程序性死亡配体1负性调控慢性牙周炎的炎症反应

目的 探讨程序性死亡配体1（PD-L1）在慢性牙周炎中是否有表达,以及与不同程度慢性牙周炎的相关关系,为阐明慢性牙周炎的免疫调控机制、临床治疗和预后提供依据。方法 收集健康人和慢性牙周炎患者的牙龈和牙周膜组织。根据临床探测,分为正常对照组、轻度牙周炎组、重度牙周炎组。运用荧光定量聚合酶链反应（PCR）测定不同组牙周组织中PD-L1 mRNA的表达;免疫印迹（Western blot）以及免疫组织化学方法测定不同组牙周组织中PD-L1蛋白的表达。结合临床影像资料,分析PD-L1的差异表达量和不同程度慢性牙周炎的关系。结果 轻度牙周炎组牙周组织中PD-L1的相对表达量显著高于重度牙周炎组（P<0.01）;正常对照组牙周组织中PD-L1的相对表达量与重度牙周炎组牙周组织相比无统计学意义（P>0.05）。结论 PD-L1在牙周组织的表达可负性调控炎症牙周组织损伤。     

糖基化终末产物受体对牙周炎的促进作用

晚期糖基化终末产物受体（RAGE）是一种多配体受体,它通过与相应的配体结合后启动若干信号通路,影响机体的免疫防御和创伤愈合,参与炎症的恶性循环。在牙周组织中,RAGE在病理状态下的表达增加,提示该受体可能是牙周炎的促进因子之一,并有可能参与糖尿病和吸烟时牙周组织的损伤。下面就RAGE及其在牙周慢性炎症中的作用和在牙周组织中的表达,以及RAGE配体在牙周组织中的作用等作一综述。     

β-防御素-3及其与牙周炎和糖尿病的关系

β-防御素-3的表达与牙周炎和糖尿病密切相关,而后两者间亦关系密切。研究显示,β-防御素-3在牙周炎患者龈沟液中的表达明显降低;且当炎症越重,致病菌种类越多时,β-防御素-3的表达越低。格氏链球菌、血链球菌和非典型韦荣菌等牙周非致病菌轻微上调β-防御素-3的表达,具核梭杆菌和中间普雷沃菌等橙色复合体细菌可强烈地诱导β-防御素-3分泌,齿垢密螺旋体、牙龈卟啉单胞菌和福赛斯拟杆菌等红色复合体细菌则会抑制β-防御素-3的产生。β-防御素-3在健康牙周组织中的表达相对于炎症牙周组织较高,有利于维持牙周组织的稳定。β-防御素-3与牙周有关的生物学功能主要包括抗菌活性、免疫调节作用和促进细胞增殖生长等。在糖尿病状态下,机体的免疫防御功能受损,β-防御素-3在糖尿病感染伤口组织表达降低。目前,有关β-防御素-3的可能的不良反应、生物学功能、在感染性疾病中的具体作用机制还有待进一步研究。     

β-防御素2在实验性大鼠牙周炎口腔龈上皮、颊黏膜上皮及腮腺导管上皮的表达

目的:探讨β-防御素2在大鼠牙周炎模型口腔龈上皮、颊黏膜上皮和腮腺导管上皮的表达变化及与牙周炎发展的关系。方法:利用结扎法建立大鼠上颌第一磨牙牙周炎模型并设对照组,于第6周取实验牙及牙周组织、相对颊黏膜和腮腺组织,制备组织切片,免疫组化染色法检测防御素2在口腔龈上皮、颊黏膜上皮、腮腺导管上皮的表达并行免疫组化评分和统计学分析。结果:β-防御素2主要表达于口腔龈上皮和颊黏膜上皮的棘层和颗粒层以及腮腺的导管上皮细胞。在口腔龈上皮的表达为牙周炎组显著低于对照组(P<0.01),在颊黏膜上皮的表达为牙周炎组显著高于对照组(P<0.01),而在腮腺导管上皮的表达牙周炎组和对照组无显著差异。结论:牙周炎症时β-防御素2在口腔龈上皮、颊黏膜上皮和腮腺导管上皮的分布各有不同的变化,提示在牙周炎进展过程中不同部位口腔上皮对牙周化学屏障改变的影响不同。     

人β-防御素-3对小鼠牙周炎进展的保护作用

目的:探索人β-防御素-3在小鼠牙周炎进展中的保护作用。方法:用丝线结扎法建立小鼠牙周炎模型。实验组小鼠腹腔注射人β-防御素-3(100 μg/kg),另选结扎过小鼠和未结扎小鼠各1组,腹腔注射磷酸盐缓冲液为对照。8周后取上颌骨,Micro CT、HE染色、TRAP染色及免疫组化染色,评估人β-防御素-3对牙周炎进展的作用。结果:人β-防御素-3能减少牙槽骨吸收及骨内破骨细胞数量;减轻牙龈上皮棘层增厚、上皮钉突增生、炎症细胞浸润、胶原纤维变性断裂等病理变化,降低牙周组织中IL-6、MCP-1和TNF-α的表达强度。结论:人β-防御素-3能够减轻牙周组织炎症,减少牙周组织破坏,从而减缓牙周炎进展。     

慢性牙周炎中核因子κB受体活化子、护骨素和肿瘤坏死因子-α的蛋白表达

目的　比较慢性牙周炎牙槽骨吸收处肉芽组织中的核因子κB受体活化子 (RANKL)、护骨素 (OPG)和肿瘤坏死因子(TNF) -α的水平 ,研究慢性牙周炎牙槽骨吸收与这些调节因子间的关系。方法　应用鼠抗人单克隆抗体、半定量分析及数字成像技术分析慢性牙周炎组织中RANKL、OPG和TNF α的蛋白表达。结果　慢性牙周炎牙槽骨吸收处肉芽组织中有较高的RANKL和TNF α蛋白表达 ,OPG蛋白表达相对较低 (P <0 .0 5 ) ;非牙周炎组织中显示较强的OPG蛋白表达和较弱的RANKL和TNF -α蛋白水平 (P <0 .0 5 ) ;OPG蛋白表达与牙周组织中内皮细胞相关。结论　RANKL可能与慢性牙周炎牙槽骨吸收相关 ;RANKL和OPG平衡的变化在慢性牙周炎骨吸收的分子机制中起重要调节作用 ;TNF α可能协同参与牙槽骨的吸收     

内质网应激诱导的细胞凋亡在慢性牙周炎中的作用初探

目的:初步探讨内质网应激诱导的细胞凋亡在慢性牙周炎中的作用。方法:利用光学显微镜和透射电镜观察牙周组织中的凋亡细胞,免疫组化染色法检测慢性牙周炎患者及牙周健康者牙周组织中GRP78和CHOP的分布及含量变化。结果:慢性牙周炎组内浆细胞呈现凋亡状态,且GRP78和CHOP在慢性牙周炎组中的阳性表达明显高于正常组(P<0.05)。结论:内质网应激参与了慢性牙周炎的病理生理过程。     

氧化应激及抗氧化防御系统在慢性牙周炎中的作用

近年来研究发现,氧化应激与抗氧化防御系统的失衡在慢性牙周炎中发挥重要作用,牙周组织受到细菌刺激产生的过量活性氧可造成和加重牙周组织的损伤。本文就炎性条件下过量活性氧对牙周组织的损害,以及氧化应激与抗氧化防御系统在慢性牙周炎发生发展中的作用和机制作一综述。     

龈沟液β-防御素-2、β-防御素-3在慢性牙周炎与2型糖尿病中的水平研究

目的通过检测龈沟液中β-防御素2、β-防御素3的表达水平,探讨其在慢性牙周病与2型糖尿病中的可能作用。方法选取慢性牙周炎伴2型糖尿病组(T2DMCP组)20例,2型糖尿病组(T2DM组)20例,慢性牙周炎组(CP组)20例,健康组(H组)20例作。采用酶联免疫吸附试验(ELISA)检测龈沟液中β-防御素-2和β-防御素-3水平。比较组间龈沟液中β-防御素-2和β-防御素-3水平的差异性,分析β-防御素-2和β-防御素-3之间的相关性与β-防御素-2和β-防御素-3与牙周指标的相关性。结果 CP组、T2DM组、T2DMCP组的临床指标PD、CAL、BI和GCF量均高于H组(P<0.05)。T2DMCP组的PD、CAL、BI和GCF量均高于T2DM组和CP组(P<0.05)。龈沟液中β-防御素-2,β-防御素-3水平在H组高于CP组、T2DM组和T2DMCP组(P<0.05),CP组和T2DM组高于T2DMCP组(P<0.05)。龈沟液中β-防御素-2水平与β-防御素-3水平呈正相关(P<0.05)。结论慢性牙周炎和2型糖尿病可降低龈沟液β-防御素-2和β-防御素-3的水平,提示β-防御素-2、β-防御素-3对牙周组织起保护作用。     

正畸力影响炎性牙周组织中肿瘤坏死因子α蛋白表达的实验研究

目的　探讨正畸力及炎性刺激影响牙周组织改建的联合效应。方法　选用96只雄性SD成年大鼠,分别 建立实验性牙周炎大鼠和注射内毒素健康大鼠模型,近中移动两模型的上颌磨牙。结果　在0 h、2 h、12 h、2 d、7 d、 14 d时,磨牙近中压力侧牙周膜组织内肿瘤坏死因子α(TNF-α)的蛋白表达出现波动,2 d时平均光密度(OD)值达到 峰值。结论　急、慢性牙周炎症均可阻碍正畸力效应下的牙周组织改建,研究提示对成年牙周炎患者,牙移动应谨 慎进行。     

The expression of hBDs in the gingival tissue and keratinocytes from healthy subjects and periodontitis patients

ObjectiveAlthough the secretion of antimicrobial peptides in gingival tissue and isolated cells has been reported, the induction of human β-defensins (hBDs) in epithelial cells from the periodontitis patients was not stated before. This study aimed to compare the secretion of hBDs in gingival epithelial cells from periodontitis patients and healthy controls.DesignFirstly, gingival biopsies were obtained from chronic periodontitis patients and healthy controls and the hBDs expression level in gingival tissues was quantified. Then the epithelial cells from periodontitis patients and healthy controls were isolated and challenged with different concentrations of tumour necrosis factor-alpha (TNFα). The hBDs expression level was also quantified after induction. At last, to identify the molecular pathways involved in hBDs induction, the isolated cells were incubated with NF-kB or MAPK inhibitor before TNFα induction.ResultsHigher hBDs expression was found in gingival tissues from healthy controls. The in vitro experiments demonstrated that the hBD-2 expression in gingival epithelial cells from periodontitis patients can be induced by TNFα at lower dose, while the optimum expression level was much lower. The basal hBD-3 mRNA expression was much higher in cells from periodontitis patients. The molecular pathways involved in the responses to the inflammatory cytokine in patients and healthy controls were the same.ConclusionsThe epithelial cells from periodontitis patients are more prone to recognize and respond to TNFα to produce hBD-2. The basal expression of hBD-3 in keratinocytes from periodontitis patients suggested that hBD-3 may play an important role in the immunological reaction against periodontitis.     

Expression of toll-like receptors 2, 4 and 9 is increased in gingival tissue from patients with type 2 diabetes and chronic periodontitis

Rojo‐Botello NR, García‐Hernández AL, Moreno‐Fierros L. Expression of toll‐like receptors 2, 4 and 9 is increased in gingival tissue from patients with type 2 diabetes and chronic periodontitis. J Periodont Res 2012; 47: 62–73. © 2011 John Wiley & Sons A/S Background and Objective: Broad evidence indicates that diabetes both increases the risk and hastens the progression of periodontal disease. Likewise, chronic inflammation or infections seem to provoke insulin resistance and thereby contribute to the development of diabetes and its complications. Innate immune responses, which appear to be altered in individuals with diabetes, are usually mediated by the recognition of pathogens through toll‐like receptors (TLRs). The constitutive expression of some TLRs has been reported in healthy human gingival tissue. Interestingly, the expression of TLRs 2 and 4 is increased with the severity of periodontal disease. Considering that the inflammatory reaction is exacerbated in individuals with diabetes and periodontitis, we suspected that the expression of some TLRs might be increased in gingival tissue in these patients. Material and Methods: In this study, we analyzed, by immunofluorescence, the expression of TLRs 2, 3, 4 and 9 in gingival tissues from healthy individuals and from periodontal patients with or without type 2 diabetes. Results: We found that the expression levels of TLRs 2, 3, 4 and 9 were higher in all periodontal patients than in healthy individuals. The expression of some TLRs was increased in subjects with periodontitis and diabetes relative to subjects with periodontitis but without diabetes; this increase in expression was found particularly in TLR2 and TLR9 in the connective tissue and in TLR4 at the epithelial region. Conclusion: These data suggest that the expression of these TLRs 2, 3, 4 and 9 in gingival tissue is higher in individuals with diabetes because its inflammatory reaction is exacerbated. Additionally, the expression of these TLRS is positively regulated with the severity of periodontal disease.     

巨噬细胞移动抑制因子在慢性牙周炎患者血清及牙龈组织中的表达分析

目的    观察并分析慢性牙周炎患者血清及牙龈组织中巨噬细胞移动抑制因子（macrophage migration inhibitory factor,MIF）表达及其临床意义。方法    选择2016年6月至2017年6月就诊于中国医科大学附属口腔医院牙周科及口腔外科的患者,收集重度慢性牙周炎患者（牙周炎组,18例）及牙周健康者（健康对照组,18例）血清,酶联免疫吸附试验检测MIF及单核细胞趋化蛋白-1（monocyte chemotactic protein-1,MCP-1）水平。收集未经治疗（CP组,8例）及牙周基础治疗后1个月（SRP组,11例）的重度慢性牙周炎患者、牙周健康者（H组,18例）的牙龈组织,采用免疫组化法检测MIF及MCP-1在牙龈中的表达。采用SPSS 24.0软件t检验比较血清中MIF及MCP-1表达的差异,单因素方差分析比较CP组、SRP组及H组牙龈组织中MIF及MCP-1表达的差异。结果    慢性牙周炎患者血清中MIF及MCP-1浓度较牙周健康者明显升高（P < 0.05）。MIF在牙龈上皮呈阳性表达,且在未经治疗的CP组表达水平最高,经牙周基础治疗后的SRP组MIF表达水平显著下降,但仍高于H组（P < 0.05）;MCP-1在牙龈组织的上皮层和结缔组织中均有弱表达,表达水平在3组间差异无统计学意义（P > 0.05）。结论    MIF作为一种炎症因子参与慢性牙周炎的发展进程,其表达水平可作为炎症指标辅助慢性牙周炎活动性及炎症程度的判断。     

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??Objective    To investigate the expression of macrophage migration inhibitory factor??MIF??in the serum and gingival tissues of patients with chronic periodontitis??and to explore its role in clinical prognosis. Methods    Serum samples were collected from chronic periodontitis patients??18 cases??and healthy controls??18 cases????MIF and monocyte chemotactic protein-1??MCP-1??levels were assayed by enzyme-linked immunosorbent assay. Gingival tissues samples were collected from chronic periodontitis patients??before and after periodontal initial therapy??and periodontally healthy individuals. Then the MIF and MCP-1 expression were detected with immunohistochemical method. The data was analyzed with SPSS 24.0 software package. Results     Serum MIF and MCP-1 were significantly higher in periodontitis patients compared with healthy controls??P < 0.05??. MIF protein was expressed in gingival epithelia??and the MIF level was highest in periodontitis gingiva??its level was decreased by periodontal initial therapy??but still higher than the expression of MIF in healthy gingiva ??P < 0.05????MCP-1 protein was also expressed in both gingival epithelia and connective tissues??but there was no significant difference between the periodontitis patients and healthy controls??P > 0.05??. Conclusion          MIF is involved in the progression of chronic periodontitis as an inflammatory factor??and its level may be used as auxiliary indicators for the evaluation of the activity and degree of periodontitis.     

Salivary and plasma levels of Toll-like receptor 2 and Toll-like receptor 4 in chronic periodontitis

Background: This cross‐sectional study was planned to investigate whether patients with chronic periodontitis exhibit different salivary or plasma concentrations of Toll‐like receptor (TLR) 2 and TLR4 compared to subjects who are clinically healthy. Methods: Whole saliva and plasma samples were obtained and full‐mouth clinical periodontal measurements were recorded from 22 otherwise healthy patients with chronic periodontitis and 21 systemically and periodontally healthy control subjects. Salivary and plasma TLR2 and TLR4 levels were determined by enzyme‐linked immunoassays. Data were tested statistically using Mann‐Whitney U test. Results: The healthy group exhibited significantly lower values in all clinical measurements (P <0.001). The salivary TLR2 levels were similar in the two study groups (P >0.05). The patients with chronic periodontitis exhibited significantly higher salivary TLR4 (P <0.01) and plasma TLR2 and TLR4 levels (P <0.05). Conclusion: The present findings support a hypothesis that inflammation increases expression of TLRs which leads to an increased detection of TLRs in saliva and plasma, which could be useful as a diagnostic test for periodontal diseases.     

Innate Immune Receptor Expression in Peri‐Implant Tissues of Patients With Different Susceptibility to Periodontal Diseases

Background: Although inflammation mediates the pathogenesis of periodontal diseases, the effects of innate immune responses on implant therapies have not been evaluated. Innate immune receptors, including toll‐like‐receptors (TLRs) and the receptor for advanced glycated end‐products (RAGE), are upregulated within inflamed gingiva and are responsible for initiation of detrimental host responses. The aim of this study is to compare the expression of TLR2, TLR4, and RAGE in gingival tissues from participants susceptible to periodontitis and participants not susceptible to periodontitis before and after implant therapy. Methods: Periodontally healthy participants received implant therapy for non‐periodontal edentulism. Participants susceptible to periodontitis were diagnosed with chronic periodontitis prior to implant therapy. Gingival biopsies were collected from edentulous ridges before implant installation and from peri‐implant mucosa 2 months after treatment. Histology, real‐time PCR, and Western blot were used to evaluate levels of inflammatory infiltrate, TLR2, TLR4, and RAGE expression. Results: Before implant therapy, elevated levels of RAGE were detected in gingival tissues from participants susceptible to periodontitis when compared to those from participants with healthy periodontiums, whereas no differences in the expression of TLR2 or TLR4 were detected. After implant therapy, there was an upregulation of RAGE and TLR4 levels that coincided with a downregulation of TLR2 levels in biopsies from participants susceptible to periodontitis. Levels of RAGE and TLR4 remained unchanged in biopsies from participants with healthy periodontiums, whereas TLR2 levels were significantly upregulated. Histologically, post‐implant biopsies from participants susceptible to periodontitis displayed higher levels of inflammatory infiltrate. Conclusion: Elevated levels of inflammatory potential were found after implant therapy in participants susceptible to periodontitis.     

Expression of human beta-defensins-1 and -2 peptides in unresolved chronic periodontitis

BACKGROUND: Human beta-defensins (hBDs) are antimicrobial peptides which contribute to host innate immunity by disrupting the membrane integrity of a broad spectrum of microorganisms. OBJECTIVES: This study aimed to determine the expression profiles of hBD-1 and -2 peptides in gingiva and to assess the possible relations of these antimicrobial peptides with periodontal health and disease. METHODS: Seven periodontally healthy subjects and 22 patients with unresolved chronic periodontitis were recruited and the gingival biopsies collected consisted of healthy tissues from the healthy subjects (HT-C); periodontal pocket tissues (PoT) and inflamed connective tissues (ICT) from the base of pocket, i.e. granulation tissues, as well as clinically healthy tissues (HT-P) from the adjacent clinically healthy sites from the patients. The expression of hBD-1 and -2 peptides was detected by immunohistochemistry and quantitatively analyzed with a computerized image processing system. RESULTS: Both hBD-1 and -2 peptides were detected in all periodontally healthy subjects, while hBD-1 was detected in all patients and hBD-2 was found in most of the patients. Their expression was mainly confined to the granular and spinous layers of gingival epithelium, in which hBD-1 was detected in both intercellular spaces and cytoplasm, whereas hBD-2 was mainly observed in the cytoplasm. HT-C expressed significantly higher levels of hBD-2 than HT-P (p < 0.05). Within the patients, both defensins were up-regulated significantly in PoT as compared with the adjacent HT-P (p < 0.05). CONCLUSIONS: The present study showed that hBD-1 and -2 were frequently expressed in the granular and spinous layers of gingival epithelia and their expression may be associated with periodontal health and disease.     

Immunohistochemical localization of Toll‐like receptors 1–10 in periodontitis

Background/aim: In periodontitis, bacteria and pathogen‐associated molecular patterns are sensed by Toll‐like receptors (TLRs), which initiate intracellular signaling cascades that may lead to host inflammation. In this study, the expression and distribution of TLRs (TLR‐1 to TLR‐10) were immunohistochemically detected in gingival epithelium and connective tissue. Methods: Immunohistochemistry was used for the localization of TLRs in gingival tissue samples from 10 patients with chronic periodontitis and 10 healthy controls; these samples were obtained during routine periodontal flap operations and during extraction operations performed for retained wisdom teeth, respectively. For the evaluation, epithelial layers were stratified to basal, spinous, and superficial layers, and the percentages of TLR‐positive cells were determined. Results: Both healthy and periodontitis gingival tissues expressed all TLRs except TLR‐10. In patients with periodontitis, epithelial cells showed increased TLR expression towards the basal layer. Healthy controls showed more variable cellular TLR expression and distribution between the layers of epithelium. In the connective tissue, consistently higher TLR expression was found within the periodontitis group compared to the healthy group. Conclusions: For the first time, the cellular expression and distribution of TLR‐1 to TLR‐10 have been studied in periodontitis, indicating that TLR‐1 to TLR‐9 are differentially expressed both in connective tissue and epithelial layers. Except for TLR‐7 and TLR‐8, all the other TLRs showed statistically significant differences between patients with periodontitis and healthy controls, suggesting their involvement in the pathogenesis of periodontitis.     

Immunohistochemical localization of Toll-like receptors 1-10 in periodontitis

Background/aim:  In periodontitis, bacteria and pathogen-associated molecular patterns are sensed by Toll-like receptors (TLRs), which initiate intracellular signaling cascades that may lead to host inflammation. In this study, the expression and distribution of TLRs (TLR-1 to TLR-10) were immunohistochemically detected in gingival epithelium and connective tissue.
Methods:  Immunohistochemistry was used for the localization of TLRs in gingival tissue samples from 10 patients with chronic periodontitis and 10 healthy controls; these samples were obtained during routine periodontal flap operations and during extraction operations performed for retained wisdom teeth, respectively. For the evaluation, epithelial layers were stratified to basal, spinous, and superficial layers, and the percentages of TLR-positive cells were determined.
Results:  Both healthy and periodontitis gingival tissues expressed all TLRs except TLR-10. In patients with periodontitis, epithelial cells showed increased TLR expression towards the basal layer. Healthy controls showed more variable cellular TLR expression and distribution between the layers of epithelium. In the connective tissue, consistently higher TLR expression was found within the periodontitis group compared to the healthy group.
Conclusions:  For the first time, the cellular expression and distribution of TLR-1 to TLR-10 have been studied in periodontitis, indicating that TLR-1 to TLR-9 are differentially expressed both in connective tissue and epithelial layers. Except for TLR-7 and TLR-8, all the other TLRs showed statistically significant differences between patients with periodontitis and healthy controls, suggesting their involvement in the pathogenesis of periodontitis.