Immune response characterization of primary gingival fibroblasts from Grade C periodontitis patients.

BACKGROUND: Grade C, Stage 3-4 Periodontitis (Perio4C) is a rapidly destructive disease caused by an unequilibrated immune response that starts after the primary contact of the periodontopathogens with the gingival tissue. However, it is still unclear how this imbalanced response initiates and what is the role of the connective tissue cells in the progression of this disease. Thus, this study aims to assess the local immune response of Perio4C patients through the exposure of primary gingival fibroblast cells (GFs) with Aggregatibacter actinomycetemcomitans protein extract (AaPE) and the quantification of the inflammatory cytokines interleukin (IL)-4, IL-17, tumor necrosis factor (TNF)-α, IL-1ß, interferon (IFN)-γ, and IL-10 super-family members (IL-10, IL-19, and IL-24) secreted by them. METHODS: Gingival biopsies from nine periodontally health (PH) and eight Perio4C patients were harvested, and the primary culture of GFs was obtained. The cells were exposed to AaPE (5 and 20 µg/ml) and 12-myristate 13-phorbol acetate and ionomycin - calcium salt (PMA). The supernatant was collected after 1.5 and 3 h, and a cytokine panel was evaluated. RESULTS: Clustering analysis indicated dissimilar and stimuli-dependent cytokine production between Perio4C and PH subjects. Perio4C GFs presented lower production of IL-4, TNF-α, IFN-γ, IL-17, IL-10, IL-24, and IL-19, while IL-1ß levels were similar to the PH group, leading to a disruption in the pro-/anti-inflammatory cytokine ratio (p < 0.05). IL-1ß and IL-10 super-family were the most discriminative representants for PH and Perio4C, respectively. CONCLUSION: GFs from individuals with Perio4C tended to hypo-respond to stimulation with AaPE, producing lower concentrations of some pro- and anti-inflammatory molecules, trending to develop a pro-inflammatory extracellular environment.

Effects of the Pulsed Electromagnetic Fields on Experimental Periodontitis and Estrogen Deficiency.

Periodontitis is an inflammatory disease resulting from a complex polymicrobial infection that causes tissue destruction in susceptible individuals. Osteoporosis has been associated with greater clinical attachment loss in patients with periodontitis. Experimental studies have shown positive results in the treatment of osteoporosis through pulsed electromagnetic field (PEMF) stimulation. The aim of this study was to evaluate the effects of PEMF in the presence of estrogen deficiency associated with periodontitis, verifying its role in bone metabolism and in the inflammatory response. Sixty rats were divided into four groups: Sham surgery + ligature-induced periodontitis (P); Sham surgery + ligature-induced periodontitis + PEMF therapy (P + PEMF); Ovariectomy surgery + ligature-induced periodontitis (P + OVX); Ovariectomy surgery + ligature-induced periodontitis + PEMF therapy (P + OVX + PEMF). The area of bone loss in the furcation region (BL), connective tissue attachment loss (CTAL) and alveolar bone loss (ABL), BV/TV and BMD were evaluated. In addition to immunohistochemical labelling of RANKL, OPG, and TRAP and the inflammatory response of interleukin (IL)-1b, IL-6, TNF-α, IL-10, and vascular endothelial growth factor. P + OVX showed significant BL in relation to P + PEMF and the greatest CTAL and ABL. P + OVX and P + OVX + PEMF showed a significant reduction in BV/TV (%). P and P + PEMF showed a significantly lesser amount of Tb.Sp (mm) while P + OVX and P + OVX + PEMF showed a lesser of Tb.N. P + PEMF had the greatest BMD. P + OVX presented higher RANKL and lower OPG immunolabeling than other groups. P + PEMF and P + OVX + PEMF showed a reduction on all biomarkers evaluated. The application of PEMF seems to attenuate the effects of bone loss in the presence of periodontitis and ovariectomy. © 2022 Bioelectromagnetics Society.

IL10 promoter rs6667202 polymorphism is functional in health but not in grade c periodontitis patients: A pilot study.

BACKGROUND AND OBJECTIVE: Previous studies have demonstrated an association between the IL10 promoter rs6667202 (C > A) single-nucleotide polymorphism (SNP) and grade C, stage 3 or 4 periodontitis (Perio4C) in the Brazilian population, where the altered A allele was detected more frequently in these patients. However, no functional analysis of this variation has yet been performed. Thus, the objective of this preliminary study was to evaluate the functionality of rs6667202 in gingival fibroblasts (GFs) of individuals with Perio4C and with periodontal health (PH) stimulated with Aggregatibacter actinomycetencomitans protein extract (AaPE). METHODS: Patients with PH and Perio4C were segregated according to their genotype (AA, AC, or CC), and a biopsy was performed to establish the culture of the GFs. After GFs exposure to AaPE at 5 µg/ml for 1.5 h, RNA was extracted to analyze IL10 expression by qPCR. Aliquots of the cell's supernatant were subjected to immunoenzymatic analysis (MAGpix) to detect interleukin-10 (IL-10). RESULTS: In PH, the genotypes AA and AC are related to less expression of IL10 (p = 0.027 and p < 0.0001) and less production of IL-10 (p = 0.002 and p = 0.001), when compared to CC. In Perio4C, there was no statistical difference between the genotypes (p > 0.05), although a lower IL-10 expression and release compared with PH CC was seen (p = 0.033 and p < 0.001). CONCLUSION: The rs6667202 SNP is functional in PH, as it decreases the expression and production of IL-10. In Perio4C, other factors may be masking its action by altering the IL-10's response.

Connective tissue graft associated or not with low laser therapy to treat gingival recession: randomized clinical trial.

BACKGROUND: To evaluate the treatment of gingival recession with a connective tissue graft (CTG) alone or in combination with low-level laser therapy (CTG + L). METHODS: Forty patients presenting 40 Miller Class I and II gingival recessions were included. The defects were randomly assigned to receive either CTG (n = 20) or CTG + L (n = 20). A diode laser (660 nm) was applied to the test sites immediately after surgery and every other day for 7 days (eight applications). RESULTS: The mean percentage of root coverage was 91.9% for the test group and 89.48% for the control group after 6 months (p > 0.05). The test group presented more complete root coverage (n = 13, 65%) than the control group (n = 7, 35%) (p = 0.04). Dentine sensitivity decreased significantly after 6 months in both groups (p < 0.001). The two groups showed improvement in aesthetics at the end of treatment. CONCLUSIONS: Low-level laser therapy may increase the percentage of complete root coverage when associated with CTG.