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.     

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.     

Recurrent aphthous ulcers—a Toll‐like receptor–mediated disease?

J Oral Pathol Med (2012) 41 : 158–164 Background: Recurrent aphthous ulcer (RAU) is characterized by acute and painful inflammatory ulcerations, which heal spontaneously but tend to recur. Many pathogens have been proposed as causative agents, but none has been consistently proven. According to our hypothesis, RAU is an autoinflammatory disorder triggered by pathogen‐associated molecular patterns (PAMPs) shared by different pathogenic and commensal microbes. Methods: PAMP‐reactive Toll‐like receptors (TLRs) were mapped in oral epithelium in healthy controls compared to RAU. Results: In controls, the superficial epithelium formed a TLR^?, a PAMP non‐reactive physical barrier zone, but all TLRs were found deeper in the epithelium, usually restricted to suprabasal and basal cell layers. In RAU, the epithelial TLR polarity was lost: TLRs 1, 2, 5, 7, and 8 were found throughout the epithelium, but also TLRs 4, 6, and 10 extended higher up than normally, whereas TLR‐3 was almost lost in RAU. In RAU lesions, connective tissue stroma was heavily infiltrated by TLR⁺ inflammatory cells. Conclusions: Normal TLR architecture prevents inflammatory responses against normal microbes but still contains a deep TLR⁺, PAMP‐reactive dormant defense zone. In RAU, the TLR⁺, PAMP‐reactive zone extends to surface or subsurface exposed to microbial PAMPs. TLR reactivity is further enhanced by recruitment of inflammatory leukocytes forming a new deep line of defense. The organization of the TLR system in healthy mucosa and its changes in RAU are compatible with active pathogenic involvement of TLRs, which together with the typical clinical picture and course suggest that RAU is a TLR‐mediated disease.     

Differential expression of Toll‐like receptors in chronic hyperplastic candidosis

Introduction: The first step in the host defense against oral candidosis is the recognition of Candida albicans through a set of germ‐encoded pathogen recognition receptors, e.g. Toll‐like receptors (TLRs). In man, 10 types of such receptors have been identified so far, of which only TLR2, TLR4, and TLR6 have been linked to mediating candidal ligands, e.g. zymosan. Methods: Biopsies from patients with chronic hyperplastic candidosis (n = 5), leukoplakia (n = 5), and healthy mucosa (n = 5) were immunohistochemically stained with antibodies to the TLRs (TLR1 to TLR9) to distinguish and compare the staining patterns of the epithelial layer in the three categories of tissues. Results: On analysis, the epithelium of all tissues was divided into three layers: basal, middle, and superficial. Two of the five chronic hyperplastic candidosis sections showed high numbers of hyphae compared to yeasts, which paralleled a decrease in the expression of TLR2 and an increase in the staining intensity of TLR4. Leukoplakia and healthy tissue sections demonstrated stronger immunostaining of TLRs, except TLR9 which showed weaker staining in some sections of the former, and in the basal layers of some sections of the latter. Discussion: This study supports the concept of negative regulation of TLRs that are either ligand‐bound (e.g. in chronic hyperplastic candidosis), or not stimulated (in healthy tissue). It also augments the opinion that C. albicans, through its hyphae rather than blastospore, may utilize TLRs, i.e. TLR2, to evade the immune system of the host. Leukoplakia seems to be more immunologically alert, which reduces the chances of worsening the already‐diseased tissue.     

Syndecan‐1 immunohistochemical expression in gingival tissues of chronic periodontitis patients correlated with various putative factors

Kotsovilis S, Tseleni‐Balafouta S, Charonis A, Fourmousis I, Nikolidakis D, Vrotsos JA. Syndecan‐1 immunohistochemical expression in gingival tissues of chronic periodontitis patients correlated with various putative factors. J Periodont Res 2010; 45: 520–531. © 2010 John Wiley & Sons A/S Background and Objective: Limited information is available on the expression and distribution of syndecan‐1 within human gingival tissues/cells and on putative factors that might affect its expression. Therefore, the objective of the present study was to determine immunohistochemically the expression and distribution of syndecan‐1 in the gingival tissues of patients with chronic periodontitis and to examine the correlation of syndecan‐1 expression with various putative factors (environmental, patient/systemic and local factors). Material and Methods: Gingival specimens were surgically excised from the area of the junctional/pocket epithelium (study group 1, including 30 chronic periodontitis patients) or the gingival oral epithelium (study group 2, comprising another 30 chronic periodontitis patients), adjacent to teeth with poor prognosis. Standard two‐step immunohistochemistry and semi‐quantitative evaluation of immunohistochemical staining were used to determine syndecan‐1 expression. Statistical analyses on the impact of various putative factors were performed. Results: In the junctional/pocket epithelium or the oral epithelium, syndecan‐1 expression was weak to moderate in the suprabasal and basal epithelial cells and absent to weak in the internal basal lamina, external basal lamina and gingival connective tissue matrix. Syndecan‐1 expression in the junctional/pocket epithelium was statistically significantly stronger than in the oral epithelium in inflammatory cells within the underlying gingival connective tissue (primarily plasma cells and lymphocytes) and in scattered fibroblast‐like cells. Conclusions: Syndecan‐1 expression in the junctional/pocket epithelium or the oral epithelium can exhibit a significant positive correlation with the severity/degree of histologically evaluated local gingival inflammation, but in general is not significantly correlated with age, smoking, full‐mouth and local clinical (probing pocket depth and clinical attachment level) and radiographical parameters (radiographical bone loss) of periodontal status.     

Expression of transient receptor potential vanilloid receptor 1 and toll-like receptor 4 in aggressive periodontitis and in chronic periodontitis

Öztürk A, Y?ld?z L. Expression of transient receptor potential vanilloid receptor 1 and toll‐like receptor‐4 in aggressive periodontitis and in chronic periodontitis. J Periodont Res 2011; 46: 475–482. © 2011 John Wiley & Sons A/S Background and Objective: The objective of the present study was to evaluate the expression and the distribution of the transient receptor potential vanilloid receptor 1 (TRPV1) and of toll‐like receptor 4 (TLR4) in tissue samples from patients with periodontal disease (aggressive periodontitis and chronic periodontitis) and from healthy controls. Material and Methods: Ten tissue samples from each disease group (aggressive periodontitis and chronic periodontitis) and from healthy subjects were obtained during routine oral surgical procedures. Subgingival specimens were collected from sites with advanced loss of support (probing depth > 5 mm) and specimens from the corresponding healthy controls were obtained during tooth extraction for orthodontic reasons or following surgical extraction of an impacted third molar. The distribution of TRPV1 and TLR4 receptors in human gingival tissue was studied by immunohistochemistry. Results: Both TLR4 and TRPV1 were detected in gingival tissues from healthy subjects, and from patients with chronic periodontitis and aggressive periodontitis, particularly in gingival keratinocytes, fibroblasts, inflammatory cells and the endothelial lining of capillaries in connective tissues. Histologic examination of the samples from healthy controls disclosed that clinically healthy gingiva does not correspond to histologically healthy gingiva. Subsequently, these samples were redesignated as gingivitis samples. TRPV1 was down‐regulated in all cell types in samples obtained from patients with chronic periodontitis compared to samples obtained from patients with gingivitis, whereas TLR4 was down‐regulated only in the epithelium and in gingival fibroblasts. In contrast, the levels of these markers in patients with aggressive periodontitis were similar to those in healthy patients. Conclusion: Local expression of TRPV1 and TLR4 in gingival tissues may contribute to both physiological and pathological processes in the periodontium. Our data suggest that TRPV1 and TLR4 may play a role specifically in the pathophysiology of chronic periodontitis.     

Immunohistochemical localization of CD1a and S100 in gingival tissues of healthy and chronic periodontitis subjects

Oral Diseases (2012) 18, 778-785 Objective: The aim of this study was to evaluate the presence and distribution of CD1a and S100 protein markers in states of gingival health and chronic periodontitis in human subjects. Materials and Methods: Gingival tissue samples were derived from 10 healthy and 10 chronic periodontitis-affected human subjects. The presence and distribution of CD1a and S100 protein was assessed using immunohistochemistry, and the cell types involved in their expression was determined. Results: The presence and distribution of CD1a was confined only to the gingival epithelium, whereas S100 was seen in the epithelium and connective tissue. However, increased expression of both CD1a and S100 protein was seen in periodontitis-affected gingival tissues compared with healthy gingiva. Immunohistochemistry demonstrated that CD1a- and S100-positive cells in the epithelium are Langerhans cells (LCs) and S100 positive cells in the connective tissue are dendritic cells (DCs). Conclusion: Our findings suggest the transition of CD1a-positive LCs to S100-positive DCs from epithelium to connective tissue in response to an antigenic challenge. Demonstration of increased number of S100-positive DCs in the gingival connective tissue in chronic periodontitis possibly suggests their involvement in bone resorption in addition to their antigen presentation property.     

Antimicrobial peptide hCAP‐18/LL‐37 protein and mRNA expressions in different periodontal diseases

Oral Diseases (2010) 17 , 60–67 Objective: To investigate the levels of antimicrobial peptide hCAP‐18/LL‐37 protein and mRNA expression in gingival tissues with different periodontal disease. Materials and methods: Ten patients with generalized aggressive periodontitis, 10 patients with chronic periodontitis, and 10 healthy controls were included in this study. Periodontal parameters including probing depth, clinical attachment level, plaque index, and papilla bleeding index were assessed in study subjects. hCAP‐18/LL‐37 mRNA analysis by RT‐PCR and immunohistochemistry were performed in 19 samples provided enough RNA in terms of concentration and integrity. Results: This study demonstrated that hCAP‐18/LL‐37 was a product of neutrophils. Tissue samples of chronic periodontitis patients had significantly higher immunostaining of hCAP‐18/LL‐37 on neutrophils infiltrating in both epithelium and connective tissue compared with controls. hCAP‐18/LL‐37 mRNA expression levels in tissue samples of chronic periodontitis patients seemed to be upregulated compared with controls. While two generalized aggressive periodontitis patients showed downregulated hCAP‐18/LL‐37 mRNA expression levels, one generalized aggressive periodontitis patient showed slightly increased hCAP‐18/LL‐37 mRNA level compared with controls. Conclusions: hCAP‐18/LL‐37 has an important role in innate response during periodontal inflammation. Local deficiency in hCAP‐18/LL‐37 might be a confounding effect in the pathogenesis of generalized aggressive periodontitis.     

Hepatocyte growth factor (HGF) system in gingiva: HGF activator expression by gingival epithelial cells

Hepatocyte growth factor (HGF) acts as a mitogen, motogen, morphogen, and anti-apoptotic factor for various kinds of epithelial cells. We previously showed that periodontal ligament and gingival fibroblasts secreted an HGF-like chemoattractant for a gingival epithelial cell line and found that the HGF content of gingival crevicular fluid was well correlated with clinical parameters and interleukin-1beta level. Since HGF is secreted as an inactive form (proHGF), and converted to an active form by serine proteases such as HGF activator (HGFA), extracellular processing of proHGF is presumed to be critical in the regulation of HGF activity. To examine the role of the HGF system in epithelial invasion followed by loss of connective tissue attachment in periodontitis, mRNA expression of HGF, its receptor (c-met) and HGFA in gingival tissues was monitored. Ten gingival biopsies were obtained, and epithelium and connective tissues were separated by enzymatic digestion. The gene expression of HGF and keratinocyte growth factor (KGF) in gingival connective tissue, and c-met, HGFA and KGF receptor (KGFR) in gingival epithelial tissues was monitored using RT-PCR. Furthermore, HGFA protein in the conditioned medium of cultured primary gingival epithelial cells was examined using Western blotting. All the connective tissue samples expressed KGF, and 8 out of 10 samples expressed HGF. All the epithelial samples expressed KGFR and c-met, whereas 5 out of 10 samples expressed HGFA. Protein expression of HGFA by cultured primary gingival epithelial cells was also confirmed. In terms of local production and activation of HGF in gingival tissue, these results suggest that synergistic expression of HGF in connective tissue and HGFA expression in epithelium may contribute to disease progression in periodontitis.     

Differential expression of Toll-like receptors in chronic hyperplastic candidosis

Introduction:  The first step in the host defense against oral candidosis is the recognition of Candida albicans through a set of germ-encoded pathogen recognition receptors, e.g. Toll-like receptors (TLRs). In man, 10 types of such receptors have been identified so far, of which only TLR2, TLR4, and TLR6 have been linked to mediating candidal ligands, e.g. zymosan.
Methods:  Biopsies from patients with chronic hyperplastic candidosis ( n  = 5), leukoplakia ( n  = 5), and healthy mucosa ( n  = 5) were immunohistochemically stained with antibodies to the TLRs (TLR1 to TLR9) to distinguish and compare the staining patterns of the epithelial layer in the three categories of tissues.
Results:  On analysis, the epithelium of all tissues was divided into three layers: basal, middle, and superficial. Two of the five chronic hyperplastic candidosis sections showed high numbers of hyphae compared to yeasts, which paralleled a decrease in the expression of TLR2 and an increase in the staining intensity of TLR4. Leukoplakia and healthy tissue sections demonstrated stronger immunostaining of TLRs, except TLR9 which showed weaker staining in some sections of the former, and in the basal layers of some sections of the latter.
Discussion:  This study supports the concept of negative regulation of TLRs that are either ligand-bound (e.g. in chronic hyperplastic candidosis), or not stimulated (in healthy tissue). It also augments the opinion that C. albicans , through its hyphae rather than blastospore, may utilize TLRs, i.e. TLR2, to evade the immune system of the host. Leukoplakia seems to be more immunologically alert, which reduces the chances of worsening the already-diseased tissue.     

纤维调节素在大鼠和人类牙龈组织中的差异性表达

目的:研究纤维调节素在大鼠和人类健康和牙龈组织中的分布与表达。方法:用免疫组织化学的方法研究纤维调节素在大鼠牙龈及人类健康和炎性牙龈组织中的分布。与之比较,I型胶原纤维在鼠磨牙牙龈组织中的分布也做了研究。结果:在大鼠牙龈中,纤维调节素分布在牙龈上皮基底上层,在腭侧牙龈结缔组织有强表达。I型胶原纤维也有类似的分布。比较而言,纤维调节素在颊侧牙龈结缔组织表达较弱。对于人类,纤维调节素在炎性牙龈结缔组织中表达明显,然而在健康牙龈结缔组织中表达较弱。结论:与Ⅰ型胶原纤维关系密切的纤维调节素在颊侧和腭侧牙龈中有差异性表达,并且在人类炎性牙龈组织中表达增强。     

Toll‐like receptors 2 and 5 in human gingival epithelial cells co‐operate with T‐cell cytokine interleukin‐17

Background/aim: Periodontitis begins as the result of perturbation of the gingival epithelial cells caused by subgingival bacteria interacting with the epithelial cells via pattern recognition receptors. Toll‐like receptors (TLRs) have been shown to play an important role in the recognition of periodontal pathogens so we have studied the interaction of TLR ligands with TLR2 and TLR5 for cytokine production in the cultures of gingival epithelial cells. Methods: Immunohistochemistry was used for the localization of TLR2 and TLR5 in tissue specimens. Enzyme‐linked immunosorbent assays were performed to detect the levels of interleukin‐1β (IL‐1β) and tumor necrosis factor‐α (TNF‐α), released from gingival epithelial cell cultures following stimulation with TLR ligand alone or in combination with IL‐17. Results: Both TLR2 and TLR5 were increased in periodontitis (2128 ± 159 vs. 449 ± 59 and 2456 ± 297 vs. 679 ± 103, respectively, P < 0.001) including gingival epithelial cells that stained strongly. Cultured gingival epithelial cells stimulated with their respective ligands (HKLM, a TLR2 ligand that is also found in Porphyromonas gingivalis, and flagellin, a TLR5 ligand that is also found in Treponema denticola) produced both IL‐1β and TNF‐α. To mimic T‐cell help, IL‐17 was added. This further greatly enhanced TLR ligand‐induced IL‐1β (P < 0.001) and TNF‐α (P < 0.01) production. Conclusions: These findings show how pathogen‐associated molecular patterns, shared by many different periodontopathogenic bacteria, stimulate the resident gingival epithelial cells to inflammatory responses in a TLR‐dependent manner. This stimulation may be particularly strong in periodontitis and when T helper type 17 cells provide T‐cell help in intercellular cooperation.     

老年牙周炎牙龈组织Fas/Apo-1(CD95)抗原表达的免疫组织化学研究

目的 :观察慢性老年牙周炎牙龈组织中Fas/Apo - 1(CD95 )抗原表达和分布情况。方法 :采用免疫组织化学染色方法对 10例慢性老年牙周炎患者、10例慢性成人牙周炎患者、10例青少年牙周炎患者和 10例健康老年人牙龈组织中Fas/Apo - 1(CD95 )抗原阳性表达和分布情况进行了观察和比较。结果 :在慢性老年牙周炎组完整的牙龈鳞状上皮间桥、核周胞浆Fas/Apo - 1(CD95 )抗原阳性表达和结缔组织中淋巴细胞Fas/Apo - 1(CD95 )抗原阳性表达为 4组中最强 ;健康老年人组上皮角化层Fas/Apo - 1(CD95 )抗原阳性表达为 4组中最强 ;慢性成人牙周炎组和青少年牙周炎组结缔组织中淋巴细胞和吞噬细胞Fas/Apo - 1(CD95 )抗原阳性表达强于健康老年人组 ;慢性老年牙周炎组上皮细胞和上皮细胞的细胞间桥Fas/Apo - 1(CD95 )抗原阳性表达例数明显高于慢性成人牙周炎组和青少年牙周炎组 (P <0 .0 5 ) ;健康老年人组结缔组织中淋巴细胞和吞噬细胞Fas/Apo - 1(CD95 )抗原阳性表达例数明显低于其它 3组 (P <0 .0 5 )。结论 :由于感染性炎症和衰老的影响 ,①牙周炎患者和健康老年人牙龈组织中鳞状上皮凋亡细胞发生部位与身体其他器官鳞状上皮细胞凋亡发生部位不同 ;②在慢性老年牙周炎患者牙龈组织中上皮细胞和炎性细胞对细胞凋亡易感性     

老年牙周炎牙龈组织诱导型一氧化氮合酶分布研究

目的 :观察慢性老年牙周炎患者牙龈组织中诱导型一氧化氮合酶 (iNOS)分布。方法 :采用免疫组织化学方法对 10例慢性老年牙周炎患者、10例慢性成人牙周炎患者、10例青少年牙周炎患者和 10例健康老年人牙龈组织中诱导型一氧化氮合酶分布进行了检测并比较研究。结果 :(1)牙周炎时牙龈组织中诱导型一氧化氮合酶主要在鳞状上皮细胞胞浆核周区颗粒状阳性表达 ,毛细血管壁内皮细胞、老化的胶原纤维及上皮下基底膜共同形成了一种乳头状轮廓样阳性表达形态 ,结缔组织和肉芽组织中各类炎症细胞也显阳性表达 ;(2 )慢性老年牙周炎组血管壁内皮细胞、结缔组织内炎症细胞、上皮乳头阳性表达例数明显低于青少年牙周炎组和慢性成人牙周炎组 (P <0 .0 5 )。血管壁内皮细胞和胶原纤维阳性表达例数低于健康老年人组 (P <0 .0 5 )。结论 :慢性老年牙周炎患者牙龈组织中诱导型一氧化氮合酶的表达明显降低 ,造成了局部一氧化氮(NO)合成减少 ,引起了局部牙龈组织免疫功能降低和免疫调节功能紊乱     

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.     

Immunohistochemical localization of very late activation integrins in healthy and diseased human gingiva

The β1-integrins (VLA family) are cellular adhesion molecules (CAM) that play a major role in cell-cell and cell-matrix interactions. The expression pattern of CAM was studied in 5 clinically normal volunteers with healthy gingiva and in 18 patients with clinically different stages of periodontitis. In healthy human gingiva α2. α3 and α6 integrin chains were found in a characteristic distribution, showing a broad continuous expression on the junctional and sulcular epithelium sites. The expression of these integrins was demonstrated primarily on the basal cell layers and in some cells of the stratum spinosum. Inflammatory stages of periodontitis revealed further upregulation of α2, α3 and α6 integrins into the junctional and sulcular epithelial cells, which correlated with the stage of the periodontitis and the extent of the cellular infiltration. α4 and α6 were found to be the predominant β1 integrin chains on inflammatory cells. The amount of δ4 and ş6 positive infiltrative cells increased with the number of inflammatory cells. VCAM-1. the corresponding cell-cell ligand of VLA-4 (α4) was present on the majority of subepithelial vessels in all stages of gingivitis and periodontitis. The α5 subunit was expressed on both endothelium and gingival connective tissue cells. Samples from advanced periodontitis cases showed a higher number of a5 positive mononuclear cells. In comparison to normal epidermis, a human gingival epithelial cells express higher levels of integrins. This expression is further upregulated in advanced stages of periodontitis, indicating changes of the β1 integrin organization.     

Toll like receptor 4 and membrane-bound CD14 expressions in gingivitis, periodontitis and CsA-induced gingival overgrowth

Objective

Immune cell recognition of lipopolysaccharides via CD14 and Toll-like receptor 4 (TLR4) complexes plays a crucial role in linking innate and adaptive immune responses. This study was aimed to investigate the expression of TLR4 and membrane-bound CD14 (mCD14) in the gingival tissues of patients with gingivitis, periodontitis and CsA-induced gingival overgrowth.

Design

Gingival tissues were obtained from 10 renal transplant patients receiving cyclosporine-A (CsA) and having gingival overgrowth (GO), 10 patients with chronic periodontitis, 10 generalized aggressive periodontitis, 10 gingivitis and 10 healthy subjects. Immunohistochemistry was performed in order to determine the localization of TLR4 and mCD14 in tissue specimens.

Results

TLR4 and mCD14 expressions were detected in all tissues including healthy gingival biopsies. TLR4 and mCD14 positive cells were predominantly confined to the epithelium–connective tissue interface area, and were highly expressed in the basal cell layer of patients with CsA GO and chronic periodontitis, compared to healthy group (P < 0.05).

Conclusion

The present study suggests that TLR4 and mCD14 protein expressions may be interrelated and appear to be associated with periodontal disease. CsA usage seemed not to affect TLR4 and mCD14 expressions in CsA induced GO tissues.     

Increased Expression of Interleukin (IL)‐35 and IL‐17, But Not IL‐27, in Gingival Tissues With Chronic Periodontitis

Background: Interleukin (IL)‐35 plays an important role in immune regulation through the suppression of effector T‐cell populations, including T‐helper 17 (Th17) cells. Although Th17 cells and IL‐17 are involved in the pathogenesis of periodontitis, the level of IL‐35 in inflamed periodontal tissues is unclear. Here, IL‐35, IL‐17, and IL‐27 production/expression in gingival crevicular fluid (GCF) and human gingival tissue were investigated. Methods: GCF samples were collected from buccal (mesial, center, and distal) sites of teeth from patients with chronic periodontitis (CP) and healthy controls and were analyzed by enzyme‐linked immunosorbent assay for IL‐35 (periodontitis, n = 36; healthy, n = 30) and IL‐17 (periodontitis, n = 16; healthy, n = 13). Gingival tissue, including sulcus/pocket epithelium and underlying connective tissue, was collected from an additional 10 healthy participants and 10 patients with CP and were analyzed by quantitative polymerase chain reaction (qPCR) for Epstein Barr virus‐induced gene 3 (EBI3), IL12A, and IL17A. IL27p28 was also tested by qPCR. Results: IL‐35 and IL‐17 were significantly higher in GCF from patients with periodontitis than healthy participants (P <0.01, P <0.05, respectively). In both healthy participants and those with periodontitis, positive correlations were found among IL‐35 and probing depth and clinical attachment level (CAL) as well as between IL‐17 and CAL. EBI3, IL12A (components of IL‐35), and IL17A messenger RNA expression levels were significantly higher in inflamed gingival tissue than in healthy control tissues (P <0.05). IL27p28 was not detected in any sample, suggesting that IL‐27 is not produced in large quantities in periodontal tissue. Conclusion: IL‐35 and IL‐17, but not IL‐27, may play important roles in the pathogenesis of periodontitis.