The use of casein in sport mouthguards: microbiological and ecological variations in oral cavity.

Sport mouthguards have the potential to become a microbial reservoir, produce oral and systemic diseases and cause negative changes in the oral cavity. The aim of this study was to monitor oral environmental changes caused by casein and sport-mouthguard in vivo, through clinical, salivary and bacterial markers of young athletes. Forty-eight active young athletes in different disciplines were selected and analysed at different times: baseline (T0); after three months of casein application on the mouthguard (T1); and after six months of application (T2). The product used was GC Tooth Mousse®. At T0, clinical monitoring was performed and the following parameters were recorded: Decay-Missing-Filled Teeth (DMFT) index, Plaque index (PL+) and Gingival Bleeding (BOP+). Saliva-Check Buffer GC® and Saliva-CheckMutans GC® salivary tests were then performed. At T0 the athletes demonstrated DMFT 0.03±0.01. PL value was positive in 100% of subjects at T0, T1, and T2. The BI value was always negative. At the three time-points, a significant change in baseline hydration values was observed; baseline viscosity was normal in 50% of cases while it increased in the remaining 50% at T0; it was normal and constant at T1 and T2. The value of the baseline pH underwent an not statistically significant increase at T1 (7.6±0.08) while remaining constant at T2. The amount of saliva produced after 5-min stimulation ranged significantly and gradually from T0 to T1 and T2, with a statistically significant difference. Plaque indicator tests highlighted that at T0 a plaque with a pH of 6.0±0.5 prevailed; at T1 it was 6.25±0.75 while at T2, pH was equal to 6. Tests for the detection of S. mutans resulted constant in all subjects at the various observation times, resulting in 67% of patients in whom S. mutans was present. The application of casein, within custom-made ethylene- vinyl acetate (EVA) mouthguards, positively influences salivary flow, the increase of pH values, the amount of stimulated saliva and the buffering capacity of the athlete, improving their state of oral health, which is negatively affected by the use of common mouthguards.

Mediterranean diet and physical activity improve posture, fat mass and salivary pH.

Many researchers have revealed that diet and physical activity influence metabolic function and posture in various stages of life. This paper aims to combine them and demonstrate how they could promote a healthy lifestyle. For this purpose, 14 healthy subjects followed a three-month protocol combining physical activity with dietary advice. At the end of the protocol, the results of the study underlined a significant reduction in fat mass, an improvement in salivary pH, and a realignment and rebalancing of body segments. .

The use of chlorhexidine in mouthguards.

Sports mouthguards have the potential to become a microbial reservoir, to produce oral and systemic diseases and cause changes in environmental oral factors, inhibiting the protective effect of saliva. The aim of this study was to monitor, in vivo, oral environmental changes caused by chlorhexidine and sports mouthguards and in vitro, the effects of microbial strains, saliva and chlorhexidine on Ethylene-Vinyl-Acetate (EVA) material. Seventy-two athletes were analyzed at different time points: before training session (T0), post-training (TA), post-training with mouthguard (TB), post-training with mouthguard and chlorhexidine (TC). At each time of observation, saliva was collected and subjected to microbiological analysis. In vitro, EVA disks were incubated with bacterial cultures, saliva and clorexidine. Culture of supernatant solution, SEM and bacterial counts of EVA disks were performed. S. mutans and Candida spp. load decreased significantly in TC. The pH value significantly decreased in TB and improved in TC. In vitro, the analyzed bacteria were organized to form a biofilm on the EVA disk surface. The addition of chlorhexidine to the bacterial culture and saliva inhibited the growth in all tested conditions. In vivo, the use of chlorhexidine associated with the sports mouthguard inhibited the growth of pathogenic microbial species, and improved pH values. In vitro, EVA stimulated biofilm formation on its surface, but this action was contrasted by chlorhexidine. The effects found in vitro encouraged the use of chlorhexidine in vivo as a valuable tool in the use of mouthguards.

Comparative molecular analysis of bacterial species associated with periodontal disease.

Periodontal disease is an inflammatory disorder affecting the supporting teeth structures, including gingiva, periodontal ligament and alveolar bone, causing loss of connective tissue, reabsorption of alveolar bone and formation of periodontal pockets. The aim of this study is to find a correlation between bacterial growth and periodontal disease. Fifty-seven patients aged between 21 and 65 years, median age 46 years, were enrolled. According to gingival pocket depth, ranging from 3 to 7 mm, patients were divided into two groups: the first (30 patients, 53%) with deep pockets ³ 5 mm and the second (27 patients, 47%) less than 5 mm. The samples taken were processed for microbiological analysis by absolute quantitative real-time Taq-Man technique. Patients affected by periodontal disease were 32 (56%) and patients with gingival bleeding were 35 (61%). This data showed that the presence, the type and the bacterial load in gingival pockets were strongly correlated with gingival depth, periodontal disease and gingival bleeding. Quantitative microbiological analysis is a key point to improve patient compliance, allowing to choose the specific antibiotic treatment. avoiding antibiotic resistance and ensuring the successful outcome of therapy for periodontal disease.

Black Stains: a microbiological analysis and a view on familiarity and susceptibility to tooth decay of patients in childhood.

AIM: Assess prevalence, familial predisposition and susceptibility to caries of Black Stains (BS). Evaluate the microbiological composition of BS, saliva and subgingival plaque. MATERIALS AND METHODS: Sixty nine subjects with BS (test group) and 120 subjects without BS (control group) were analysed for oral status. For each BS-patient, a BS-deposit, 1 ml of saliva and subgingival plaque were collected and microbiologically analysed. Five deciduous teeth with BS were observed under SEM. RESULTS: This study showed a BS prevalence similar to that of the Mediterranean area and a familiality. The microbiological origin of BS was confirmed by SEM and culture method and the BS flora differ from that of supragingival plaque. CONCLUSIONS: Predominance in BS and saliva of Actinomycetes and the low salivary prevalence of S. mutans and L. acidophilus may be related with low caries incidence in BS patients. The high presence of Actinomyces spp can be a causative factor for BS.

Hyaluronic acid-based medical device and oral disorders: can it be used in paediatric dentistry?

Due to its physical and biological characteristics and safety profile, hyaluronic acid is very widely used in numerous clinical conditions, ranging from its best-known use in cosmetic surgery (as a filler and for its ability to promote tissue regeneration and therefore minimise scarring) to lesser-known fields such as ophthalmic surgery, major abdominal surgery (where it is used to prevent the complication of adhesion bands) and intra-articular use. Studies were recently published in which this type of device was also used in paediatric patients for the management of inflammatory disorders of the oral cavity and teething symptoms. As this is a highly topical field for dentists, we felt it would be useful to review the efficacy and safety of the device in the paediatric population treated, and analyse any discrepancies with the results obtained in the adult population. The preparations of hyaluronic acid used in pediatric dentistry, thanks to their anti-inflammatory and angiogenic properties, proved to be very effective in therapy of oral diseases in children. Further clinical research is needed to confirm the effectiveness of these products to dispel doubts about any side effects.

Comparison of salivary antioxidant enzyme activity between ex-smokers and subjects who had never smoked.

Smoke contains oxidants such as oxygen-free radicals which are probably the major cause of damage to biomolecules. A decrease of salivary antioxidant enzymes was detected in habitual smokers. However, the effects of cigarette smoke on salivary antioxidant enzymes may persist after withdrawal from smoking. The objective of this study was to assess salivary superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activity in ex-smokers in comparison with that of subjects who had never smoked. The test group included 25 ex-smokers (13 males and 12 females; mean age: 48 ± 8 years) who had given up smoking for at least one year but for no more than 2 years, and a control group consisting of 25 subjects (14 males and 11 females; mean age: 50 ± 12 years) who had never smoked. Salivary samples were collected and SOD and GSH-Px activity was measured. Student’s t-test was used to evaluate differences between groups and significant differences were observed for p < 0.05. A significant decrease (p < 0.05) of GSH-Px (14.5 ± 2) was observed in the test group compared to the control group (30 ± 4). However, SOD was very similar in the two groups: 0.9 ± 0.3 in the test group and 0.8 ± 0.3 in the controls and no significant difference was detected (p> 0.05). Detoxification of hydrogen peroxide by the GSHPx was altered even after withdrawal from smoking, while the production of hydrogen peroxide, that is mediated by SOD, was not modified.

Effects of anti-epileptic drugs on the oral health of paediatric patients.

AIM: To evaluate the effects of epilepsy therapy on the oral health in paediatrics patients. MATERIALS: The test has involved 57 patients. The patients were stratified in three groups: monotherapy group, politherapy group and control group. They were examined and after that the test groups were compared with the control group. CONCLUSION: Paediatric epileptic patients seem to have a greater risk of having a worse oral health status compared with healthy patients. However, if the patient is well monitored and undergoes regular dental checks, the oral condition is comparable to a healthy subject.

Nitrous oxide and hypnotherapy in paediatric dentistry.

AIM: Patient compliance is of paramount importance for proper dental treatment, which can be difficult to achieve in children due to anxiety and fear, hindering the therapeutic procedure. In addition to well-known sedation methods (pharmacological and nitrous oxide), further methods of approach are being tested, such as hypnotherapy. MATERIALS: The sample consisted of 150 children (90 males, 60 females). Paper questionnaires, frequency meter, nitrous oxide dispenser in association with the use of the television were exploited. At the end of treatment, subjective pain was assessed using the VAS scale, while objective pain was assessed using the FLACC scale and heart rate. This made it possible to compare two experimental groups, one treated with nitrous oxide, and the other with hypnotherapy. The control group was treated with classic behavioural approach techniques. The results obtained were analysed by SPSS Software (Statistical Package for Social Science, version 27) and the inferences between the results were calculated for confirmation. CONCLUSION: Hypnotherapy can be a valid substitute for nitrous oxide in paediatric dentistry.

Enterococcus faecalis leakage of root canal sealers: an ex vivo study.

The aim of this ex vivo study was to evaluate bacterial penetration after filling root canals using 3 different techniques. Three experimental groups of 25 teeth each, obturated with lateral-warm-vertical condensation of gutta-percha, Microseal technique and EndoREZ system, respectively, were tested in a split chamber model system using Enterococcus faecalis and monitored for 180 days to determine bacterial penetration. A statistical analysis was performed using the Kaplan-Meier method. Median survival time was 25 days for Microseal system, 41 for lateral-warm-vertical condensation and 81 for EndoREZ®. Significant differences were demonstrated between Microseal and EndoREZ (p less than 0.001) and between Microseal and lateral-warm-vertical condensation technique (p less than 0.05). No statistically significant differences were observed between EndoREZ and lateral-warm-vertical condensation. After 180 days of assessment, 20 percent of the EndoREZ samples resisted bacterial penetration and furthermore, the EndoREZ system has the potential to be a filler system compatible with other currently used systems.

Dental and periodontal complications of labial and tongue piercing.

Piercing is the practice of puncturing some parts of the body to apply ornamental objects. The presence of oral and perioral piercings are a risk factor for many acute and chronic complications, such as chipping of the dental enamel, periodontal lesions and infection. The aim of this study is to assess the prevalence of lip and tongue piercing complications in the dental and periodontal tissues in a sample of young adults. Twenty-five adult patients were examined (test group: 11 males and 14 females with an average age of 23.4+/-3.6 years) who had had a minimum of one labial or tongue piercing for at least 1 year and were compared with 25 subjects (control group: 11 males - 44 percent, and 14 females - 56 percent) without any lingual or labial piercing. A questionnaire was compiled for each patient and a clinical examination was performed. The following parameters were examined by the same operator: abnormal toothwear, tooth chipping or cracking, clinical attachment loss (CAL), probing pocket depth (PD) and gingival recession (GR, classified by using Miller s classification). The data were analyzed using X2 or Fisher s exact test for small numbers and non-parametric Mann-Whitney or Kruskal-Wallis tests to examine for differences in continuity; the level of significance was p less than 0.05. According to the results found in the present study the prevalence of abnormal tooth wear and tooth chipping was higher in the subjects with labial or lingual piercing. Moreover, patients with tongue or labial piercing exhibited a higher GR in comparison to control subjects without any oral piercing. No differences were observed between the two groups as regards CAL and PD. A significant association between the duration of piercing and dental defects was found in the group of patients with piercings with greater prevalence of tooth and periodontal defects in the group of 13 subjects who had had the piercings for a period less than or equal to 4 years.

Management of compound odontoma in a 10-year-old girl preserving the associated impacted permanent tooth.

AIM: Compound odontoma has been reported to be the most common of all odontogenic neoplasms and tumor- like lesions. Only rarely the treatment of this lesion in association with an impacted tooth has been reported. CASE REPORT: A compound odontoma in a 10-year-old girl, associated with an impacted permanent incisor is described, focusing on the diagnosis and the importance of early treatment of this lesion. The patient underwent surgical excision of the lesion and it was decided to wait for the spontaneous eruption of the impacted tooth. After 6 months no eruption was observed and thus the orthodontic treatment was deemed necessary. At the one-year follow-up, the tooth was brought into the maxillary arch.

Clowntherapy in paediatric dentistry: The psychological approach to the paediatric patient.

AIM: Anxiety towards dental treatments and visits in children are often cause for the avoidance and/or failure of the treatment itself, as well as a reason for stress in both the patient and the orthodontist. The aim of the research is to understand if the use of clown therapy in dental waiting rooms is efficient in bringing down the level of preoperative anxiety in paediatric dental patients. METHODS: Two psychometric tests were used: CFSS-DS (Dental Subscale of the Children's Fear Survey Schedule) and FIS (Facial Image Scale), after the translation of the original versions, to both the patients and their parents. The CFSS-DS is the most used instrument in order to assess dental fear in children, it is composed by 15 questions that outline situations which children and adolescents will have to face while on the dental chair. The FIS is a visual analogical scale that uses faces as indicators of anxiety. CONCLUSION: Clowntherapy proves to be an efficient non-pharmacological method to reduce anxiety and dental fear in paediatric patients, offeringa valid and practical support for paediatric dentists.

Effect of 2-hydroxyethyl methacrylate on Streptococcus spp. biofilms.

AIMS: The effect of different concentrations of 2-hydroxyethyl methacrylate (HEMA) was evaluated on biofilm formation and preformed biofilm of Streptococcus mitis, Streptococcus mutans and Streptococcus oralis, alone or combined to each other. METHODS AND RESULTS: Twofold serial dilution of HEMA ranged from 12 to 0·75 mmol l(-1) was added to Streptococcal broth cultures and mature biofilms in 96-well-microtitre plates to evaluate bacterial biomass and cell viability. HEMA affected the Streptococcal population in a strain-specific way producing few significant effects. A reduction on biofilm formation and a detachment of preformed biofilm was recorded in Strep. mitis ATCC 6249, whereas in mixed cultures, the monomer expressed a general aggregative effect on mature biofilms. A reduction in cell viability was also recorded in an HEMA-concentration-dependent way in each experimental condition studied. CONCLUSIONS: These results suggest that the HEMA prevalent effects are both the reduction of bacterial adhesion to a polystyrene surface and the increase in dead cells also characterized by an aggregative status. SIGNIFICANCE AND IMPACT OF THE STUDY: Understanding the potential effect of HEMA, released from resin-based materials, on oral bacteria may furnish information for surveillance of the risk reduction in secondary caries via hindering biofilm generation.

Anti-adhesive and pro-apoptotic effects of 2-hydroxyethyl methacrylate on human gingival fibroblasts co-cultured with Streptococcus mitis strains.

AIM: To evaluate and observe the cellular reactions that occur during the interaction/integration between 2-hydroxyethyl methacrylate/host tissue/microbial environment, in a co-culture of human gingival fibroblasts (HGF) and Streptococcus mitis strains. METHODOLOGY: Streptococcus mitis were cultured with strains in the presence of 3 mmol L(-1) HEMA for 48 h and 72 h. Cytotoxicity was evaluated by the trypan blue dye exclusion test. Apoptosis was evaluated by TUNEL analysis. Adhesion was evaluated by immunofluorescence and western blot analyses. Quantitative analyses of the results were acquired by Qwin Plus 3.5 and QuantityOne I-D analysis software, respectively. The statistical significance of the results was evaluated using t-tests and linear regression tests. RESULTS: The trypan blue dye test revealed 47.3% and 46.5% of dead fibroblasts after 48 and 72 h HEMA treatment, respectively, while bacterial viability was not influenced by the presence of HEMA and fibroblasts. The expression of pro-collagen I, involved in fibroblast adhesion, in untreated samples ranged from 12.49% to 6.91% of the positive area after 48 and 72 h, respectively, dropping to below 2% of the positive area in the other experimental conditions. Unlike the trypan blue test, co-cultured samples treated with HEMA showed 20% and 25% versus 17% and 21% (after 48 and 72 h, respectively) of apoptotic cells. CONCLUSIONS: The evidence for HEMA toxicity and anti-adhesive effects against eukaryotic cells was reduced in the presence of bacteria, suggesting that dental resins should be well polymerized to avoid the spread of toxic monomers within the mouth.

Wear of dental sealing materials using the replication technique.

AIM: The aim of the present study was an in vivo evaluation of the wear of different sealing materials using the replication technique. MATERIALS AND METHODS: Four different sealing materials were randomly applied on 2 out of 4 caries-free permanent first molars of each patient. Impressions were taken at T0: before the sealing material was applied; T1: right after the application; T2: 6 months after the application, and T3: 1 year after the application. The impressions were then processed for scanning electron microscopy analysis of the wear of the examined materials. Statistical analysis was used to evaluate differences among and within the 4 groups. RESULTS: At T2, the average ratio of residual material in all the groups was 62.37% ± 1.21%, while at T3 it was 49.63% ± 1.11%. At both T2 and T3 there were no statistically significant differences among the 4 groups examined (p=0.76 and p=0.50 respectively). Comparison within the same group at T2 and T3 showed that statistically significant differences were only detected in group D (p=0.015). CONCLUSION: Sealing materials do not undergo a complete loss after 1 year of application, thus indicating that they are suitable for caries prevention.

The use of customized mouthguards during the training produced protective effects on salivary factors of young athletes.

AIM: Custom-made mouthguards have many advantages compared to the stock and ready-made types, but sport treatments with custom made mouthguards involve changes in ecological factors of the oral cavity. In the present study we investigated the potential protective role of salivary factors, such as pH value, volume, prostaglandin E2 (PGE2) and 8-iso-prostaglandin F2? (8-iso-PGF2?) levels during training with customised mouthguards. MATERIALS AND METHODS: A total of 80 subjects were selected: 40 athletes, of whom 20 practice volleyball and 20 basketball (test group), and 40 subjects who attend a gym at a non-competitive level (control group). The athletes (test group) were analyzed at baseline (T0), pre-training (T1), post-training with custom-made Ethylene-Vinyl-Acetate (EVA) mouthguards (T2), post-training without mouthguards (T3). The control group was analyzed only at baseline (T0). On each player, in the 4 time points, and on the control group at T0, we stimulated saliva for determining PGE2 and 8-iso-PGF2? levels by radioimmunoassay and pH value by a pH meter and volume/ml. Saliva pH was calculated with a pH meter. RESULTS: We observed an inhibition of 8-iso-PGF2? salivary release induced by physical exercise and by use of custom-made mouthguard, while we found an increase in PGE2 salivary level in athletes after training and wearing the mouthguard. Furthermore, in the test of the volume of saliva produced in 5 minutes, a significant inhibition of saliva production emerged in the athletes who did not use the mouthguard during sports activities. CONCLUSION: Sports activity could lead to a reduction in oxidative stress and the use of mouth guards seems even more effective for athletes.

Short lingual frenulum: From diagnosis to laser and speech-language therapy.

AIM: To evaluate the effects of tongue frenulectomy performed with two therapeutic approaches: Laser frenulectomy and combined laser and speech-language therapy. MATERIALS AND METHODS: The study involved 180 patients (90 males and 90 females) aged between 6 and 12 years. After examination and data collection, the patients were stratified according to three degrees of severity: mild, moderate and severe. After treatment, the test group (laser frenulectomy and combined laser and speech-language therapy) was compared with the control group (laser frenulectomy) in the pre-surgical phase, at one week, 1 month, 3 months, 6 months and 12 months after surgery. RESULTS: Statistical analysis showed statistically significant differences between the pre-surgical and post-surgical values at 3 months, 6 months and 12 months after surgery (p<0,05). CONCLUSIONS: It is essential to establish diagnosis criteria to which the clinician should refer in order to decide the treatment plan. This study shows that combined laser and speech-language therapy leads to better results than the resection treatment of the frenulum with laser technique alone.

Effects of cigarette smoke on salivary superoxide dismutase and glutathione peroxidase activity.

Cigarette smoke contains oxidants such as oxygen-free radicals and volatile aldehydes, which are probably the major causes of damage to biomolecules exposed to cigarette smoke. However, saliva has an antioxidant defense system able to counter toxic activities of radical species that is formed by antioxidant enzymes, including superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px). The purpose of this study is to verify the possible effects of cigarette smoke on SOD and GSH-Px. Forty-four patients (25 males and 19 females) were enrolled in this study. The participants were 20 smokers (12 males and 8 females) and 24 non-smokers (13 males and 11 females). Furthermore, 10 subjects of the control group were ex-smokers (9 males and 1 female). Their mean age plus or minus standard deviation (SD) was 58.8 plus or minus 15.9 years for the case group and 73.8 plus or minus 10.6 years for the control group. All patients were underwent a careful anamnestic investigation and examination of the oral cavity. After rinsing the mouth with water, each subject put 3 cc of non-stimulated saliva inside a test tube. The saliva was centrifuged and oral peroxidase and superoxide dismutase activity was measured according to a specific assay. Statistical analysis was performed to evaluate differences between the groups and significant differences were observed for p less than 0.05. A significant decrease of GSH-Px activity was detected in the smoking group (p less than 0.05), while the SOD activity was similar in the control and case groups. According to the sex, a significant decrease of GSH-Px activity was noted in males of the smoker group (p less than 0.05), while in the sample of females no significant difference of the enzymatic activity was found. Moreover, among ex-smokers, there was a significant difference in the values of GSH-Px between those who had not smoked for less than ten years and those who had not smoked for more than ten years. Cigarette smoke may alter the detoxification of hydrogen peroxide through a decrease of GSH-Px activity. The overproduction of H2O2 may lead to an oxidative stress that is involved in a large number of diseases, including precancerous and neoplastic lesions of the oral cavity. The effects of cigarette smoke on salivary antioxidant enzymes decrease after withdrawal from smoking and the benefits become more evident with the passage of time.

Effects of a novel gel containing 5-aminolevulinic acid and red LED against bacteria involved in peri-implantitis and other oral infections.

Antibiotic resistance is a major public health problem worldwide and the finding of alternative methods for eliminating bacteria is one of the prerogatives of medical research. The indiscriminate use of antibiotics in dentistry, especially for the treatment of peri-implantitis, could lead to superinfections. Alternative methods, like photodynamic therapy mediated by the use of aminolevulinic acid and a red light has been largely described, especially in dentistry, but results were encouraging against Gram-positive bacteria, but limited against Gram-negative. The effectiveness of photodynamic therapy mediated by a novel product containing aminolevulinic acid, Aladent (ALAD) has been tested in this in vitro study, against different types of bacteria particularly involved in the infections of the oral cavity and peri-implantitis. The novelty of ALAD is the marked hydrophilicity that should increase the passage of the molecule through the membrane pores of Gram-negative bacteria. Considering the novelty of the product a preliminary experiment permitted to test the effectiveness against Enterococcus faecalis after 1 h of ALAD incubation at different concentrations, with or without different timings of LED irradiation. The count of CFUs and the live/dead observation with fluorescent microscopy showed a significant reduction and killing of bacterium. Then, in the second stage, that could meet the necessity of effectiveness and the clinician's requests to reduce the timing of treatment, ALAD, with and without irradiation, was tested on Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Veillonella parvula and Porphyromonas gingivalis. In particular, the efficiency of different concentrations of the product after a 25 min incubation was tested with and without the adjunctive LED irradiation for 5 min. A slight ALAD bactericidal effect was reported for all bacteria, also without LED irradiation, however, the most effective treatment was 25 min of 50% ALAD incubation followed by 5 min of a red LED. The in vitro tests demonstrated that ALAD gel with LED irradiation exerts a potent antibacterial activity on different bacteria, both Gram-positive and Gram-negative.