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Background Statistical methodology is a powerful tool in the health research; however, there is wide accord that statistical methodologies are not usually used properly. In particular when multiple comparisons are needed, it is necessary to check the rate of false positive results and the potential inflation of type I errors. In this case, permutation testing methods are useful to check the simultaneous significance level and identify the most significant factors. Methods In this paper an application of permutation tests, in the medical context of Inflammatory Bowel Diseases, is performed. The main goal is to assess the existence of significant differences between Crohn’s Disease (CD) and Ulcerative Colitis (UC). The Sequentially Rejective Multiple Test (Bonferroni-Holm procedure) is used to find which of the partial tests are effectively significant and solve the problem of the multiplicity control. Results Applying Non-Parametric Combination (NPC) Test for partial and combined tests we conclude that Crohn’s Disease patients and Ulcerative Colitis patients differ between them for most examined variables. UC patients compared with the CD patients, have a higher diagnosis age, not show smoking status, proportion of patients treated with immunosuppressants or with biological drugs is lower than the CD patients, even if the duration of such therapies is longer. CD patients have a higher rate of re-hospitalization. Diabetes is more present in the sub-population of UC patients. Analyzing the Charlson score we can highlight that UC patients have a more severe clinical situation than CD patients. Finally, CD patients are more frequently subject to surgery compared to UC. Appling of the Bonferroni Holm procedure, which provided adjusted p -values, we note that only nine of the examined variables are statistically significant: Smoking habit, Immunosuppressive therapy, Surgery, Biological Drug, Diabetes, Adverse Events, Re-hospitalization, Gender and Duration of Immunosoppressive Therapy. Therefore, we can conclude that these are the specific variables that can discriminate effectively the Crohn’s Disease and Ulcerative Colitis groups. Conclusions We identified significant variables that discriminate the two groups, satisfying the multiplicity problem, in fact we can affirm that Smoking habit, Immunosuppressive therapy, Surgery, Biological Drug, Diabetes, Adverse Events, Hospitalization, Gender and Duration of Immunosoppressive Therapy are the effectively significant variables.

Background Micro-RNAs (miRNAs) have been reported to play an important role during orthodontic tooth movement (OTM) through the regulation of periodontal soft and hard tissue homeostasis and functions. The aim of the present study was to assess the effects of miRNAs on OTM and to evaluate possible predictors that influenced the overall OTM amount at a 3-month follow-up. Methods Through a split-mouth design, 21 healthy patients (mean age 13.2 ± 1.8 years) were enrolled in the present study. Clinical parameters and gingival crevicular fluid (GCF) sampling were performed on both compression and tension sides of a random canine to be distalized (test groups) at baseline and at 1 h, 1 day, 1 month and at 3-month after OTM, while the contralateral canine served as a control group. miRNAs − 7a-3p, -7a-2-3p, -7a-5p, -21-3p, -21-5p, -100-3p, -100-5p, -125b-2-3p, -125b-5p, -200b-3p, and − 200b-5p expression was analyzed using a real-time quantitative polymerase chain reaction (RT-PCR). Data were analyzed to assess miRNAs change following OTM. Spearman test, two-way ANOVA and a multivariate regression model were established to evaluate the correlation among miRNAs and clinical parameters and to explore possible predictors of OTM amount at 3-month follow-up. Results At 3-month follow-up, there was an increase of miRNA-7a-2-3p, -21-5p, -100-5p, a decrease of miRNA-125b-5p, 200b-3p and − 200b-5p in the compression side and an increase of miRNA-7a-3p, 100-5p in the tension side ( p  < 0.05). The two-way ANOVA revealed that OTM determined, on the compression side, a significant upregulation on miRNA-7a-3p ( p  = 0.017), -7a-2-3p ( p  = 0.023), -21-5p ( p  = 0.007), -100-5p ( p  = 0.025) and a significant downregulation of miRNA-125b-2-3p ( p  = 0.019) and − 200b-5p ( p  = 0.017). The multivariate model highlighted that high baseline miRNA-7a2-3p ( p  = 0.025), -21-5p ( p  = 0.014), -200b-3p ( p  = 0.041), young age ( p  = 0.042), lower bleeding on probing (BOP) ( p  = 0.021) and miRNA-125b-2-3p ( p  = 0.021) levels were significant predictors of OTM at 3-month follow-up. Conclusions In the present study, OTM significantly impacted the expression of the miRNAs analyzed, in both the tension and compression side of traction tooth at 3-month follow-up. High baseline miRNA-7a2-3p, -21-5p, -200b-3p, and lower miRNA-125b-2-3p, together with younger age and lower BOP, were significant predictors of OTM amount at 3-month follow-up. Trial registration ClinicalTrials.gov NCT06023433 (retrospectively registered).

Objective Nutraceutical agents have been demonstrated as adjuncts for the treatment of several inflammatory diseases. The present study analyzed and compared new nutraceutical agent as an adjunct to Scaling and root planing (SRP) versus SRP alone for the treatment of periodontitis. Materials and methods Sixty-six patients with moderate periodontitis were enrolled. Through a randomized design, the patients were randomly assigned to SRP + nutraceutical agent (test group) or SRP alone (control group). Patients were regularly examined the clinical, inflammatory mediators and visual analogue scale (VAS) changes over a 6-month period. Clinical attachment level (CAL) was the primary outcome variable chosen. Gingival crevicular fluid (GCF) inflammatory mediator change and the impact of treatment on VAS were evaluated through a linear regression model. Results Both treatments demonstrated an improvement in periodontal parameters compared with baseline. After 6 months of treatment, compared with the control group, the test group determined a significant probing depth (PD) ( p  = 0.003) and bleeding on probing (BOP) reduction ( p  < 0.001), while CAL gain was significantly obtained at 30 and 60 days after treatment ( p  < 0.05). In the test group, the level of inflammatory mediators was significantly reduced compared with the control group ( p  < 0.05). The linear regression analysis demonstrated that the nutraceutical agent exerted, in the test group, a significant influence on VAS at 6, 12, 24, and 48 h after treatment ( p  < 0.05). Conclusions Nutraceutical agent resulted in a more significant reduction in clinical, inflammatory mediators and short-term pain compared with SRP alone. Clinical relevance Nutraceutical agent, when combined with SRP, was demonstrated to be effective in reducing periodontal parameters and controlling the levels of inflammatory mediators and pain in patients with periodontitis.

Matrix metalloproteinase-9 (MMP-9) has been shown to play a key role in endothelial function and perhaps pivotal in the correlation between periodontal disease and cardiovascular disease (CVD). For the study, the impact of MMP-9 of periodontitis and CVD on serum and saliva concentrations was analyzed. For the study patients with periodontitis (n = 31), CVD (n = 31), periodontitis + CVD (n = 31), and healthy patients (n = 31) were enrolled. Clinical and demographic characteristics as well as serum and salivary MMP-9 were evaluated. MMP-9 concentrations in serum and saliva were statistically elevated in patients with CVD (p < 0.01) and in patients with periodontitis plus CVD (p < 0.001) compared to patients with periodontitis and healthy subjects. Multivariate regression analysis showed that c-reactive protein (hs-CRP) was the only significant predictor for MMP-9 serum (p < 0.001), whereas hs-CRP (p < 0.001) and total cholesterol (p = 0.029) were the statistically significant salivary MMP-9 predictors. This study evidenced that patients with CVD and periodontitis + CVD presented elevated MMP-9 concentrations in serum and saliva compared to patients with periodontitis and healthy subjects. Furthermore, hs-CRP was a negative predictor of serum and salivary MMP-9.

Background The aim of this study was to assess the association between serum and salivary Immunoglobulin (Ig) Aggregatibacter actinomycetemcomitans ( A. actinomycetemcomitans) specific antibodies in healthy controls (HC) and periodontitis (PT) patients. Furthermore, the objectives were to determine whether PT influenced serum A. actinomycetemcomitans specific antibodies and whether serum or salivary antibodies against A. actinomycetemcomitans IgG were mediated by serum high-sensitivity c-reactive protein (hs-CRP). Methods Fifty-three patients with periodontitis and 48 HC were enrolled in the present study. Patients were regularly examined and characterized by clinical, salivary and blood samples analyses. A. actinomycetemcomitans IgA and IgG antibodies and hs-CRP were evaluated using a commercially available kit. The Spearman Correlation Test and Jonckheere-Terpstra Test were applied in order to assess the interdependence between serum A. actinomycetemcomitans IgG antibodies and clinical periodontal parameters. To evaluate the dependence of the serum and salivary A. actinomycetemcomitans IgG levels from possible confounders, univariate and multivariable linear regression analyses were performed. Results Compared to HC, patients with PT had significantly higher IgA [serum: PT, 1.89 (1.2–2.2) EU vs HC, 1.37 (0.9–1.8) EU ( p  = 0.022); saliva: PT, 1.67 (1.4–2.1) EU vs HC, 1.42 (0.9–1.6) EU ( p  = 0.019)] and A. actinomycetemcomitans IgG levels [serum: PT, 2.96 (2.1–3.7) EU vs HC, 2.18 (1.8–2.1) EU ( p  < 0.001); saliva, PT, 2.19 (1.8–2.5) EU vs HC, 1.84 (1.4–2) EU ( p  = 0.028)]. In PT patients, serum A. actinomycetemcomitans IgG were associated with a proportional extent of PT and tooth loss (P-trend value< 0.001). The univariate regression analysis demonstrated that PT ( p  = 0.013) and high hs-CRP ( p  < 0.001) had a significant negative effect on serum and salivary A. actinomycetemcomitans IgG levels. The multivariate regression analysis showed that PT ( p  = 0.033), hs-CRP ( p  = 0.014) and BMI ( p  = 0.017) were significant negative predictors of serum A. actinomycetemcomitans IgG while hs-CRP (p < 0.001) and BMI ( P  = 0.025) were significant negative predictors of salivary A. actinomycetemcomitans IgG. Conclusions PT patients presented a significantly higher serum and salivary A. actinomycetemcomitans IgA and IgG compared to HC. There was a significant increase in serum A. actinomycetemcomitans IgG when patients presented a progressive extent of PT. Moreover, PT and hs-CRP were significant negative predictors of increased salivary and serum A. actinomycetemcomitans IgG levels. Trial registration The study was retrospectively registered at clinicaltrials.gov ( NCT04417322 ).

The aim of the present study was to investigate the association and impact of periodontitis and tooth loss on a subtype of endothelial progenitor cell (EPC) levels (CD133+/KDR+). Furthermore, the objective was to determine if the periodontal status influenced CD133+/KDR+ levels. In all, 88 patients with periodontitis and 79 healthy controls (HCs) were enrolled in the study. Enrolled patients were examined and characterized by clinical and blood sample analysis. Spearman’s correlation test was applied in order to assess the interdependence between CD133+/KDR+ levels and all periodontal parameters. In order to estimate a statistically significant trend (p-trend) for ordered CD133++/KDR+ quartiles, the Jonckheere–Terpstra test was applied for all variables. Patients in the periodontitis group presented significantly lower CD133+/KDR+ levels (66.4 (45.5–269.6 cells/µL)) compared to the HC group (76.7 (24.3–313.2 cells/µL), p < 0.001). Lower CD133+/KDR+ levels negatively correlated with C-reactive protein (CRP), with the number of teeth, and with all periodontal parameters (p < 0.001). Moreover, there was a proportional increase in CD133+/KDR+ levels with a progressive increase in number of teeth (p-trend < 0.001), while there was a proportional decrease in CD133+/KDR+ levels with a proportional increase in clinical attachment level (CAL, p-trend = 0.003), probing depth (PD, p-trend = 0.007), and bleeding sites (bleeding on probing (BOP), p-trend < 0.001) as an extent measure of periodontitis. This study demonstrated that patients with periodontitis presented significantly lower CD133+/KDR+ levels compared to HCs. Moreover, all patients presented an increase in the CD133+/KDR+ EPC levels with an extended level of periodontitis and tooth loss.

ObjectiveChemotherapeutic agents have been widely used as adjuncts for the treatment of chronic periodontitis (CP). This study investigated and compared a desiccant agent as an adjunct to scaling and root planing (SRP) versus SRP alone for the treatment of CP.Materials and methodsThirty-six patients with CP were studied. Using a split-mouth design, the maxillary right and left quadrants were randomly assigned to SRP plus desiccant (Hybenx® EPIEN Medical, Inc. St. Paul, MN, USA) or SRP alone. Patients were examined on a regular basis for clinical, microbiological, and inflammatory mediator changes over a 1-year period. Clinical attachment level (CAL) was the primary outcome variable. In addition, the red complex bacteria and gingival crevicular fluid (GCF) inflammatory mediators were monitored.ResultsCompared to baseline, both treatments demonstrated an improvement in periodontal parameters. Compared to SRP alone, SRP plus desiccant yielded a significant improvement in probing depth (PD) (SRP: 2.23 ± 0.31 mm vs. desiccant: 3.25 ± 0.57 mm, p < 0.05), CAL (SRP: 3.16 ± 0.29 mm vs. desiccant: 4.21 ± 0.34 mm, p < 0.05 mm) and bleeding on probing (BOP) (SRP: 4.56 ± 1.5% vs. desiccant: 34.23 ± 4.2%, p < 0.001) at 12 months. Similarly, in the SRP plus desiccant group, the bacteria of the red complex were significantly reduced (p < 0.05); and the level of inflammatory mediators was significantly reduced (p < 0.003) compared to SRP alone.ConclusionsSRP plus the desiccant resulted in a greater reduction in clinical, microbial and inflammatory mediators compared to SRP alone.Clinical relevanceDesiccant, when combined to SRP, was demonstrated as a significant approach to control the levels of certain periodontal pathogens, inflammatory mediators in patients with CP.

Background: The aim of this study was to compare the therapeutic effect of intravitreal treatment with ranibizumab and dexamethasone using specific swept-source optical coherence tomography retinal biomarkers in patients with diabetic macular edema (DME). Methods: 156 treatment-naïve patients with DME were divided in two groups: 75 patients received 3 monthly intravitreal injections of ranibizumab 0.5 mg (Lucentis®) (Group 1) and 81 patients received an intravitreal implant of dexamethasone 0.7 mg (Ozurdex®) (Group 2). Patients were evaluated at baseline (V1), at three months post-treatment in Group 1, and at two months post-treatment in Group 2 (V2). Best-corrected visual acuity (BCVA) and swept source-OCT were recorded at each interval. Changes between V1 and V2 were analyzed using the Wilcoxon test and differences between the two groups of treatment were assessed using the Mann–Whitney test. Multiple regression analysis was performed to evaluate the possible OCT biomarker (CRT, ICR, CT, SND, HRS) as predictive factors for final visual acuity improvement. Results: In both groups, BCVA improved (p-value < 0.0001), and a significant reduction in central retinal thickness, intra-retinal cysts, red dots, hyper-reflective spots (HRS), and serous detachment of neuro-epithelium (SDN) was observed. A superiority of dexamethasone over ranibizumab in reducing the SDN height (p-value = 0.03) and HRS (p-value = 0.01) was documented. Conclusions: Ranibizumab and dexamethasone are effective in the treatment of DME, as demonstrated by functional improvement and morphological biomarker change. DME associated with SDN and HRS represents a specific inflammatory pattern for which dexamethasone appears to be more effective.

Work organization, such as shifts and night work, can interfere with the perception of work-related stress and therefore on the development of pathological conditions. Night shift work, particularly, can have a negative impact on workers’ wellbeing by interfering with the biological sphere. The aim of this study is to evaluate the associations between work activities, shift work effects and stress-related responses in 106 dock workers enrolled in southeast Italy. Dock workers’ tasks consist of complex activities that seemed to affect more sleep quality than work-related stress. An analysis of salivary biomarkers such as cortisol, α-amylase, melatonin and lysozyme was performed along with validated psycho-diagnostic questionnaires. Alpha-amylase showed a significant negative correlation with the effort/reward imbalance score; thus, the measurement of salivary α-amylase is proposed as a sensitive and non-invasive biomarker of work-related stress. This study may provide new insights into developing strategies for the management of night shift work. Salivary biomarkers should be further investigated in the future in order to develop simple and effective tools for the early diagnosis of work-related stress or its outcomes.

Background/Objectives: Nickel–titanium (Ni-Ti) rotary instruments have significantly enhanced the efficiency and safety of root canal shaping. However, the variety of Ni-Ti systems, characterized by differences in alloy composition, taper design, and heat treatment, may influence their usability, particularly for novice operators. This study aimed to evaluate the perceptions of dental students using four Ni-Ti rotary systems (MTwo, SlimShaper Pro, ProTaper Gold, and HyFlex EDM) in a simulated environment. Methods: Forty dental students from two universities participated in the study and completed a standardized preoperative training session. Root canal shaping was performed on resin teeth models, and an 11-item questionnaire was used to assess various parameters, including flexibility, ease of use, and the ability to maintain working length. Results: The results indicated significant differences in student feedback. SlimShaper Pro and HyFlex EDM were preferred for their flexibility, reduced resistance, and ease of instrument transitions, likely due to their smaller tapers and martensitic alloys. MTwo was rated the least flexible and more challenging to use. Conclusions: These findings underline the importance of user-friendly Ni-Ti systems in pre-clinical training to reduce procedural difficulties and operator stress. Further research should validate these findings in clinical settings to improve the training of novice operators.