Gene expression of inflammatory immune factors and clinical parameters in diabetes and nondiabetes patients with periodontal disease

Objective: This study evaluated clinical, glucose, and immunological parameters in patients with type 2 diabetes mellitus (DM) compared to those without systemic alterations (NDM), both with generalized chronic periodontitis. Methodology: Twenty-one patients were selected with indications of tooth loss. Surgeries were performed using the Widman flap modified to obtain a gingival collar at 1 mm from the gingival margin. Before the surgical procedure, the following clinical parameters were evaluated: pocket probing depth (PPD), gingival recession (GR), clinical attachment level (CAL), and bleeding on probing (BOP). Fasting glucose levels (FGL) and glycosylated hemoglobin Hba1c (HbA1c) were also assessed. During the surgery, gingival tissue samples were collected and frozen for later laboratorial analysis. The samples were processed to obtain mRNA, cDNA and determine the gene expression of the immune parameters IL-1-β, IL-6, TNF-α, IL-10 and NF-kB by real-time polymerase chain reaction (RT-PCR). Data were analyzed statistically considering p<0.05. Results: The clinical and glucose parameters BOP, FGL, and HbA1c were statistically higher in the DM group. RNAm levels of IL-1β, TNF-α, and NF-kB were higher in the DM group (p<0.05). Conclusion: The presence of diabetes and hyperglycemic status increase the levels of pro-inflammatory immune factors and severity of the periodontal disease.

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Copper decreases gene expression of TNF-α, IL-10, and of matrix metalloproteinases MMP-2 and MMP-9 in isolated perfused rat livers

Biologia ◽

10.2478/s11756-007-0061-0 ◽

2007 ◽

Vol 62 (3) ◽

Cited By ~ 2

Author(s):

Albena Alexandrova ◽

Elena Bandžuchová ◽

Anton Kebis ◽

Marián Kukan ◽

Daniel Kuba

Keyword(s):

Oxidative Stress ◽

Gene Expression ◽

Matrix Metalloproteinase ◽

Oxidative Dna Damage ◽

Interleukin 10 ◽

Matrix Metalloproteinase 2 ◽

Necrosis Factor Alpha ◽

Tissue Samples ◽

Tnf Α ◽

Rat Livers

AbstractCopper is known to induce oxidative stress in a number of models. It was shown that many pathophysiological events were associated with oxidative stress. Further, oxidative stress can increase gene expression of cytokines and of metalloproteinases. We previously found that copper toxic effects in isolated perfused rat livers were associated with significant oxidative stress (as assessed by lipid peroxidation, protein oxidation and oxidative DNA damage, particularly at concentration of 0.03 mM of Cu2+ in the perfusate). Here we investigated gene expression of tumor necrosis factor-alpha (TNF-α), interleukin-10 (IL-10); matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in frozen liver tissue samples by the real-time PCR assay. Compared to controls, copper at concentration of 0.01 mM did not affect gene expression of TNF-α, IL-10, MMP-2 and MMP-9, whereas copper at concentration of 0.03 mM significantly decreased gene expression of all the four TNF-α, IL-10, MMP-2 and MMP-9 by 69%, 81%, 43%, and 62%, respectively. These results suggest that copper-induced oxidative stress in the isolated rat liver can lead to the suppression of gene expression. Because TNF-α and metalloproteinases are involved also in liver regeneration, the suppression of these genes by copper may be one of the mechanisms by which acute intoxication of animals and humans with copper may impair regenerative capability of the liver.

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Effect of Non-Surgical Periodontal Therapy on Superoxide Dismutase Levels in Gingival Tissues of Chronic Periodontitis Patients: A Clinical and Spectophotometric Analysis

Disease Markers ◽

10.1155/2013/269027 ◽

2013 ◽

Vol 34 (5) ◽

pp. 305-311 ◽

Cited By ~ 4

Author(s):

Laxmi Sukhtankar ◽

Anita Kulloli ◽

Rahul Kathariya ◽

Sharad Shetty

Keyword(s):

Superoxide Dismutase ◽

Chronic Periodontitis ◽

Inflammatory Cells ◽

Periodontal Therapy ◽

Periodontal Pocket ◽

Gingival Tissue ◽

Control Group ◽

Clinical Parameters ◽

Sod Activity ◽

Tissue Samples

BACKGROUND: Superoxide dismutase (SOD), an antioxidant acting against superoxide (oxygen radical, O2.-), it is released in inflammatory pathways and causes connective tissue breakdown. Increased SOD activity in inflamed gingiva may indicate increased O2.-radical generation by neutrophils and other inflammatory cells at the diseased site. The aim of the study was to evaluate the effects of non-surgical periodontal therapy (NSPT) on SOD levels in gingival tissues of chronic periodontitis patients.METHODS: Forty subjects: 20 periodontally healthy (Control) and 20 chronic periodontitis (Test); age range 24–55 years were recruited. Gingival tissue samples were collected by excising the inner lining of the periodontal pocket at baseline (prior to non-surgical periodontal therapy) and 2 months post therapy. In controls, tissue samples were obtained immediately after tooth extraction scheduled for orthodontic reasons. Clinical parameters included probing depth, clinical attachment level, gingival index, bleeding index, plaque index. SOD activities were assessed spectrophotometrically at baseline and 2 months post NSPT, results were analysed statistically.RESULTS: At baseline, patients with chronic periodontitis had higher mean SOD activity (2.73 ± 1.36) than the control subjects (1.12 ± 1.13) withp= 0.00003 (p< 0.05). At 2 months post NSPT median SOD level (1.00) had come close to median SOD value of control group (0.85);p= 0.99 (p> 0.05). The resolution of inflammation with successful NSPT resulted in decreased SOD levels as in control group. Clinical parameters in patients with chronic periodontitis showed a significant improvement 2 months post NSPT (p< 0.05).CONCLUSION: Non-surgical periodontal therapy significantly improves the clinical parameters and restores previously increased SOD levels to normal in chronic periodontitis patients.

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Expression of deubiquitinases in human gingiva and cultured human gingival fibroblasts

BMC Oral Health ◽

10.1186/s12903-021-01655-4 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Yong-Wei Fu ◽

Hong-Zhi Xu

Keyword(s):

Protein Expression ◽

Human Gingival Fibroblasts ◽

Gingival Tissue ◽

Gingival Fibroblasts ◽

Western Blots ◽

Tissue Samples ◽

Periodontal Tissues ◽

Tnf Α ◽

Gene And Protein Expression ◽

Human Gingiva

Abstract Background Although deubiquitinating enzymes (DUBs) such as CYLD, A20 and OTULIN are expressed in multiple tissues and thought to be linked with inflammatory diseases, their expression in periodontal tissues remains to be determined. This research was designed to assess the expression of CYLD, A20 and OTULIN in human gingiva, and to evaluate the regulation of these DUBs in human gingival fibroblasts (HGFs) upon different stimuli. Methods Immunohistochemistry assay was conducted to determine the expression of CYLD, A20 and OTULIN in human gingiva. Immunofluorescence assay was employed to observe the protein expression of CYLD, A20 and OTULIN in HGFs. RT-PCR and western blots were carried out to assess gene and protein expression changes of these DUBs in HGFs upon LPS or TNF-α. Results CYLD, A20 and OTULIN were found to be expressed in human gingiva and HGFs. The expression of CYLD, A20 and OTULIN was lower in the inflamed gingival tissue samples compared with the healthy gingival tissue samples. Further, the expression of CYLD, A20 and OTULIN in HGFs exhibited distinct regulation by different stimuli. TNF-α treatment markedly increased NF-κB activation in HGFs Conclusions Our findings suggest that CYLD, A20 and OTULIN might play a role in the progression of periodontitis.

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The Molecular Analysis of rs11614913 Polymorphism from miRNA196a Gene and Its Relationship with TNF-α Gene Expression in Cervical Cancer

Jentashapir Journal of Cellular and Molecular Biology ◽

10.5812/jjcmb.101796 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Ahmad Hamta ◽

Fatemeh Hajihassani

Keyword(s):

Gene Expression ◽

Cervical Cancer ◽

Malignant Tumors ◽

Control Group ◽

Tissue Samples ◽

Altered Expression ◽

Tnf Α ◽

Cancer Sample ◽

Rs11614913 Polymorphism ◽

Factor Α

Background: Cervical cancer (CC) is one of the most common malignant tumors in women, which has been diagnosed as fourth cancer in females worldwide. In addition to human papillomavirus (HPV), genetic factors, including altered expression of some microRNAs and mutations in tumor necrosis factor α (TNF-α) gene, are involved in this cancer. Objectives: This study aimed to investigate the rs11614913 polymorphism from the miRNA196a gene and its association with the expression of the TNF-α gene in cervical cancer for early diagnosis and treatment. Methods: In this study, 52 samples of pre-cancerous and cancerous lesions, and 50 tissue samples were collected from healthy subjects in an Iranian population. DNA was extracted from the samples, and rs11614913 polymorphism of the miRNA196a gene was investigated by PCR. RNA was extracted from the samples, and the expression of the miRNA196a and TNF-α genes were evaluated. Finally, for data analysis, Epi Info software version 7.1.3.10 and MedCalc Version 19.2.0 were used. Results: The frequency of CC, TC, and TT genotypes from rs11614913 polymorphism of miRNA196a gene was 0.58, 0.34, and 0.08, respectively, but in the healthy group it was 0.36, 0.46, and 0.18, respectively. The results also showed that the expression of miRNA196a and TNF-α genes in the patient group was higher than the control group. Conclusions: Based on the results of this study, a significant correlation was found between CC genotype and rs11614913 polymorphism of miRNA196a gene and TNF-α gene expression in the cervical cancer sample. Therefore, investigating these factors in patients with cervical cancer may be helpful.

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Levels of Gene Expression of Immunological Biomarkers in Peri-Implant and Periodontal Tissues

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph17239100 ◽

2020 ◽

Vol 17 (23) ◽

pp. 9100

Author(s):

Luciene Cristina Figueiredo ◽

Bruno Bueno-Silva ◽

Cristiana Fernandes Plutarco Nogueira ◽

Leonardo Carneiro Valadares ◽

Katia Marina Morilla Garcia ◽

...

Keyword(s):

Gene Expression ◽

Matrix Metalloproteases ◽

Gingival Tissue ◽

Mrna Levels ◽

Necrosis Factor Alpha ◽

Periodontal Tissues ◽

Tnf Α ◽

The Matrix ◽

Study Population ◽

Factor Alpha

This study compared the gene expression of the immunoinflammatory markers interleukin (IL)-6, IL-1ß, and tumor necrosis factor alpha (TNF-α), the matrix metalloproteinases (MMP)-1, -2, -8, and -9, and the tissue inhibitors of matrix metalloproteases (TIMP)-1 and -2 in the gingival tissue of individuals with periodontal and peri-implant disease. The study population included individuals with four periodontal statuses: periodontal health (PH group, n = 20); periodontitis (P group, n = 20); peri-implant health (PIH group, n = 20), and peri-implantitis (PI group, n = 20). Gingival biopsies were collected from one tooth per patient according to the inclusion criteria of each group. The mRNA levels of IL-6, IL-1ß, TNF-α, MMP-1, MMP-2, MMP-8, MMP-9, TIMP-1, and TIMP-2 were evaluated by qPCR. The levels of IL-1ß were significantly higher in the PI group when compared to the other groups (p < 0.05), while the levels of IL-6 were significantly higher in the groups with periodontal and peri-implant disease when compared with the healthy groups (p < 0.05); however, the levels of IL-6 did not differ between the PI and P groups (p > 0.05). For all other studied biomarkers, no significant differences were observed between groups (p > 0.05). IL-6 and IL-1ß presented higher levels of mRNA in diseased periodontal and peri-implant tissues. However, the expression of metalloproteinases and their inhibitors did not differ between the different periodontal statuses.

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Immunoexpression of HIF-1α and VEGF in Periodontal Disease and Healthy Gingival Tissues

Brazilian Dental Journal ◽

10.1590/0103-6440201600533 ◽

2016 ◽

Vol 27 (2) ◽

pp. 117-122 ◽

Cited By ~ 21

Author(s):

Roseane Carvalho Vasconcelos ◽

Antônio de Lisboa Lopes Costa ◽

Roseana de Almeida Freitas ◽

Bruna Aguiar do Amaral Bezerra ◽

Bruna Rafaela Martins dos Santos ◽

...

Keyword(s):

Endothelial Cells ◽

Periodontal Disease ◽

Hypoxia Inducible Factor ◽

Gingival Tissue ◽

Vegf Expression ◽

Median Percentage ◽

Tissue Samples ◽

Inflammatory Conditions ◽

Bacterial Endotoxins ◽

And Migration

Abstract Hypoxia-inducible factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) are proteins that stimulate the proliferation and migration of endothelial cells. These proteins have been described in many pathologic and inflammatory conditions, but their involvement in the development of periodontitis has not been thoroughly investigated. This study compared the immunohistochemical expression of these proteins, involved in angiogenesis and hypoxia, by imunnostained inflammatory and endothelial cells in periodontal disease and healthy gingival tissues. Gingival tissue samples were divided as follows: 30 samples with chronic periodontitis, 30 with chronic gingivitis, and 30 of healthy gingiva. Results were analyzed statistically by the Kruskal-Wallis, Mann-Whitney and Spearman correlation tests (p=0.01). Inflammatory and endothelial cells were found to express these proteins. Periodontitis showed median percentage of HIF-1α-positive cells of 39.6%, 22.0% in cases of gingivitis and 0.9% in the healthy gingiva group (p=0.001). For VEGF, median percentage of immunopositive cells was 68.7% for periodontitis, 66.1% in cases for gingivitis, and 19.2% for healthy gingival specimens (p<0.001). Significant correlation between VEGF and HIF-1α was also observed in healthy gingiva (p<0.001).The increased expression of HIF-1αα and VEGF in periodontitis, compared to gingivitis and healthy gingiva, suggests possible activation of the HIF-1α pathway in advanced periodontal disease. The correlation between HIF-1α and VEGF expression in healthy gingiva suggests a physiological function for these proteins in conditions of homeostasis. In periodontal disease, HIF-1 and VEGF expression may be regulated by other factors, in addition to hypoxia, such as bacterial endotoxins and inflammatory cytokines.

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Effect of Periodontal Surgery on Tissue Plasminogen Activator and Plasminogen Activator Inhibitor Type-1 Gene Expression in Gingival Tissues of Periodontitis Patients: A Controlled Before-And-After Study

10.26810/perioj.2019.a2 ◽

2019 ◽

Vol 3 (1) ◽

pp. 7-17

Author(s):

Sandy Hassan ◽

Iman Aboushady ◽

Gihane Madkour

Keyword(s):

Gene Expression ◽

Plasminogen Activator ◽

Plasminogen Activator Inhibitor ◽

Gingival Tissue ◽

Control Group ◽

Tissue Samples ◽

Expression Levels ◽

Before And After ◽

Activator Inhibitor Type ◽

Pai 1

Background: Regulation of the plasminogen activation system (PAS) is a vital component in governing proteolytic events within the extracellular matrix (ECM). PAS is believed to play a substantial role in the destruction and healing of periodontal tissues. Thus, the current work aimed to study the histopathological effect of open flap debridement (OFD) on periodontitis, as well as its effect on tissue plasminogen activator (t-PA) and plasminogen activator inhibitor type-1 (PAI-1) gene levels in gingival tissues. Methods: A total of 30 subjects were enrolled in the present study. They were divided into two groups: Group I (control group) included 10 periodontally healthy volunteers and group II (periodontitis group) comprised 20 patients suffering from stage III grade B periodontitis. Gingival tissue samples were collected from all periodontitis patients, before and after OFD, and from healthy controls. Hematoxylin and eosin (H&E) stained slides were subsequently examined and gene expression levels of t-PA and PAI-1 were assessed in the gingiva through quantitative reverse transcription polymerase chain reaction (RT-PCR). Results: Gingival tissue samples from periodontitis patients showed widely dilated blood vessels, diffuse hemorrhage, areas of edema, and disorganized collagen fibers together with large amounts of inflammatory cells in between. Following OFD, smaller sized blood vessels, a restored collagen fiber distribution, and an obvious decrease in the inflammatory infiltrate were noted. Gene expression levels of t-PA and PAI-1 were significantly higher in the periodontitis patients compared to the healthy controls. Although their levels showed a significant decrease following OFD in the periodontitis group, they were still significantly higher than the control group. Conclusion: OFD procedures resulted in down regulation of t-PA and PAI-1 expression levels in the gingiva of periodontitis patients, which could signify an important role of these proteins on periodontal disease progression.

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Dynamics of Acute Local Inflammatory Response after Autologous Transplantation of Muscle-Derived Cells into the Skeletal Muscle

Mediators of Inflammation ◽

10.1155/2014/482352 ◽

2014 ◽

Vol 2014 ◽

pp. 1-12 ◽

Cited By ~ 7

Author(s):

Anna Burdzinska ◽

Kamila Gala ◽

Cezary Kowalewski ◽

Radosław Zagozdzon ◽

Zdzisław Gajewski ◽

...

Keyword(s):

Gene Expression ◽

Skeletal Muscle ◽

Inflammatory Response ◽

Immune Reaction ◽

Mrna Level ◽

Autologous Transplantation ◽

Inflammatory Infiltration ◽

Tissue Samples ◽

Local Inflammatory Response ◽

Tnf Α

The vast majority of myoblasts transplanted into the skeletal muscle die within the first week after injection. Inflammatory response to the intramuscular cell transfer was studied in allogeneic but not in autologous model. The aim of this study was to evaluate immune reaction to autotransplantation of myogenic cells and to assess its dynamics within the first week after injection. Muscle-derived cells or medium alone was injected into the intact skeletal muscles in autologous model. Tissue samples were collected 1, 3, and 7 days after the procedure. Our analysis revealed the peak increase of the gene expression of all evaluated cytokines (Il-1α, Il-1β, Il-6, Tgf-β,andTnf-α) at day 1. The mRNA level of analyzed cytokines normalized in subsequent time points. The increase ofIl-βgene expression was further confirmed at the protein level. Analysis of the tissue sections revealed rapid infiltration of injected cell clusters with neutrophils and macrophages. The inflammatory infiltration was almost completely resolved at day 7. The survived cells were able to participate in the muscle regeneration process. Presented results demonstrate that autotransplanted muscle-derived cells induce classical early immune reaction in the site of injection which may contribute to cellular graft elimination.

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Decreased TNF-α gene expression in periodontal ligature in MSG-obese rats: A possible protective effect of hypothalamic obesity against periodontal disease?

Archives of Oral Biology ◽

10.1016/j.archoralbio.2011.08.024 ◽

2012 ◽

Vol 57 (3) ◽

pp. 300-306 ◽

Cited By ~ 7

Author(s):

Sávio Brandelero ◽

Maria Lúcia Bonfleur ◽

Rosane Aparecida Ribeiro ◽

Emerielle Cristine Vanzela ◽

Carlos Augusto Nassar ◽

...

Keyword(s):

Gene Expression ◽

Periodontal Disease ◽

Protective Effect ◽

Hypothalamic Obesity ◽

Tnf Α ◽

Obese Rats

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Repetitive Intermittent Hyperglycemia Drives the M1 Polarization and Inflammatory Responses in THP-1 Macrophages Through the Mechanism Involving the TLR4-IRF5 Pathway

Cells ◽

10.3390/cells9081892 ◽

2020 ◽

Vol 9 (8) ◽

pp. 1892 ◽

Cited By ~ 2

Author(s):

Fatema Al-Rashed ◽

Sardar Sindhu ◽

Hossein Arefanian ◽

Ashraf Al Madhoun ◽

Shihab Kochumon ◽

...

Keyword(s):

Gene Expression ◽

Inflammatory Responses ◽

Microvascular Complications ◽

Intensive Therapy ◽

Model Assessment ◽

Tissue Samples ◽

Metabolic Inflammation ◽

M1 Polarization ◽

Tnf Α ◽

Glucose Fluctuations

Repetitive intermittent hyperglycemia (RIH) is an independent risk factor for complications associated with type-2 diabetes (T2D). Glucose fluctuations commonly occur in T2D patients with poor glycemic control or following intensive therapy. Reducing blood glucose as well as glucose fluctuations is critical to the control of T2D and its macro-/microvascular complications. The interferon regulatory factor (IRF)-5 located downstream of the nutrient sensor toll-like receptor (TLR)-4, is emerging as a key metabolic regulator. It remains unclear how glucose fluctuations may alter the IRF5/TLR4 expression and inflammatory responses in monocytes/macrophages. To investigate this, first, we determined IRF5 gene expression by real-time qRT-PCR in the white adipose tissue samples from 39 T2D and 48 nondiabetic individuals. Next, we cultured THP-1 macrophages in hypo- and hyperglycemic conditions and compared, at the protein and transcription levels, the expressions of IRF5, TLR4, and M1/M2 polarization profile and inflammatory markers against control (normoglycemia). Protein expression was assessed using flow cytometry, ELISA, Western blotting, and/or confocal microscopy. IRF5 silencing was achieved by small interfering RNA (siRNA) transfection. The data show that adipose IRF5 gene expression was higher in T2D than nondiabetic counterparts (p = 0.006), which correlated with glycated hemoglobin (HbA1c) (r = 0.47/p < 0.001), homeostatic model assessment of insulin resistance (HOMA-IR) (r = 0.23/p = 0.03), tumor necrosis factor (TNF)-α (r = 0.56/p < 0.0001), interleukin (IL)-1β (r = 0.40/p = 0.0009), and C-C motif chemokine receptor (CCR)-2 (r = 0.49/p < 0.001) expression. IRF5 expression in macrophages was induced/upregulated (p < 0.05) by hypoglycemia (3 mM/L), persistent hyperglycemia (15 mM/L–25 mM/L), and RIH/glucose fluctuations (3–15 mM/L) as compared to normoglycemia (5 mM/L). RIH/glucose fluctuations also induced M1 polarization and an inflammatory profile (CD11c, IL-1β, TNF-α, IL-6, and monocyte chemoattractant protein (MCP)-1) in macrophages. RIH/glucose fluctuations also drove the expression of matrix metalloproteinase (MMP)-9 (p < 0.001), which is a known marker for cardiovascular complication in T2D patients. Notably, all these changes were counteracted by IRF5 silencing in macrophages. In conclusion, RIH/glucose fluctuations promote the M1 polarization and inflammatory responses in macrophages via the mechanism involving TLR4-IRF5 pathway, which may have significance for metabolic inflammation.

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