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.

Open flap debridement compared to repeated applications of photodynamic therapy in the treatment of residual pockets: A randomized clinical trial.

BACKGROUND: The aim of the present study was to compare repeated applications of antimicrobial photodynamic therapy (aPDT) to open flap debridement (OFD) in the treatment of residual periodontal pockets in non-furcation sites. METHODS: Forty-six subjects with a diagnosis of Stage III or IV Grade C periodontitis, that had been previously treated, participated in the study.  Residual pockets were divided between two groups: (1) aPDT group: received ultrasonic periodontal debridement followed by immediate application of aPDT, and repeated on1st, 2nd, 7th, and 14th days; and (2) OFD group: treated by modified papilla preservation technique, where granulation tissue and visible calculus were removed with hand curettes and an ultrasonic device. Clinical, immunological, and microbiological parameters were evaluated before and after treatment. RESULTS: Both treatments were effective reducing clinical parameters of disease. OFD resulted in a greater mean probing pocket depths (PPD) reduction in deep pockets (p = 0.001). However, aPDT resulted in a lower occurrence of gingival recession (GR), dentin hypersensitivity (DH) and analgesic intake. Reduction in Porphyromonas gingivalis was observed in both groups. Only the OFD group had a significant reduction in Aggregatibacter actinomycetemcomitans. aPDT group had greater increase in interleukin 10 (IL-10) levels and a greater reduction of interleukin 1 beta (IL-1ß) at 14 days when compared to the OFD group (p < 0.05). CONCLUSION: OFD was superior in reducing PPD in deep pockets compared to the aPDT. However, OFD resulted in greater GR.  Both treatments lowered P. gingivalis levels but only OFD reduced levels of A. actinomycemtemcomitans.

Childhood Hypophosphatasia Associated with a Novel Biallelic ALPL Variant at the TNSALP Dimer Interface.

The goal of this study was to perform a clinical and molecular investigation in an eight-year-old female child diagnosed with hypophosphatasia (HPP). The proband and her family were evaluated by medical and dental histories, biochemical analyses, radiographic imaging, and genetic analysis of the tissue-nonspecific alkaline phosphatase (ALPL) gene. A bioinformatic analysis was performed to predict the structural and functional impact of the point mutations in the tissue-nonspecific alkaline phosphatase (TNSALP) molecule and to define their potential contribution to the phenotype. We identified a novel combination of heterozygous ALPL missense variants in the proband, p.Ala33Val and p.Asn47His, compatible with an autosomal recessive mode of inheritance and resulting in skeletal and dental phenotypes. Computational modeling showed that the affected Asn47 residue is located in the coil structure close to the N-terminal α-helix, whereas the affected Ala33 residue is localized in the N-terminal α-helix. Both affected residues are located close to the homodimer interface, suggesting they may impair TNSALP dimer formation and stability. Clinical and biochemical follow-up revealed improvements after six years of ERT. Reporting this novel combination of ALPL variants in childhood HPP provides new insights into genotype-phenotype associations for HPP and specific sites within the TNSALP molecule potentially related to a childhood-onset HPP and skeletal and dental manifestations. Beneficial effects of ERT are implicated in skeletal and dental tissues.

Omega-3 PUFA and aspirin as adjuncts to periodontal debridement in patients with periodontitis and type 2 diabetes mellitus: Randomized clinical trial.

BACKGROUND: Supplementation with omega-3 polyunsaturated fatty acids (ω-3 PUFA) and low-dose aspirin (ASA) have been proposed as a host modulation regimen to control chronic inflammatory diseases. The aim of this study was to investigate the clinical and immunological impact of orally administered ω-3 PUFA and ASA as adjuncts to periodontal debridement for the treatment of periodontitis in patients type 2 diabetes. METHODS: Seventy-five patients (n = 25/group) were randomly assigned to receive placebo and periodontal debridement (CG), ω-3 PUFA + ASA (3 g of fish oil/d + 100 mg ASA/d for 2 months) after periodontal debridement (test group [TG]1), or ω-3 PUFA + ASA (3 g of fish oil/d + 100 mg ASA/d for 2 months) before periodontal debridement (TG2). Periodontal parameters and GCF were collected at baseline (t0), 3 months after periodontal debridement and ω-3 PUFA + ASA or placebo for TG1 and CG (t1), after ω-3 PUFA + ASA (before periodontal debridement) for TG2 (t1), and 6 months after periodontal debridement (all groups) (t2). GCF was analyzed for cytokine levels by multiplex ELISA. RESULTS: Ten patients (40%) in TG1 and nine patients (36%) in TG2 achieved the clinical endpoint for treatment (less than or equal to four sites with probing depth ≥ 5 mm), as opposed to four (16%) in CG. There was clinical attachment gain in moderate and deep pockets for TG1. IFN-γ and interleukin (IL)-8 levels decreased over time for both test groups. IL-6 levels were lower for TG1. HbA1c levels reduced for TG1. CONCLUSION: Adjunctive ω-3 and ASA after periodontal debridement provides clinical and immunological benefits to the treatment of periodontitis in patients with type 2 diabetes.

Novel rare frameshift variation in aggressive periodontitis: Exomic and familial-screening analysis.

BACKGROUND: Aggressive periodontitis (AgP), currently periodontitis grade C, presents early onset, rapid progression, and a poorly established genetic association. Thus, this study aimed to identify genetic variants associated with AgP via whole exome sequencing (WES) through a familial screening approach. METHODS: WES was performed in two nuclear families, including a proband and a parent affected by AgP and an unaffected parent and sibling. Common variants among affected individuals, excluding those common to healthy people, from each family, composed the data set associated with AgP. In silico analysis evaluated the impact of each variant on protein structure and protein-protein interactions. Moreover, identified deleterious variants were validated in a populational analysis (n = 96). RESULTS: The missense single nucleotide variations (SNVs) rs142548867 in EEFSEC (c.668C>T), rs574301770 in ZNF136 (c.466C>G), and rs72821893 in KRT25 (c.800G>A) and the frameshift indels rs37146475 in GPRC6A (c.2323-2324insT) and c.1366_1372insGGAGCAG in ELN were identified in AgP and have a predicted functional impact on proteins. In silico analysis indicated that the indel in GPRC6A generates a loss of the C-terminal tail of the Gprca protein. Furthermore, this SNV was significantly associated with AgP in a population-based investigation. CONCLUSION: Novel frameshift variation in GPRC6A (c.2323-2324insT) was identified as a potential genetic alteration associated with AgP occurrence.

Two different antibiotic protocols as adjuncts to one-stage full-mouth ultrasonic debridement to treat generalized aggressive periodontitis: A pilot randomized controlled clinical trial.

BACKGROUND: To assess the clinical and microbiological responses of amoxicillin + metronidazole (AMX + MET) versus clarithromycin (CLM) as adjuncts to one-stage full-mouth ultrasonic debridement (FMUD) in the treatment of generalized aggressive periodontitis (GAgP). METHODS: For this parallel, double-masked, pilot randomized clinical trial, 46 patients with GAgP were selected and randomly assigned into two groups: AMX+MET group (n = 23): FMUD associated with AMX (500 mg three times a day) and MET (400 mg three times a day) for 7 days; and CLM group (n = 23): FMUD associated with CLM (500 mg twice a day) for 7 days. Clinical parameters were evaluated at baseline, 3, and 6 months post-treatment. The levels of Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Fusobacterium nucleatum from subgingival biofilm were determined by quantitative polymerase chain reaction. RESULTS: Both treatments significantly improved all clinical parameters compared with baseline and promoted a significant reduction of A. actinomycetemcomitans and P. gingivalis counts (P > 0.05). CLM succeeded in decreasing T. forsythia at 6 months (P < 0.05), but no antibiotic was able to reduce F. nucleatum. There was no difference between the two protocols regarding the reported adverse effects (P > 0.05). CONCLUSIONS: The results suggest that CLM is not superior than AMX + MET in the treatment of GAgP. However, this antibiotic led to good clinical outcomes and may be a possible alternative to AMX+MET in the treatment of severe periodontitis in young patients. Future studies with larger sample sizes are needed to confirm this statement (NCT02969928).