Serum interleukin-6 level is a useful marker in evaluating therapeutic effects of levamisole and Chinese medicinal herbs on patients with oral lichen planus

Background: Oral lichen planus (OLP) is a T cell‐mediated inflammatory disease. Interleukin‐6 (IL‐6) is a pro‐inflammatory cytokine that has effects on cellular and humoral immunities. Previous studies have shown that keratinocytes and tissue‐infiltrating mononuclear cells from OLP lesions can secrete IL‐6. In some OLP patients, the high serum IL‐6 levels are reduced after treatment, suggesting that IL‐6 may be a useful marker in evaluating therapeutic effects and in monitoring the disease status of OLP. Methods: In this study, we used a solid phase, two‐site sequential chemiluminescent immunometric assay to determine the baseline serum levels of IL‐6 in a group of 180 patients with erosive OLP (EOLP), nonerosive OLP (NEOLP), erythema multiforme (EM), traumatic ulcers (TU), oral submucous fibrosis (OSF), pemphigus vulgaris (PV), or Sjögren's syndrome (SS), and in 77 normal control subjects. Some OLP patients were treated with levamisole plus Chinese medicinal herbs or levamisole only for 0.5–5.5 months and their serum IL‐6 levels were measured after treatment. Results: We found that approximately 99% of the normal control subjects and the patients with EM, TU, or OSF had a normal serum IL‐6 level less than 5.0 pg/ml. However, 15% (22/149) OLP patients, 15% (20/136) EOLP patients, 20% (5/25) major type EOLP patients, 14% (15/111) minor type EOLP patients, 15% (2/13) NEOLP patients, 14% (1/7) EM patients, 43% (3/7) PV patients, and 100% (6/6) SS patients had a serum IL‐6 level greater than 5.0 pg/ml. The mean serum IL‐6 level in patients with OLP (3.4 ± 3.1 pg/ml, P < 0.001), EOLP (3.4 ± 3.2 pg/ml, P < 0.001), major type EOLP (4.9 ± 3.5 pg/ml, P < 0.001), minor type EOLP (3.0 ± 3.0 pg/ml, P < 0.01), or NEOLP (4.2 ± 1.5 pg/ml, P < 0.001) was significantly higher than that in normal control subjects (2.0 ± 1.5 pg/ml). A significant difference in the mean serum IL‐6 level was also found between major type and minor type EOLP patients (P < 0.01). The mean reduction of serum IL‐6 level in OLP patients treated with levamisole plus Chinese medicinal herbs was significantly higher (7.4 ± 4.7 pg/ml) than that in OLP patients treated with levamisole only (3.8 ± 2.3 pg/ml, P < 0.05), suggesting that the combination therapy was superior to levamisole only. Conclusion: We conclude that levamisole and levamisole plus Chinese medicinal herbs can modulate the serum IL‐6 level in OLP patients. IL‐6 may be a useful marker in evaluating therapeutic effects and in monitoring the disease status of OLP.     

Levamisole can modulate the serum tumor necrosis factor-α level in patients with recurrent aphthous ulcerations

BACKGROUND: Recurrent aphthous ulcerations (RAU) are common oral inflammatory lesions. Tumor necrosis factor (TNF)-alpha is an important inflammatory mediator and a critical cytokine for adequate host defense. Our previous studies have shown that 14-43% and 59-63% of patients in the ulcerative stage of major, minor or herpetiform RAU have significantly higher than normal serum levels of interleukin (IL)-6 and IL-8, respectively. In this study, we examined whether RAU patients in the ulcerative stage had a significantly higher than normal serum level of TNF-alpha and assessed whether treatment with levamisole can modulate serum TNF-alpha levels in RAU patients. METHODS: This study used a solid phase, two-site sequential chemiluminescent immunometric assay to determine the baseline serum levels of TNF-alpha in 146 patients with RAU, nine patients with traumatic ulcers (TU), and 54 normal control subjects. Fifty-five RAU patients with serum TNF-alpha levels higher than 5.0 pg/ml were treated with levamisole for 0.5-4 months and their serum TNF-alpha levels were measured after treatment. RESULTS: We found that 29% (42 of 146) RAU patients as well as 39% (24 of 61) major type, 20% (14 of 69) minor type, and 25% (four of 16) herpetiform type RAU patients had a serum level of TNF-alpha greater than the upper normal limit of 7.4 pg/ml. The mean serum level of TNF-alpha in patients with RAU (9.1 +/- 1.0 pg/ml, P < 0.001), major type RAU (11.6 +/- 1.9 pg/ml, P < 0.001), minor type RAU (6.9 +/- 0.9 pg/ml, P < 0.005), or herpetiform type RAU (9.6 +/- 2.7 pg/ml, P < 0.001) was higher than that (3.8 +/- 0.2 pg/ml) in normal control subjects. The mean serum TNF-alpha level was significantly higher in patients with major type RAU than in patients with minor type RAU (P < 0.05) and was significantly higher in major type RAU patients in the exacerbation stage than in the post-exacerbation stage (P < 0.05). In 55 RAU patients with serum TNF-alpha levels higher than 5.0 pg/ml, treatment with levamisole for a period of 0.5-4 months could significantly reduce the serum TNF-alpha level from 16.4 +/- 1.9 to 5.8 +/- 0.6 pg/ml (P < 0.001). CONCLUSIONS: We conclude that a significantly higher than normal serum level of TNF-alpha can be detected in 20-39% of patients in the ulcerative stage of major, minor or herpetiform RAU. The serum TNF-alpha level may be associated with the severity and the stage of RAU. Levamisole can modulate serum TNF-alpha levels in RAU patients.     

Levamisole and/or Chinese medicinal herbs can modulate the serum level of squamous cell carcinoma associated antigen in patients with erosive oral lichen planus

Abstract: The serum levels of squamous cell carcinoma associated antigen (SCCA) were determined by a microparticle enzyme immunoassay in a group of patients with stage I oral squamous cell carcinoma (OSCC), major or minor type erosive oral lichen planus (EOLP), recurrent aphthous stomatitis (RAS), Behçet’s disease (BD), oral leukoplakia (OL), or oral submucous fibrosis (OSF), and in normal control subjects. About 97% of the normal control subjects and the patients with minor type EOLP, RAS, BD, OL or OSF had a serum level of SCCA within the normal limit of 1.2 ng/ml. However, 6 of the 12 (50%) patients with stage I OSCC and 14 of the 31 (45.2%) patients with major type EOLP had a serum level of SCCA greater than 1.2 ng/ml. The mean serum level of SCCA in stage I OSCC patients (1.38±1.16 ng/ml) or in major type EOLP patients (1.32±1.23 ng/ml) was significantly higher than that in normal control subjects (P<0.001) and that in the patients with minor type EOLP (P<0.001), RAS (P<0.001), BD (P<0.05), OL (P<0.05), or OSF (P<0.05). Either major or minor type EOLP patients could obtain a significant mean reduction of the serum SCCA level of 0.34–0.63 ng/ml after treatment with levamisole and/or Chinese medicinal herbs for 1–30 months. Combination therapy with levamisole plus Chinese medicinal herbs could achieve a shorter duration of treatment to get complete remission than the single therapy with either levamisole only or Chinese medicinal herbs only. We conclude that levamisole and/or Chinese medicinal herbs can modulate the serum SCCA level in EOLP patients. SCCA may be a useful marker in evaluating therapeutic effects and in monitoring the disease status of EOLP. For EOLP patients, the combination therapy is superior to the single therapy of levamisole or of Chinese medicinal herbs.     

Serum interleukin-8 level is a more sensitive marker than serum interleukin-6 level in monitoring the disease activity of recurrent aphthous ulcerations

BACKGROUND: Recurrent aphthous ulcerations (RAU) are common oral inflammatory lesions. Interleukin (IL)-8 is a pro-inflammatory cytokine of host response to injury and inflammation. Our recent study has found that measurement of serum IL-6 level can detect only 24% RAU patients with an abnormal serum level. In this study, we examined both the serum IL-6 and IL-8 levels in a group of RAU patients. The abilities of IL-6 and IL-8 to detect patients with an abnormal serum level were compared in order to find out whether IL-8 was a more sensitive serum marker than IL-6 in monitoring the disease activity of RAU. METHODS: In this study, we used a solid-phase, two-site sequential chemiluminescent immunometric assay to determine the baseline serum levels of IL-6 and IL-8 in 146 patients with RAU, 9 patients with traumatic ulcers (TU), and 54 normal control (NC) subjects. Eighty-two RAU patients, with the serum IL-6 or IL-8 levels higher than the upper limit of normal serum concentration, were treated with levamisole for 0.5-3.5 months, and their serum IL-6 and IL-8 levels were measured after treatment. RESULTS: We found that 25% (37/146) RAU patients, as well as 33% (20/61) major-type, 19% (13/69) minor-type, and 25% (4/16) herpetiform-type RAU patients, had a serum level of IL-6 greater than the upper normal limit of 4.7 pg/ml. In contrast, 60% (87/146) RAU patients, as well as 59% (36/61) major-type, 59% (41/69) minor-type, and 63% (10/16) herpetiform-type RAU patients, had a serum level of IL-8 greater than the upper normal limit of 8.7 pg/ml. In 82 RAU patients with the serum IL-6 or IL-8 levels higher than the upper limit of normal serum concentration, treatment with levamisole for a period of 0.5-3.5 months could significantly reduce the serum IL-6 level from 12.0 +/- 1.6 to 3.0 +/- 0.5 pg/ml (P < 0.001), and could significantly lower the serum IL-8 level from 70.9 +/- 11.2 to 13.8 +/- 3.1 pg/ml (P < 0.001). CONCLUSIONS: Because measurement of serum IL-8 level can detect 60% RAU patients with an abnormal serum level, while measurement of serum IL-6 level can detect only 25% RAU patients with an abnormal serum level, we conclude that serum IL-8 level is a more sensitive marker than serum IL-6 level in monitoring the disease activity of RAU. Levamisole can modulate both the serum IL-6 and IL-8 levels in RAU patients. IL-8, like IL-6, is also a useful serum marker in evaluating therapeutic effects of levamisole on RAU patients.     

Treatment with levamisole and colchicine can result in a significant reduction of IL-6, IL-8 or TNF-α level in patients with mucocutaneous type of Behcet's disease

Background:  Mucocutaneous type of Behcet's disease (MCBD) is a multisystemic inflammatory disease with oral and genital ulcers with or without skin lesions.
Methods:  A solid phase, two-site sequential chemiluminescent immunometric assay was used to measure serum levels of interleukin (IL)-6, IL-8 and tumour necrosis factor (TNF)-α in 54 normal control subjects and in 64 MCBD patients before and after treatment with levamisole plus colchicine.
Results:  We found that 67%, 83% or 67% of MCBD patients had a serum IL-6, IL-8 or TNF-α level greater than the upper normal limit of 4.7, 8.7 or 7.4 pg/ml, respectively. The mean serum level of IL-6 (9.9 ± 2.4 pg/ml, P  < 0.005), IL-8 (107.5 ± 21.4 pg/ml, P  < 0.001) or TNF-α (22.5 ± 4.1 pg/ml, P  < 0.001) in 64 MCBD patients was significantly higher than that (2.1 ± 0.2, 5.7 ± 0.2 or 3.8 ± 0.2 pg/ml for IL-6, IL-8 or TNF-α level, respectively) in normal control subjects. In 43 MCBD patients with all the serum IL-6, IL-8 and TNF-α levels higher than their upper normal limits, treatment with levamisole plus colchicine for a period of 0.5–11.5 (mean, 3.2 ± 2.4) months could significantly reduce the mean serum IL-6, IL-8 and TNF-α levels from 9.0 ± 1.7 to 1.6 ± 0.2 pg/ml ( P  < 0.001), 134.6 ± 28.2–6.0 ± 0.4 pg/ml ( P  < 0.001) and 25.7 ± 5.6–3.5 ± 0.4 pg/ml ( P  < 0.001), respectively.
Conclusions:  Treatment with levamisole and colchicine can result in a significant reduction of serum IL-6, IL-8 or TNF-α level in MCBD patients.     

Serum cytokine and T regulatory cell levels in patients with burning mouth syndrome

Background: Burning mouth syndrome is a disorder usually associated with an unexplained, prolonged sensation of burning inside the oral cavity. Although the etiology is unknown, neural and psychologic factors and cytokines may be implicated in the pathogenesis of burning mouth syndrome. The aim of this study was to investigate the relationship between serum cytokine and T regulatory cell levels in patients with burning mouth syndrome with regard to depression and anxiety. Methods: Thirty patients with burning mouth syndrome and 30 matched controls participated in the study. Serum cytokine levels were measured with cytometric bead array and T regulatory cells were defined as CD4⁺CD25⁺Foxp‐3⁺ cells by flow cytometry. The level of anxiety and depression were analyzed by means of the Speilberger State‐Trait Anxiety Inventory and Zung Self‐Rating Depression Scale. Visual analogue scale was used in the quantification of burning levels of patients. Results: Serum IL‐2 and TNF‐α levels were significantly decreased in patients with burning mouth syndrome compared with controls [mean 16.79 ± 8.70 vs. 37.73 ± 41.05 pg / ml (P < 0.05) and mean 39.09 ± 29.40 vs. 70.83 ± 42.44 pg / ml (P < 0.01) respectively]. Conclusions: IL‐2 and TNF‐α might play a role in burning mouth syndrome. Burning mouth syndrome may occur as a sign of predisposition to autoimmunity. Presence of low levels of CD28⁺ supports the provision that BMS might be a pre‐otoimmune disease.     

复发性阿弗他溃疡患者25羟维生素D水平的研究

目的 比较轻型复发性阿弗他溃疡患者与健康人血清25羟维生素D的水平,分析25羟维生素D与复发性阿弗他溃疡发病机制间可能存在的关系.方法 选取48名确诊为轻型复发性阿弗他溃疡的患者和50名健康人为研究对象.分别在患者发病期(A1组)和无症状期(A2组)抽取5ml空腹静脉血,以健康人空腹静脉血为对照组.采用酶联免疫法检测血清25羟维生素D水平.结果 A1组25羟维生素D水平最低(14.73±5.18)ng/ml,显著低于A2组(17.36±5.96)ng/ml差异有统计学意义(P<0.05).对照组25羟维生素D水平最高(19.87±5.94)ng/ml,与A1、A2组差异均有统计学意义(P<0.05).结论 维生素D可能在复发性阿弗他溃疡的发病机制中具有一定的作用.     

The Effects of Initial Periodontal Therapy on the Serum Receptor Activator of Nuclear Factor‐κβ Ligand/Osteoprotegerin System in Patients With Type 2 Diabetes Mellitus and Periodontitis

Background: The aim of the present study is to evaluate the serum receptor activator of nuclear factor‐κβ ligand (RANKL)/osteoprotegerin (OPG) system in patients with chronic periodontitis (CP) and type 2 diabetes mellitus (T2DM) and its changes after periodontal intervention. Methods: Thirty‐five patients with CP + T2DM, 35 systemically healthy patients with CP, and 35 healthy controls were enrolled, and serum levels of RANKL and OPG were measured at baseline. Then the CP + T2DM group was divided into a well‐controlled subgroup and a poorly controlled subgroup according to their hemoglobin A1c (HbA1c), and initial periodontal therapy was performed. After 3 months, patients in both subgroups were recalled, and serum RANKL and OPG levels were tested again and compared with the baseline. Results: At baseline, serum levels of OPG in the T2DM + CP group were much lower than in the CP group and healthy controls (197.41 ± 57.05 pg/mL versus 232.60 ± 70.85 pg/mL [CP group] or 244.96 ± 85.13 pg/mL [healthy controls], P <0.05), whereas their RANKL levels were much higher than in the other two groups (324.35 ± 87.62 pg/mL versus 284.52 ± 90.35 pg/mL [CP group] or 163.01 ± 45.24 pg/mL [healthy control], P <0.05), as was the RANKL/OPG (R/O) ratio (1.68 ± 0.33 versus 1.26 ± 0.35 [CP group] or 0.72 ± 0.25 [healthy control], P <0.001). Serum levels of OPG in both disease groups had significant negative correlations with HbA1C, and serum levels of RANKL in all participants had significant positive correlations with periodontal parameters. After periodontal intervention, both the well‐controlled and poorly controlled subgroups exhibited significant increases in OPG and decreases in RANKL in serum, and the R/O ratio was also notably reduced. Additionally, the poorly controlled subgroup exhibited a greater reduction in HbA1c and a greater increase in OPG than the well‐controlled subgroup. Conclusions: The changing trend in the serum RANKL/OPG system in patients with T2DM + CP was similar to that seen in CP patients and may be even more pronounced. Periodontal intervention effectively improved glucose metabolism and changed the serum RANKL/OPG system regardless of whether patients’ HbA1c was well‐controlled or poorly controlled over the 3‐month observation period.     

Retrospective study of two biochemical markers for the risk assessment of oral bisphosphonate‐related osteonecrosis of the jaws: can they be utilized as risk markers?

Purpose: The purpose of this retrospective study was to examine the possibility of utilizing serum C‐terminal telopeptide cross‐link of type I collagen (s‐CTX) and serum osteocalcin (s‐OC) as risk markers for oral bisphosphonate‐related osteonecrosis of the jaws (BRONJ). Patients and methods: The s‐CTX values and the s‐OC values were measured from 23 patients (one male, 22 females) diagnosed with BRONJ using clinical and radiographic examinations. The two biochemical markers were evaluated during a regular checkup for osteoporosis management. For the control group of s‐CTX study, s‐CTX values were obtained from 61 independently recruited postmenopausal women who have been on bisphosphonate therapy for >6 months. The s‐CTX values of the ONJ group and the control group were compared. Because of retrospective nature of this study, the control group for s‐OC study could not be established. A single sample t‐test was performed for the s‐OC value from the ONJ group. Result: Twenty‐three ONJ patients had taken alendronate for osteoporosis treatment, and the s‐CTX testing results were low levels of 10–192 pg/ml (mean: 93.2±49.4 pg/ml). Mean of s‐CTX of the control (n=61) was 125±85.7 pg/ml. The duration of BP therapy ranged between 1 and 10 years (4.82±2.6). The s‐OC level was estimated between 0.2 and 5.4 ng/ml (1.91±1.51 ng/ml). The mean s‐CTX value of the control group was higher but without significance (P=0.12). The s‐OC values of the ONJ group were significantly lower than the lowest value of the reference range (P<0.001). Conclusion: As a result of the s‐CTX and s‐OC testings at the diagnosis of BRONJ, the values of the two markers were decreased. The decrease of the s‐OC values implies a problem during the bone‐formation process. Therefore, we can assume that in this patient group, invasive dental surgery contributes to an increase in the risk of BRONJ incidence. This result may imply that, during bisphosphonate therapy, simultaneous consideration of s‐CTX showing inhibition of bone resorption and s‐OC indicating the degree of bone formation might be a set of risk markers assessing risk prediction for BRONJ before invasive dental surgery. To cite this article:
Kwon Y‐D, Ohe J‐Y, Kim D‐Y, Chung D‐J, Park Y‐D. Retrospective study of two biochemical markers for the risk assessment of oral bisphosphonate‐related osteonecrosis of the jaws: can they be utilized as risk markers?
Clin. Oral Impl. Res. 22 , 2011; 100–105.
doi: 10.1111/j.1600‐0501.2010.01965.x     

Whole Salivary Interleukin‐6 and Matrix Metalloproteinase‐8 Levels in Patients With Chronic Periodontitis With and Without Prediabetes

Background: The cytokine profile in unstimulated whole saliva (UWS) of patients with prediabetes and chronic periodontitis (CP) remains uninvestigated. The aim of this study is to assess interleukin (IL)‐6 and matrix metalloproteinase (MMP)‐8 levels in UWS of patients with CP with and without prediabetes. Methods: Eighty‐eight males (aged 39 to 51 years) were divided into three groups: group 1: 28 patients with CP and prediabetes; group 2: 30 patients with CP and without prediabetes; and group 3: 30 controls. Fasting blood glucose (FBG) and hemoglobin A1c (HbA1c) levels, periodontal parameters (plaque index, bleeding on probing, probing depth, attachment loss, and marginal bone loss), and number of missing teeth were recorded. UWS samples were collected, and UWS flow rate (UWSFR) was measured. IL‐6 and MMP‐8 were measured in UWS using enzyme‐linked immunosorbent assay. P values <0.05 were considered statistically significant. Results: Mean FBG and HbA1c levels were significantly higher in group 1 (119.3 ± 3.1 mg/dL and 6.1% ± 0.2%) than group 2 (80.1 ± 3.5 mg/dL and 4.8% ± 0.5%; P <0.001) and group 3 (75.3 ± 2.2 mg/dL and 4.3% ± 0.2%; P <0.05). UWSFR was significantly higher in groups 2 (0.53 ± 0.1 mL/minute; P <0.05) and 3 (0.51 ± 0.1 mL/minute; P <0.01) than group 1 (0.33 ± 0.05 mL/minute). Periodontal parameters were worse in group 1 (P <0.05) and group 2 (P <0.05) than group 3. There was no difference in periodontal parameters, numbers of missing teeth, or salivary IL‐6 and MMP‐8 levels between patients in groups 1 and 2. Conclusion: Salivary IL‐6 and MMP‐8 levels are elevated in patients with CP with and without prediabetes.     

Interleukin‐2, interleukin‐6 and T regulatory cells in peripheral blood of patients with Behçet’s disease and recurrent aphthous ulcerations

J Oral Pathol Med (2012) 41 : 73–79 Background: One of the factors involved in the pathogenesis of Behçet disease (BD) and recurrent aphthous ulcerations (RAU) is a cell‐mediated immune response in which several cytokines (interleukin‐2, interleukin‐6) and T regulatory cell (T reg cell) population seem to play a major role. The aim of this study was to measured the interleukin‐2 (IL‐2), interleukin‐6 (IL‐6) levels and analysis of CD4⁺ CD25⁺ Foxp‐3⁺ Treg cells in peripheral blood from patients with BD and RAU. In addition; we also analysed peripheral blood from healthy subjects for comparison. Methods: Thirty patients (15 men and 15 women) with BD, 30 patients (12 men and 18 women) with RAU and 15 healthy control subjects (nine men and six women) participated in the study. Analysis of CD4⁺ CD25⁺ Foxp‐3⁺ Treg cells, IL‐2 and IL‐6 levels have been measured in flow cytometry. Results: No statistical differences were observed in the serum levels of IL‐2 and IL‐6 between BD and RAU patients, and healthy subjects. Although there were no statistical differences in the number of CD4⁺ CD25⁺ Foxp‐3⁺ cells between groups, there were statistically significant differences in the number of CD4⁺ CD25^bright Treg cells. CD4⁺ CD25^bright Treg cells were significantly increased in BD and RAU patients compared to healthy subjects. Statistical analysis revealed no difference according to the number of CD4⁺ CD25^bright cells between BD and RAU patients. Conclusions: These results indicate that CD4⁺ CD25^bright T regulatory cells may be contributing factor in the pathogenesis of BD and RAU.     

Proinflammatory and Anti‐Inflammatory Cytokines in Gingival Crevicular Fluid and Serum of Patients With Rheumatoid Arthritis and Patients With Chronic Periodontitis

Background: The aim of this study is to evaluate proinflammatory and anti‐inflammatory cytokine levels in gingival crevicular fluid (GCF) and serum of rheumatoid arthritis (RA) and chronic periodontitis (CP) patients to assess whether cytokine profiles distinguish patients with RA and patients with CP while using healthy patients as background controls. Methods: A total of 49 patients, 17 patients with RA (three males and 14 females; mean age: 47.82 ± 10.74 years), 16 patients with CP (10 males and six females; mean age: 44.00 ± 7.00 years), and 16 controls (eight males and eight females; mean age: 28.06 ± 6.18 years) were enrolled. Patients with RA were under the supervision of rheumatologists; 15 of the patients with RA were being treated with methotrexate–sulfasalazine combined therapy, and two of the patients were being treated with leflunomid therapy. Periodontal parameters (plaque index, gingival index, probing depth, and clinical attachment level) were recorded. Interleukin (IL)‐1β, IL‐4, IL‐10, and tumor necrosis factor‐α (TNF‐α) were determined in GCF and IL‐1β and IL‐10 in serum by enzyme‐linked immunosorbent assay. Results: There were significant differences found among RA, CP, and control groups for all periodontal parameters (P <0.05). The total amount and concentration of GCF IL‐1 β, IL‐4, IL‐10, and TNF‐α were similar in RA and CP patients (P >0.05). Although the total amount and concentration of serum IL‐10 was not significantly different among the groups (P >0.05), serum IL‐1β was significantly lower in the RA group compared to CP patients and controls and was higher in GCF of the RA group compared to the CP group. Conclusions: Although clinical periodontal disease parameters indicated more severe periodontal disease in CP compared to RA patients, immunologic evaluation did not reveal consistent results regarding proinflammatory and anti‐inflammatory cytokine levels. This might be a result of the use of non‐steroidal anti‐inflammatory drugs and rheumatoid agents by patients with RA.     

The influence of sex hormones on proinflammatory cytokines in gingiva of periodontally healthy premenopausal women

Markou E, Boura E, Tsalikis L, Deligianidis A, Konstantinidis A. The influence of sex hormones on proinflammatory cytokines in gingiva of periodontally healthy premenopausal women. J Periodont Res 2011; 46: 528–532. © 2011 John Wiley & Sons A/S Background and Objective: The aim of this work was to investigate any correlation between the fluctuation of levels of specific proinflammatory cytokines in gingival crevicular fluid and the fluctuation of sex hormones in peripheral blood at ovulation and progesterone peak. Material and Methods: Eighteen premenopausal women with normal and consistent menstrual cycles and healthy periodontium were included in this study. The exclusion criteria were as follows: (i) pregnancy; (ii) use of oral contraceptives; (iii) metabolic or systemic disease that might affect the periodontium; (iv) use of antimicrobial or nonsteroidal anti‐inflammatory drugs during the past 6 mo; and (v) smoking. The measurements were performed at two specific time points for each participant [(i) on the day of ovulation; and (ii) on the day of the progesterone peak) and included the following: (i) plaque index; (ii) bleeding on probing; and (iii) the gingival crevicular fluid levels of interleukin (IL)‐1β, IL‐6, IL‐8 and tumor necrosis factor‐α (TNF‐α). Results: During the menstrual cycle, plaque index values remained unchanged (0.71 ± 0.07 at ovulation; 0.73 ± 0.08 at progesterone peak; p > 0.05), as did bleeding on probing (0.35 ± 0.07 at ovulation; 0.41 ± 0.07 at progesterone peak; p > 0.05). At ovulation, mean gingival crevicular fluid levels were as follows: IL‐1β, 13.3 pg/sample; IL‐6, 5.9 pg/sample; IL‐8, 18.7 pg/sample; and TNF‐α, 25.9 pg/sample. The corresponding values at progesterone peak were as follows: 14.1, 10.1, 19.5 and 26.3 pg/sample. Only IL‐6 gingival crevicular fluid levels were significantly different between ovulation and progesterone peak (p < 0.05). This could reflect sensitivity to subclinical amounts of plaque and biofilm constituents. Conclusion: The subclinical increase of IL‐6 at progesterone peak is not accompanied by clinical changes in the periodontium.     

Evaluation of periodontal status and effectiveness of non-surgical treatment in patients with type 2 diabetes mellitus in Taiwan for a 1-year period

Background: The periodontal status and effects of non‐surgical periodontal treatment in patients with type 2 diabetes mellitus and periodontal disease are assessed. Methods: One‐hundred patients with type 2 diabetes (mean ± SD hemoglobin (Hb)A1c level: 7.3% ± 0.94%) and periodontal disease were recruited for this study. The group with moderate‐to‐severe periodontal disease included patients with >1 tooth with a probing depth (PD) ≥5 mm and >2 teeth with a clinical attachment loss (AL) ≥6mm, and the group with mild periodontal disease included patients with <1 affected tooth, and >2 affected with a clinical AL ≥6mm. Patients (28 patients in the mild group and 72 patients in the moderate‐to‐severe group) underwent non‐surgical periodontal treatments. We analyzed differences in serum concentrations of metabolic parameters (glycated hemoglobin and low‐density lipoprotein), inflammatory parameters (interleukin [IL]‐1β and C‐reactive protein [CRP]), and periodontal parameters between the two groups before treatment and at 3, 6, 9, and 12 months post‐therapy. Results: Seventy‐five patients with diabetes (21 patients in the mild group and 54 patients in the moderate‐to‐severe group) completed the study. Significant differences in the plaque index (PI), gingival index (GI), PD, and clinical AL at examination times were observed in the whole cohort (P <0.05). We observed significant differences in the PI, GI, and PD in the moderate‐to‐severe group (P <0.05), whereas there was only a significant difference in PD in the mild group (P <0.05) between baseline and 12 months post‐treatment. Both groups experienced improved glycemic control, but the difference was insignificant. CRP and IL‐1β levels were significantly different at examination times for the whole cohort (P <0.05). No significant positive association among metabolic and inflammatory parameters at 12 months post‐therapy were found. Conclusion: Non‐surgical periodontal treatment improved and maintained the periodontal health of patients with well‐controlled diabetes, but no significant reduction of metabolic parameters was observed over a 1‐year period.     

Multiplexed immunobead‐based assay for detection of oral cancer protein biomarkers in saliva

Objective: For clinical applications of biomarkers, there is a need for multiplex assays using high throughput platforms. The objective of this study was to determine the efficacy of Luminex Multianalyte Profiling (xMAP) technology for measurement of salivary proteins and to evaluate whether multiplex assays are as effective as single‐plex assays and enzyme‐linked immunosorbent assay (ELISA). Results: The average levels of interleukin‐8 (IL‐8) from the single‐plex assay were 3313.2 ± 3759.8 pg ml^?1 [oral squamous cell carcinoma (OSCC), n = 20] and 1061.7 ± 1978.8 pg ml^?1 (control, n = 20). The IL‐1β average levels from the single‐plex assay were 945.2 ± 1134.8 pg ml^?1 (OSCC, n = 20) and 314.2 ± 444.8 pg ml^?1 (control, n = 20). The average levels of IL‐8 from the multiplex assay were 2834.9 ± 3385.6 pg ml^?1 (OSCC, n = 20) and 947.3 ± 2036.8 pg ml^?1 (control, n = 20). The IL‐1β average levels from the multiplex assay were 1013.5 ± 1221.1 pg ml^?1 (OSCC, n = 20) and 376.3 ± 576.3 pg ml^?1 (control, n = 20). The correlation coefficient between Luminex and ELISA assay for IL‐8 (n = 19) and IL‐1β (n = 19) was 0.91 and 0.84, respectively. Conclusion: Luminex xMAP single‐plex and multiplex assays are as effective as ELISA assays for quantification of proteins in saliva. Both IL‐8 and IL‐1β were expressed at significantly higher levels in OSCC subjects than in the matched healthy control subjects.     

Platelet-activating factor levels of serum and gingival crevicular fluid in nonsmoking patients with periodontitis and/or coronary heart disease

The purpose of the present study was to investigate systemic and local levels of platelet-activating factor (PAF), a potent proinflammatory mediator implicated in cardiovascular pathophysiology in adult nonsmoking patients with periodontitis with or without coronary heart disease (CHD). Eighty-seven volunteers, 25 periodontitis patients, 19 periodontitis with CHD patients, 19 CHD patients, and 24 healthy controls were included, and periodontal conditions were assessed. Gingival crevicular fluid (GCF) and venous blood were collected, and PAF levels were measured by enzyme-linked immunosorbent assay. PAF levels in serum (303.3 ± 204 pg/ml) and in GCF (26.3 ± 6 pg/μl) of the periodontitis group with CHD, the periodontitis group (serum, 302.4 ± 241 pg/ml and GCF, 26.3 ± 8 pg/μl) and the CHD group (serum, 284.7 ± 192 pg/ml and GCF, 20.8 ± 6 pg/μl) were significantly higher than the healthy control group (serum, 65.4 ± 35 pg/ml and GCF, 7.7 ± 3 pg/μl; p < 0.05). In summary, the present study could demonstrate that in patients with periodontitis, the inflammatory mediator PAF is released into serum at least in the same range as for patients with coronary heart disease. However, no additive effects were seen when both conditions were present.     

Assessment of Interleukin‐6 Receptor Inhibition Therapy on Periodontal Condition in Patients With Rheumatoid Arthritis and Chronic Periodontitis

Background: Overproduction of interleukin (IL)‐6 may play a pathologic role in rheumatoid arthritis (RA) and chronic periodontitis (CP). The present study assesses IL‐6 receptor (IL‐6R) inhibition therapy on the periodontal condition of patients with RA and CP. Methods: The study participants were 28 patients with RA and CP during treatment with IL‐6R inhibitor, and 27 patients with RA and CP during treatment without IL‐6R inhibitor. Periodontal and rheumatologic parameters and serum levels of cytokine and inflammatory markers and immunoglobulin G against periodontopathic bacteria were examined after medication with IL‐6R inhibitor for 20.3 months on average (T1) and again 8 weeks later (T2). Results: No differences were observed between the groups in any parameter values at T1, except for serum IL‐6 levels. The anti–IL‐6R group showed a significantly greater decrease in gingival index, bleeding on probing (BOP), probing depth (PD), clinical attachment level (CAL), and serum levels of IL‐6 and matrix metalloproteinase (MMP)‐3 from T1 to T2 than the control group (P <0.05). A significant correlation was found between changes in serum anticyclic citrullinated peptide levels and those in PD and CAL in the anti–IL‐6R group (P <0.05), whereas both groups exhibited a significant association between changes in serum MMP‐3 levels and those in BOP (P <0.05). Conclusion: Changes in periodontal and serum parameter values were different between the patients with RA and CP during treatment with and without IL‐6R inhibitor.     

Immunomodulation by levamisole in patients with recurrent aphthous ulcers or oral lichen planus

The purpose of this study was to evaluate the effect of levamisole on the immune system of patients with recurrent aphthous ulcers (RAU) or oral lichen planus (OLP) in an open trial. Lymphocyte subsets, serum immunoglobulins, and circulating immune complexes (CIC) in patients with RAU or OLP and in normal control subjects were determined by an indirect immunofluorescence (ML) technique with monoclonal anti-lymphocyte antibodies, by single radial immunodiffusion, and by precipitation with 3% polyethylene glycol, respectively. In addition, the anti-nuclear antibodies (ANA) and anti-basal cell antibodies (anti-BCA) in sera were detected by an IIF technique. We found a significant improvement in clinical symptoms and normalization of the decreased CD4/CD8 ratio in RAU patients after levamisole treatment. Moreover, the decreased CD4/CD8 ratio, which persisted until the remission stage in the untreated RAU patients, reverted to normal m the active late stage in the levamisole-treated patients. This reversion of aberrant cellular immunity in an earlier stage of the ulcer cycle may explain why RAU patients experience symptom improvement after levamisole treatment. Although RAU patients treated with levamisole for 1 to 3 or 4 months still had higher than normal levels of CIC and serum immunoglobulins, the levels of their IgA and IgM returned to normal values after 4 months of levamisole treatment. The serum ANA detected m 6 patients with RAU and 3 patients with erosive OLP disappeared after 1-22 months of levamisole treatment. The disappearance of serum anti-BCA was also observed in 50%, of the anti-BCA-positive patients with erosive OLP after 3–13 months of levamisole treatment. These findings suggest that levamisole has modulating effects on both cell-mediated and humoral immunity in patients with RAU or OLP.     

Interleukin-4, a T-helper 2 cell cytokine, is associated with the remission of periodontal disease

Background and Objectives: Interleukin‐4 (IL‐4), secreted mainly by T‐helper 2 cells, is a key cytokine for the growth and proliferation of B lymphocytes. Previous studies have proved that IL‐4 has an anti‐inflammatory effect owing to its efficient inhibition of the production of proinflammatory cytokines such as tumour necrosis factor‐α (TNF‐α), IL‐1α, IL‐1β, IL‐6 and IL‐8 by monocytes/macrophages. The aim of the present study was to assess the relation between clinical parameters and concentrations of IL‐4 within gingival crevicular fluid from inflamed gingiva and periodontitis sites and, subsequently, after treatment of the periodontitis sites. Material and Methods: A total of 60 subjects were divided into three groups based on gingival index (GI), pocket probing depth and clinical attachment loss (CAL): healthy (group 1), gingivitis (group 2) and chronic periodontitis (group 3). A fourth group (group 4) consisted of 20 subjects from group 3, 6–8 weeks after treatment (i.e. scaling and root planing). Gingival crevicular fluid samples collected from each patient were quantified for IL‐4 using the enzymatic immunometric assay. Results: The highest mean concentration of IL‐4 was obtained for group 1 (99.39 ± 49.33 pg/mL) and the lowest mean concentration of IL‐4 was obtained for group 3 (15.78 ± 21.92 pg/mL). The mean IL‐4 concentrations for group 2 (64.34 ± 39.56 pg/mL) and group 4 (68.92 ± 42.85 pg/mL) were intermediate between the levels in healthy subjects and periodontitis subjects. Conclusion: The mean concentration of IL‐4 decreased from periodontal health to disease. Thus, we suggest that type 2 helper T cell cytokine, as represented by IL‐4, was associated with the remission or improvement of periodontal disease.     

Comparison of CCL28, interleukin‐8, interleukin‐1β and tumor necrosis factor‐alpha in subjects with gingivitis,chronic periodontitis and generalized aggressive periodontitis

Background and Objective: Cytokines produced by various cells are strong local mediators of inflammation. Mucosa‐associated epithelial chemokine (CCL28), interleukin‐8 (IL‐8), interleukin‐1beta (IL‐1β) and tumor necrosis factor‐alpha (TNF‐α) are major cytokines that play important roles in the periodontal inflammatory process. In this study we aimed to compare the levels of CCL28, IL‐8, IL‐1β and TNF‐α in the gingival crevicular fluid of both periodontally healthy subjects and in subjects diagnosed with gingivitis, chronic periodontitis and generalized aggressive periodontitis. Material and Methods: A total of 84 subjects participated in the study: 21 subjects had gingivitis, 21 subjects had chronic periodontitis, 21 subjects had generalized aggressive periodontitis and 21 were periodontally healthy. The levels of CCL28, IL‐8, IL‐1β and TNF‐α were analyzed using enzyme‐linked immune sorbent assay (ELISA). Results: The total levels of CCL28 and IL‐8 in the gingival crevicular fluid of the generalized aggressive periodontitis group (324.74 ± 42.62 pg/30 s, 487.62 ± 49.21 pg/30 s) were significantly higher than those of the chronic periodontitis group (268.81 ± 28.64 pg/30 s, 423.65 ± 35.24 pg/30 s), the gingivitis group (146.35 ± 17.46 pg/30 s, 310.24 ± 48.20 pg/30 s) and the periodontally healthy group (92.46 ± 22.04 pg/30 s, 148.41 ± 24.64 pg/30 s). Similarly, the total levels of IL‐1β and TNF‐α in the generalized aggressive periodontitis group (110.23 ± 9.20 pg/30 s, 1284.46 ± 86.32 pg/30 s) were significantly higher than those in the chronic periodontitis group (423.65 ± 35.24 pg/30 s, 82.64 ± 9.12 pg/30 s), the gingivitis group (52.10 ± 7.15 pg/30 s, 824.24 ± 44.68 pg/30 s) and the periodontally healthy group (36.44 ± 8.86 pg/30 s, 628.26 ± 34.61 pg/30 s). Conclusion: CCL28, IL‐8, IL‐1β and TNF‐α may play key roles in the host response to inflammation in periodontal diseases. As the severity of periodontal diseases increases, destruction of periodontal tissues also increases. Inflammation is one among many factors that trigger periodontal tissue destruction. Identification of the mediators that influence the development and progression of inflammation in periodontal diseases may be very important in understanding the prognoses of periodontal diseases.