Normal wound healing occurs in four overlapping stages - hemostasis, inflammation, proliferation, and remodeling. In the oral cavity, these processes occur in an infectious environment. Among the pathogens found in the oral community, Aggregatibacter actinomycetemcomitans (Aa) constitutes a well-recognized periodontal pathogen responsible for expressing several virulence factors, which activate a host response. Aim This study investigated whether Aa’s presence can interfere with oral keratinocyte tissue healing in an in vitro wound healing model. Methods Two groups were defined: Group KO (n=5) and Group KO+Aa (n=5). The Aa (JP2 strains) were cultivated in anaerobiosis, and the total protein extract was obtained. The keratinocytes were cultivated with the medium of standard culture until their confluence. After confluence, plates were allocated to each group. With the pipette’s tip, a “scratch” was made in the middle of each well of the plate, and the cells were cultured at 37°C in a humidified atmosphere with 5% CO2. The cells received the stimulus according to groups, and, at times 0, 5, 10, 24, and 48 hours, the wound areas were visualized and standardly recorded using an inverted microscope. Results When analyzing the timeframe, differences in wound measurements indicate a faster closure in the control group compared to the KO+Aa group, although not statistically significant. However, upon examining the wound closure measures, it was observed that the Aa protein extract significantly reduced wound closure at 10 and 48 hours (p<0.05), negatively impacting the keratinocyte’s behavior. Conclusion In summary, it was demonstrated that the pathogen Aa can interfere with the re-epithelization in vitro.

Abstract This study aimed to assess the influence of smoking on the subgingival metatranscriptomic profile of young patients affected by stage III/IV and generalized periodontal disease. Methodology In total, six young patients, both smokers and non-smokers (n=3/group), who were affected by periodontitis were chosen. The STROBE (Strengthening the Reporting of Observational Studies in Epidemiology) guidelines for case-control reporting were followed. Periodontal clinical measurements and subgingival biofilm samples were collected. RNA was extracted from the biofilm and sequenced via Illumina HiSeq. Differential expression analysis used Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment, and differentially expressed genes were identified using the Sleuth package in R, with a statistical cutoff of ≤0.05. Results This study found 3351 KEGGs in the subgingival biofilm of both groups. Smoking habits altered the functional behavior of subgingival biofilm, resulting in 304 differentially expressed KEGGs between groups. Moreover, seven pathways were modulated: glycan degradation, galactose metabolism, glycosaminoglycan degradation, oxidative phosphorylation, peptidoglycan biosynthesis, butanoate metabolism, and glycosphingolipid biosynthesis. Smoking also altered antibiotic resistance gene levels in subgingival biofilm by significantly overexpressing genes related to beta-lactamase, permeability, antibiotic efflux pumps, and antibiotic-resistant synthetases. Conclusion Due to the limitations of a small sample size, our data suggest that smoking may influence the functional behavior of subgingival biofilm, modifying pathways that negatively impact the behavior of subgingival biofilm, which may lead to a more virulent community.

Abstract Periodontitis Stage III-IV, Grade C (PerioC) is a severe form of Periodontitis. The individual genetic background has been shown to be an important etiopathogenic factor for the development of this disease in young, systemically healthy, and non-smokers patients. Recently, after exome sequencing of families with a history of the disease, PerioC was associated with three single nucleotide variations (SNVs) – rs142548867 (EEFSEC), rs574301770 (ZNF136), and rs72821893 (KRT25) – which were classified as deleterious or possibly harmful by prediction algorithms. Objective Seeking to validate these findings in a cohort evaluation, this study aims to characterize the allele and genotypic frequency of the SNVs rs142548867, rs574301770, and rs72821893 in the Brazilian population with PerioC and who were periodontally healthy (PH). Methodology Thus, epithelial oral cells from 200 PerioC and 196 PH patients were harvested at three distinct centers at the Brazilian Southern region, their DNA were extracted, and the SNVs rs142548867, rs574301770, rs72821893 were genotyped using 5′-nuclease allelic discrimination assay. Differences in allele and genotype frequencies were analyzed using Fisher’s Exact Test. Only the SNV rs142548867 (C > T) was associated with PerioC. Results The CT genotype was detected more frequently in patients with PerioC when compared with PH subjects (6% and 0.5% respectively), being significantly associated with PerioC (odds ratio 11.76, p=0.02). Conclusion rs142548867 represents a potential risk for the occurrence of this disease in the Brazilian population.