodontal status. Pentraxin 3 (PTX3) is the first identified long pentraxin, and is released by several cell types in response to proinflammatory signals. The aim of this study was to determine the levels of IL-1β, IL-6, IL-8, TNF-α, IL-10 and PTX3 in GCF from diseased and healthy sites in patients with chronic periodontitis. Cross-sectional clinical data were obtained from 50 patients with chronic periodontitis. GCF samples were collected with paper strips from one periodontal diseased site and one periodontally healthy site per subject. The levels of IL-1β, IL-6, IL-8, IL-10 and TNF-α were determined using a multiplexed bead immunoassay, and the PTX3 level was measured using an enzyme-linked immunosorbent assay. Mean clinical parameters were significantly higher at diseased sites (P < 0.01) as compared to healthy sites, and the mean levels of PTX3, IL-1β, IL-6, IL-8, IL-10 and TNF-α were higher in diseased sites (P < 0.01) than in healthy sites. There were strong correlations between PTX3 or IL-1β and periodontal status. These results suggest that GCF PTX3 levels might be useful as a diagnostic marker for periodontal disease.     

种植体周龈沟液高迁移率族蛋白B1的检测分析

目的:检测种植体周围炎患者龈沟液(gingival crevicular fluid,GCF)中高迁移率族蛋白B-1(high mobility groupbox l,HMGB-1)的水平,以探讨HMGB-1水平与种植体周围炎间的关系。方法 :经患者知情同意,选取行种植手术1年以上的健康种植体39颗和种植体周围炎40颗,共79颗种植体,收集种植体周围GCF,并运用ELISA法检测其中HMGB-1、IL-1β、IL-8、TNF-α的浓度,同时检测种植体周围的临床指标。结果 :轻度种植体周围炎组的HMGB-1、IL-1β、IL-8及TNF-α浓度高于健康种植体组,重度种植体周围炎的浓度水平又高于轻度种植体周围炎组,HMGB-1与IL-1β、TNF-α的浓度之间亦存在正相关关系(P<0.01)。结论:种植体周围GCF中HMGB-1水平变化与种植体周围炎的病变程度有关,且与经典炎症因子间存在显著相关性,临床检测GCF中HMGB-1水平可作为诊断种植体周围炎的客观指标。     

Localization of interleukin-1 (IL-1) mRNA-expressing macrophages in human inflamed gingiva and lL-1 activity in gingival crevicular fluid

The exact cell type and site(s) involved in interleukin-1 (lL-1) production during gingival inflammation was determined by combining immunohistochemistry and in situ hybridization. IL-1 messenger RNA (mRNA)-expressing cells in human inflamed gingiva were identified as macrophages. The rate of IL-α mRNA expression in these macrophages was the same as IL-1 β mRNA expression. The rate of IL-1 mRNA expression was higher in connective tissue furthest from the pocket epithelium, although more macrophages were present at the connective tissue subjacent to the pocket epithelium. The IL-1 activity in gingival crevicular fluid (GCF) obtained from inflamed gingiva was higher than that from healthy gingiva and decreased after periodontal therapy. The IL-1 activity in GCF was almost completely abolished by the addition of anti-IL-1α antibody but not by anti-IL-1 β antibody, indicating that IL-1α is the predominant form in GCF. However, the IL-1 activity in GCF was unrelated to the number of IL-1 mRNA-exprerssing macrophages in the same gingival site where the GCF was obtained at the same time. The results suggest that macrophages in the connective tissue subjacent to the oral epithelium contribute to the production of IL-1 but those in connective tissue subjacent to the pocket epithelium play a different role in the generation of gingival inflammation.     

Th2 cytokines efficiently stimulate periostin production in gingival fibroblasts but periostin does not induce an inflammatory response in gingival epithelial cells

Objectives

This study aims to clarify whether gingival fibroblasts produce periostin in response to Th2 cytokines which are elevated in periodontitis lesion and, if so, whether periostin affects the inflammatory response and matrix-protein metabolism.

Design

Human gingival fibroblasts, periodontal ligament cells and the gingival epithelial cell line epi4 were stimulated with interleukin-4 (IL-4), IL-13, tumour necrosis factor-α (TNF-α) and Porphyromonas gingivalis lipopolysaccharide (LPS). Periostin expression was analysed by real-time polymerase chain-reaction (PCR) and Western blotting. The expression of the IL-4 receptor α-chain was evaluated by immunocytochemistry. The effect of periostin on the production of inflammatory cytokines and the expression of matrix protein-related genes was analysed by real-time PCR and enzyme-linked immunosorbent assay (ELISA).

Results

While IL-4 and IL-13 significantly induced periostin production in gingival fibroblasts and periodontal ligament cells, no effect was observed in epi4 cells. No stimulatory effect of TNF-α or P. gingivalis LPS on the production of periostin was observed. The effect of periostin on the production of inflammatory cytokines was weak in gingival fibroblasts; however, little or no effect was observed on periodontal ligament cells or epi4 cells. No significant effect of periostin on the expression of matrix protein-related genes was found.

Conclusion

The results suggest that gingival fibroblasts may be a source of periostin in periodontitis lesions but periostin has only a limited role either in the inflammatory response or in matrix-protein metabolism. Thus, the role of periostin in the cellular interaction between epithelial and mesenchymal cells in gingiva may be distinct from that of skin.     

Concentrations of interleukin-32, interleukin −10, interleukin −6, and TNF-alfa are higher in saliva of children with early childhood caries

AimTo investigate if the levels of interleukin-32 (IL-32), interleukin-10 (IL-10), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α) in the saliva are associated with early childhood caries.MethodsA total of 56 patients aged between 36 and 71 months with dental caries and without caries were included in this study. The patients’ caries status was evaluated according to the dmft and dmfs indices. IL-32, IL-10, IL-6, and TNF-α levels in the saliva samples of the patients were measured by ELISA.ResultsThere were no statistically significant differences in the oral hygiene and nutritional habits, plaque index, and gingival index values between the ECC and control groups evaluated in the study (p > 0.05). The ECC group's salivary IL-32, IL-10, IL-6, and TNF-α levels were statistically significantly higher than the control group (p < 0.001).ConclusionThe high levels of IL-32, IL-10, IL-6, and TNF-α detected in the saliva of children with ECC reveal that these cytokines may play a potential role in ECC pathogenesis. Salivary levels of IL-32, IL-10, IL-6, and TNF-α are associated with early childhood caries in children.     

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摘要：目的    分析采用氧化锆全瓷冠修复对患牙牙周组织的影响。方法    选择2008 年 6月至2009年3月在沈阳市口腔医院修复科就诊的需行全冠修复的患者38例（患牙52颗）,采用氧化锆全瓷冠修复患牙。于修复前及修复后1年行牙周状况检查。分别取患牙修复前、后的龈沟液（gingival crevicular fluid,GCF）,进行GCF量测定,并对GCF内肿瘤坏死因子（tumor necrosis factor-α,TNF-α）和白细胞介素-6（interleukin-6,IL-6）含量进行比较分析。结果    修复体边缘的深度符合临床要求,修复后患牙的牙周指标（探诊深度、龈沟出血指数、菌斑指数）、GCF量、GCF内TNF-α及IL-6含量与修复前比较,差异无统计学意义（P > 0.05）。结论    氧化锆全瓷冠修复后1年内对患牙牙周组织无明显不利影响。     

Expression of HMGB1 and HMGN2 in gingival tissues, GCF and PICF of periodontitis patients and peri-implantitis

Background and objective

High mobility group chromosomal protein B1 (HMGB1) and N2 (HMGN2), two members of high mobility group (HMG) family, play important role in inflammation. The purpose of this study was to investigate the expression of HMGB1 and HMGN2 in periodontistis.

Materials and methods

The expression of HMGB1 and HMGN2 mRNA in gingival tissues and gingival crevicular fluid (GCF) in chronic periodontitis (CP), generalised aggressive periodontitis (G-AgP) patients and healthy subjects was detected by real-time PCR. The protein level of HMGB1 and HMGN2 in peri-implant crevicular fluid (PICF), peri-implant crevicular fluid of peri-implantitis (PI-PICF) and normal patients was determined by Western blotting. Furthermore, IL-1β, IL-6, IL-8, TNF-α and HMGB1 levels in GCF, PI-PICF and healthy-PICF samples from different groups were determined by ELISA.

Results

HMGN2 expression was increased in inflamed gingival tissues and GCF from CP and G-ApG groups compared to control group. HMGB1 expression was the highest in the gingival tissues and GCF from CP patients and was accompanied by increased concentrations of IL-1β, IL-6, IL-8 proinflammaory cytokines.

Conclusion

To our knowledge, this is the first study reporting that the expression of HMGB1 and HMGN2 was increased in the gingival tissues and GCF in CP and G-AgP and the PICF in PICF. Our data suggest that HMGB1 may be a potential target for the therapy of periodontitis and PI.     

Expression of secretory proteins in oral fluid after orthodontic tooth movement.

Orthodontic treatment alters the expression of secretory proteins at the local level in bone and the oral cavity, but its systemic effects are not well understood. Total secretory proteins and a cyclic adenosine monophosphate (AMP)-dependent protein kinase subunit (RII) were measured in saliva and gingival crevicular fluid (GCF) after the placement of orthodontic separators to determine if mechanical force applied to teeth affects protein secretion. Whole saliva and GCF were collected before and 1 day after treatment. Electrophoresis and Western blotting were carried out to establish the banding patterns of total proteins and to measure the isotype and amount of RII that serves as an apparent stress indicator. Digitized image files were used for densitometric analyses of the relative concentrations of RII and total protein. Individual protein values showed no statistically significant changes in saliva or GCF. Western blots, however, showed a dramatic difference in RII after the placement of separators: the 50-to-55 kilodaltons (kd) band virtually disappeared and was replaced by a fragment in the 20-kd range. These results suggest that although the expression of total proteins is not altered by mechanical force applied to teeth, a systemic response via the cyclic AMP signaling pathway might have been activated.     

白藜芦醇对高糖环境下牙龈上皮细胞TLR4表达及炎症因子分泌的影响

目的:研究白藜芦醇对高糖环境下牙龈上皮细胞TLR4表达及炎症因子分泌的影响,探讨白藜芦醇对伴糖尿病牙周炎患者的治疗作用及相关分子机制.方法:体外培养牙龈上皮细胞,按照作用方式不同分为正常对照组、高糖组和高糖+白藜芦醇组.荧光定量PCR检测TLR4表达情况;取第3代牙龈上皮细胞,高糖环境下采用或不采用白藜芦醇处理24 h,随后用100 ng/mL LPS处理2 h,ELISA检测IL-1β 、IL-6、IL-8和TNF-α分泌;Western印迹法检测TLR4信号通路下游分子NF-κB p65、p38MAPK和STAT3磷酸化.采用SPSS 17.0软件包对数据进行统计学分析.结果:白藜芦醇可以逆转高糖环境下牙龈上皮细胞TLR4水平的升高,同时抑制高糖环境下LPS诱导的IL-1β、IL-6、IL-8和TNF-α表达.Western印迹结果显示,白藜芦醇也可抑制TLR4信号通路下游分子NF-κB p65、p38MAPK和STAT3磷酸化.结论:白藜芦醇通过负向调控TLR4信号通路减轻高糖环境下牙龈上皮细胞炎症因子的分泌.     

Ghrelin levels in chronic periodontitis patients

Ghrelin is a peptide hormone that has modulatory effects on the immune system. This study was designed to evaluate plasma ghrelin levels in patients with chronic periodontitis and to investigate if a relationship exists between ghrelin and periodontal parameters, serum cytokines, and bone turnover markers. Thirty-five chronic periodontitis patients (CP) and periodontal healthy individuals (C) were included in this study. Periodontal parameters were recorded. Blood samples were obtained to determine the levels of total and acylated ghrelin, interleukin-1 beta (IL-1β), tumor necrosis factor-alpha (TNF-α), the soluble receptor activator nuclear factor kappaB ligand (sRANKL), alkaline phosphatase (ALP), and osteocalcin (OSC). Plasma levels of total and acylated ghrelin were significantly elevated in the CP group compared with the C group (p < 0.05). The difference was significant only between males in the two groups (groups were compared with respect to gender) (p < 0.05). There was no difference between the groups regarding the levels of serum sRANKL, TNF-α, and ALP. A relative increase in the serum levels of IL-1β and a decrease in the serum levels of OSC of the CP group were observed (p < 0.05). In addition, positive correlations between total ghrelin/ALP and total ghrelin/acylated ghrelin were discovered. We found no direct correlation between ghrelin levels and periodontal parameters. Our results indicate an increase of total and acylated ghrelin levels in patients with chronic periodontitis. Further, studies in larger populations (which could include ghrelin levels in gingival tissue, gingival crevicular fluid, and saliva) are needed in order to confirm the role of ghrelin in periodontal disease.     

Aggregatibacter actinomycetemcomitans lipopolysaccharide stimulated epithelial cells produce interleukin-15 that regulates T cell activation

ObjectiveOral epithelial cells act not only as mechanical barriers but also as immunological barriers by producing various mediators such as cytokines. Since, in periodontal disease, limited information is available regarding the role of oral epithelial cell-derived cytokines on T cell activation, we investigated the responses of human T cells (Jurkat cell) to cytokines in KB cells (an oral epithelial cell line) that had been stimulated with Aggregatibacter actinomycetemcomitans lipopolysaccharide (LPS).DesignTo evaluate T cell activation in response to the culture supernatant of KB cells, we examined cell proliferation and interferon gamma (IFN-γ) production, which is closely related to periodontal disease, in Jurkat cells. Culture supernatant of LPS-stimulated KB cells enhanced cell proliferation and IFN-γ production in Jurkat cells. To determine the active component within the culture supernatant, the production of epithelial cell-derived cytokines, interleukin-12 (IL-12), IL-15 and IL-18, in LPS-stimulated KB cells was analysed.ResultsIL-15, but not IL-18, was significantly increased in the culture supernatant of LPS-stimulated KB cells. Moreover, additional anti-IL-15 neutralizing antibody abolished culture supernatant-induced IFN-γ expression in Jurkat cells.ConclusionThese results suggest that periodontal pathogens induce the production of IL-15 from epithelial cells, and leading the activation of T cells in periodontal lesions.     

The evaluation of cystatin C, IL-1beta, and TNF-alpha levels in total saliva and gingival crevicular fluid from 11- to 16-year-old children

BACKGROUND: The aim of this study was to evaluate the levels of cystatin C, interleukin-1beta (IL-1beta), and tumor necrosis factor-alpha (TNF-alpha) in the total saliva and gingival crevicular fluid (GCF) of periodontally healthy children (PHC) and children with gingivitis (CG) who were between 11 and 16 years old. METHODS: The study was carried out with 10 PHC and 25 CG. Unstimulated total saliva and GCF samples were obtained. Clinical parameters, including probing depth (PD), clinical attachment loss (CAL), plaque index (PI), gingival index (GI), and gingival bleeding index (GBI), were assessed. GCF samples were collected from four maxillary upper incisors. After sampling, biochemical analyses were performed using latex particle-enhanced turbidimetric immunoassay for cystatin C and enzyme-linked immunosorbent assay for IL-1beta and TNF-alpha. The multivariate analysis of variance test was used for statistical evaluation. RESULTS: In total saliva, cystatin C and TNF-alpha levels were higher in PHC, and IL-1beta levels were higher in CG, but the differences were not statistically significant. In GCF, cystatin C levels were higher in PHC (P >0.05), whereas TNF-alpha and IL-1beta levels were higher in CG (P >0.05). In the CG group, there were positive correlations between the GCF cystatin C level and the PI of the sampled site (r = 0.488; P <0.05); also, GCF IL-1beta (r = 0.603; P <0.05) and TNF-alpha (r = 0.456; P <0.05) levels were positively correlated with PD and CAL. For the whole mouth and the sampled sites, PI, GI, GBI, PD, and CAL values were higher in CG (P <0.05), but no significant differences were detected between GCF volumes of the two groups. CONCLUSIONS: To the best of our knowledge, this study represents the first evaluation of cystatin C in the gingival disease mechanism in children. Our results showed that total saliva and GCF cystatin C levels were higher in PHC (P >0.05), but there was no correlation between cystatin C levels and IL-1beta or TNF-alpha levels in total saliva or GCF.