Lorenzo Bevilacqua

Caries and Innate Immunity: DEFB1 Gene Polymorphisms and Caries Susceptibility in Genetic Isolates from North-Eastern Italy

Background: The DEFB1 gene, encoding for the constitutively expressed human β-defensin 1 (hBD1) antimicrobial peptide is a potential candidate when studying genetic susceptibility to caries. DEFB1 genetic variations have been reported as contributing to hBD1 production impairment, leading to a greater susceptibility to be infected by oral pathogens, also leading to periodontitis. Methods: We analysed 5 DEFB1 polymorphisms, namely 3 functional single-nucleotide polymorphisms (SNPs) at the 5′-untranslated region (UTR), -52G>A (rs1799946), -44C>G (rs1800972), and -20G>A (rs11362), 2 SNPs at the 3′-UTR, c*5G>A (rs1047031) and c*87A>G (rs1800971) SNP located in potential miRNA binding sites, looking for possible correlations with the risk to develop caries in 654 adult subjects from isolated populations of north-eastern Italy. Dental caries prevalence was evaluated with the DMFT (decayed, missing, filled teeth) index, calculated after an accurate oral examination. DEFB1 SNP genotyping was performed with an Illumina 370k high-density SNP array. Results: Two DEFB1 SNPs were significantly associated with the DMFT index: the strongest association emerged from rs11362 SNP (p = 0.008). In particular G/G homozygous individuals showed a higher DMFT index compared to both G/A heterozygous and A/A homozygous individuals; rs1799946 SNP was also significantly associated with DMFT (p = 0.030), and individuals homozygous for the T allele had a higher DMFT value compared to heterozygous C/T and homozygous C/C individuals. Conclusions: Our study replicated, on a larger number of individuals, previous findings showing the association between two 5′-UTR SNPs in the DEFB1 gene and DMFT, suggesting that these polymorphisms could be considered as potential markers for assessing the risk to develop caries.

Biofilms Developed on Dental Implant Titanium Surfaces with Different Roughness: Comparison Between In Vitro and In Vivo Studies

Microbial biofilms developed on dental implants play a major role in perimplantitis’ pathogenesis. Many studies have indicated that surface roughness is the main feature favoring biofilm development in vitro, but its actual influence in vivo has still to be confirmed. In this study, the amount of biofilm formed on differently treated titanium surfaces, showing distinct roughness, has been examined both in vivo and in vitro by Confocal Laser Scanning Microscopy. In vitro studies availed of biofilm developed by Pseudomonas aeruginosa or by salivary bacteria from volunteer donors. In vivo biofilm production was obtained by exposing titanium discs to the oral cavity of healthy volunteers. In vitro experiments showed that P. aeruginosa and, to a lesser extent, salivary bacteria produce more biomass and develop thicker biofilms on laser-treated and sandblasted titanium surfaces with respect to machined ones. In vivo experiments confirmed that bacterial colonization starts on sites of surface unevenness, but failed to disclose biomass differences among biofilms formed on surfaces with different roughness. Our study revealed that biofilm developed in vitro is more easily influenced by surface features than biofilm formed by complex communities in the mouth, where the cooperation of a variety of bacterial species and the presence of a wide range of nutrients and conditions allow bacteria to optimize substrate colonization. Therefore, quantitative differences observed in vitro among surfaces with different characteristics may not be predictive of different colonization rates in vivo.

Scoring systems for Oral Lichen Planus used by differently experienced raters

Background Scoring systems have been widely used to evaluate the severity and activity of oral lichen planus (OLP). The aim of the present study was to compare two existing (one modified) scoring systems in the evaluation of OLP severity and correlation with pain. Three differently experienced raters were involved. Material and Methods Consecutive patients with OLP were assessed for pain using the Visual Analogue Scale and examined at 10 intraoral sites before starting (T0) and three weeks after (T1) steroid therapy (Clobetasol). Three differently experienced raters evaluated photographs using two scoring systems designated White-Erosive-Atrophic (WEA) modified from an older WEA system (WEA-MOD) and Reticular-erythematous-Ulcerative (REU) systems. WEA-MOD Kendall’s W and interclass correlation coefficient were calculated and correlation between REU/WEA-MOD and pain was calculated using Spearman coefficient. Results Most patients showed lesions on buccal mucosa (85-93,5%) and maxillary/mandibular gingivae (31,8-31,2%), predominantly reticular. At T0, Kendall-W coefficients of 0.89 and 0.74 were obtained for the REU and WEA respectively. At T1, Kendall-W coefficients of 0.83 and 0.58 were obtained for the REU and WEA respectively. Interclass correlation coefficient ranged from 0.87 to 0.90 for REU and from 0.58 to 0.87 for WEA. REU and WEA scores significantly decreased after therapy (p<0.000) as well as VAS (p<0.05). REU score showed correlation with VAS. Conclusions All the raters achieved comparable measures using REU whereas WEA and WEA-MOD seem less reproducible. REU seems to correlate to disease activity and pain. Key words:Oral lichen planus, scoring system, VAS, REU, WEA, rater.

Volumetric Analysis of Gingival Crevicular Fluid and Peri-Implant Sulcus Fluid in Healthy and Diseased Sites: A Cross-Sectional Split-Mouth Pilot Study.

Background: Researchers have recently drawn attention to the analysis of gingival crevicular fluid (GCF) and peri-implant sulcus fluid (PISF) for the implementation of the diagnosis of periodontal and peri-implant disease. Nevertheless, the measurements of volume and biomarkers concentration can be critically biased when data collected from studies with parallel group design are compared, given the technical difficulties, methodological variables, as well as the variability of crevicular fluid characteristics among different individuals. Objective: The aim of the present study was to assess the GCF and PISF volumes in healthy and diseased sites belonging to the same patient. Method: Ten patients presenting a periodontally healthy tooth, a tooth with periodontitis, an implant with healthy peri-implant tissues and an implant with peri-implantitis were enrolled. Samples of GCF and PISF were collected from each site of interest and their volume measured with a Periotron 8000 device. Non-parametric statistical analysis was performed to test the significance of the differences in GCF and PISF volumes between i) sites of teeth and dental implants with the same condition of health or disease and ii) healthy and diseased sites of both teeth and dental implants subgroups. The correlation between probing pocket depth (PPD) and fluid production was also tested (p<0.05). Results: Healthy periodontal and peri-implant tissues produced comparable amounts of fluid that was significantly lower than in diseased sites (p<0.05). In the presence of diagnosed disease, the volumes of GCF and PISF were similar, too. The correlation between PPD and fluid production was significant only in healthy sites (PPD/GCF, ρ=0.890, p<0.001; PPD/PISF, ρ=0.810; p<0.005). Conclusion: The periodontal and peri-implant tissues behaved similarly in terms of fluid production in condition of both health and active disease.