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BACKGROUND AND AIMS: Apolipoprotein B plays a crucial role in regulating plasma cholesterol by mediating the interaction of low-density lipoprotein (LDL) with LDL receptors in the liver. Inherited mutations in this gene may increase the risk of developing premature atherosclerotic cardiovascular disease, especially in individuals with familial hypercholesterolemia type 2 (FH2). The aim of this study is to identify APOB variants that may indicate pathogenicity in a sample of the Brazilian population using a data bank exome sequencing study by NGS in a Brazilian population phenotypically diagnosed by clinical and laboratory profile. This finding is going to improve genetic hypercholesteremia diagnosis. METHODS: High-quality DNA samples (n»300) were sequenced using an exon-targeted gene sequencing (ETGS) strategy to identify variants in FHrelated genes. Pathogenicity classification was based on criteria established by the American College of Medical Genetics and Genomics (ACMG), also using information from ClinVar and pathogenicity scores from previous association studies. RESULTS: A total of 121 variants were identified in APOB, of which four are novel variants missense (p.Thr626Asn, p.Ile2750Thr, p.Gln2078Lys and p.Met4184Arg). After curating pathogenicity scores, variants were classified according to the ACMG criteria. Among them four as pathogenic or likely pathogenic (p.Pro2739Leu, p.His1923Arg, p.Pro994Leu and p.Pro877Leu), and 21 variants had uncertain significance. Additionally, 92 previously known variants with uncertain significance were classified as benign or likely benign. The results were submitted to Clinvar for actualization of pathogenicity. CONCLUSIONS: These results improve the molecular diagnosis associating APOB variants with the clinical phenotype of hypercholesterolemia.

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OBJECTIVE: Considering that misfit is a significant predictor of the clinical success of indirect restorations, the objective of this study was to evaluate the marginal and internal misfit of two computer-aided design and manufacturing (CAD/CAM) RMC ceramic materials used as occlusal veneers (OVs) of different thicknesses. METHODS AND MATERIALS: A CAD model of a mandibular first molar was obtained and OV preparations 0.5-, 1.0-, and 1.5-mm thick were modeled and milled in two different materials (n=10/group): resin nanoceramic (RNC) and polymer-infiltrated ceramic network (PICN). Using the same CAD model, tooth preparations were milled in fiber-reinforced epoxy resin (n=20/thickness). The marginal and internal misfit of the restorations was assessed by X-ray microtomography. The measurements of the marginal gap (MG) and absolute marginal discrepancy were performed in two locations on each slice, whereas internal gap (IG) measurements were performed at ten locations on each slice. The data obtained were analyzed using two-way analysis of variance and Tukey post-hoc tests (α=0.05). RESULTS: No significant effect was attributable to the material type or material-thickness interaction for the MG, absolute marginal discrepancy (AMD), or IG (p>0.05). However, the thickness significantly affected the IG of the restorations (p<0.05). CAD/CAM RNC and PICN systems presented similar MG and AMD for OVs 0.5-, 1.0-, and 1.5-mm thick. However, the IG varied between thicknesses.

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BACKGROUND: Carbapenem-resistant Pseudomonas aeruginosa (CRPA) infection after kidney transplantation (KT) is associated with high mortality. AIM: To analyse an outbreak of infection/colonization with IMP-1-producing CRPA on a KT ward. METHODS: A case-control study was conducted. Cases were identified through routine surveillance culture and real-time polymerase chain reaction for carbapenemase performed directly from rectal swab samples. Controls were randomly selected from patients hospitalized on the same ward during the same period, at a ratio of 3:1. Strain clonality was analysed through pulsed-field gel electrophoresis (PFGE), and whole-genome sequencing was performed for additional strain characterization. FINDINGS: CRPA was identified in 37 patients, in 51.4% through surveillance cultures and in 49.6% through clinical cultures. The median persistence of culture positivity was 42.5 days. Thirteen patients (35.1%) presented a total of 15 infections, of which seven (46.7%) were in the urinary tract; among those, 30-day mortality rate was 46.2%. PFGE analysis showed that all of the strains shared the same pulsotype. Multilocus sequence typing analysis identified the sequence type as ST446. Risk factors for CRPA acquisition were hospital stay >10 days, retransplantation, urological surgical reintervention after KT, use of carbapenem or ciprofloxacin in the last three months and low median lymphocyte count in the last three months. CONCLUSION: KT recipients remain colonized by CRPA for long periods and could be a source of nosocomial outbreaks. In addition, a high proportion of such patients develop infection. During an outbreak, urine culture should be added to the screening protocol for KT recipients.

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Serum thyroglobulin is used as part of the early postoperative assessment of differentiated thyroid cancer (DTC) since there is a clear relationship between an increased risk of recurrence and persistent disease after initial treatment and high postoperative stimulated thyroglobulin (ps-Tg) values. Thus, although ps-Tg above 10-30 ng/mL is considered an independent predictor of worse prognosis, the value that is associated with distant metastases is not defined. Thus, this was our objective. We selected 655 DTC patients from a nuclear medicine department database (Irmandade Santa Casa de Misericórdia de São Paulo, Brazil). All patients had received total thyroidectomy and radioactive iodine (RAI) therapy and had ps-Tg values higher than 10 ng/mL with negative anti-thyroglobulin antibodies. Then, we selected patients who presented post-therapy whole-body scan with pulmonary and/or bone uptake but with no mediastinum or cervical uptake. Patients with negative findings on functional imaging or any doubt on lung/bone uptake were submitted to additional exams to exclude another non-thyroid tumor. Of the 655 patients, 14.3% had pulmonary and 4.4% bone metastases. There was a significant difference in ps-Tg levels between patients with and without metastases (P<0.001). The cutoff value of ps-Tg was 117.5 ng/mL (sensitivity: 70.2%; specificity: 71.7%) for those with lung metastasis, and 150.5 ng/mL (sensitivity: 79.3%; specificity: 85%) for those with bone metastasis. The cutoff value for patients with eitherpulmonary or bone metastasis was 117.5 ng/mL (sensitivity: 70.2%; specificity: 83.7%). Our findings demonstrated that ps-Tg could predict distant metastasis in DTC patients. We identified a cutoff of 117.5 ng/mL with a high negative predictive value of 93.7%.

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Serum thyroglobulin is used as part of the early postoperative assessment of differentiated thyroid cancer (DTC) since there is a clear relationship between an increased risk of recurrence and persistent disease after initial treatment and high postoperative stimulated thyroglobulin (ps-Tg) values. Thus, although ps-Tg above 10-30 ng/mL is considered an independent predictor of worse prognosis, the value that is associated with distant metastases is not defined. Thus, this was our objective. We selected 655 DTC patients from a nuclear medicine department database (Irmandade Santa Casa de Misericórdia de São Paulo, Brazil). All patients had received total thyroidectomy and radioactive iodine (RAI) therapy and had ps-Tg values higher than 10 ng/mL with negative anti-thyroglobulin antibodies. Then, we selected patients who presented post-therapy whole-body scan with pulmonary and/or bone uptake but with no mediastinum or cervical uptake. Patients with negative findings on functional imaging or any doubt on lung/bone uptake were submitted to additional exams to exclude another non-thyroid tumor. Of the 655 patients, 14.3% had pulmonary and 4.4% bone metastases. There was a significant difference in ps-Tg levels between patients with and without metastases (P<0.001). The cutoff value of ps-Tg was 117.5 ng/mL (sensitivity: 70.2%; specificity: 71.7%) for those with lung metastasis, and 150.5 ng/mL (sensitivity: 79.3%; specificity: 85%) for those with bone metastasis. The cutoff value for patients with eitherpulmonary or bone metastasis was 117.5 ng/mL (sensitivity: 70.2%; specificity: 83.7%). Our findings demonstrated that ps-Tg could predict distant metastasis in DTC patients. We identified a cutoff of 117.5 ng/mL with a high negative predictive value of 93.7%.

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Botulinum toxins (BoNTs) are a product of the bacteria Clostridium botulinum. By entering nerve endings, they cleave and inactivate SNARE proteins, which are essential for neurotransmitter release. Prevention of acetylcholine release at the neuromuscular junction causes long-lasting and potentially fatal flaccid paralysis-a major feature of botulism. However, an intramuscular injection of minute amounts of BoNTs, primarily type A (BoNT-A), has useful long-lasting muscle relaxation effects on spastic motor disorders. This characteristic of BoNT-A is widely used in neurology and cosmetics. Over the last few decades, it has been demonstrated that the functions of BoNT-A are not limited to muscle-relaxing or autonomic cholinergic effects but that it can act as an analgesic agent as well. More recently, it was revealed that this antinociceptive effect starts after entering the sensory nerve endings, where these agents are axonally transported to the central nervous system, suggesting that at least part of their analgesic effect might be of central origin. Because of its antinociceptive effect, BoNT-A is currently approved for treatment of chronic migraine; nonetheless, case reports and preclinical and clinical experiments indicating its benefit in numerous potential painful conditions have increased. In the field of dentistry, the US Food and Drug Administration approved BoNT-A for the treatment of sialorrhea only. Legal status of the use of BoNT-A in other countries is less known. However, there are controlled clinical trials suggesting its efficacy in other conditions, such as bruxism, temporomandibular disorders, and trigeminal neuropathic pain. Thereby, using criteria of the American Academy of Neurology, we critically reviewed the uses of BoNTs in oral medicine and found it effective for trigeminal neuralgia (category A) and probably effective in temporomandibular disorders and bruxism.

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This study reviews the clinical and radiographic outcomes of Mini-implants (MI) and Narrow Diameter Implants (NDI) as mandibular overdenture (MO) retainers. Six databases were consulted for clinical studies that evaluated implants with diameter ≤3.5 mm. Data on the MI and NDI for survival and success rate and peri-implant bone loss and were collected and submitted to meta-analysis. Thirty-six studies were included, 24 reporting MI performance and 12 describing NDI results. The MI group comprised data from 1 cross-sectional clinical study, 3 retrospective longitudinal (RL) clinical studies, 13 prospective longitudinal (PL) clinical studies and 7 randomised clinical trials (RCT) with follow-up periods ranging from 1 day to 7 years. Eight studies used conventional loading, thirteen used immediate loading, two studies used both loading types, and one study did not report. The NDI group comprised data from 3 RL clinical studies, 6 PL clinical studies and 3 RCT with follow-up ranging from 6 months to 10 years. Ten studies used conventional loading, 1 study used immediate loading, and 1 study did not report. The average survival rates of MI and NDI studies were 98% and 98%, respectively, while the average success rates were 93% and 96%, respectively. The average peri-implant bone loss after 12, 24 and 36 months was 0.89, 1.18 and 1.02 mm for MI and 0.18, 0.12 and -0.32 mm for NDI. Both MI and NDI showed adequate clinical behaviour as overdenture retainers. The NDI showed a better long-term predictability to retain OM with most studies adopting conventional loading.

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BACKGROUND: Carbapenem-resistant Enterobacteriaceae (CRE) have been reported worldwide and are associated with high mortality rates. Intestinal colonization acts as a reservoir and fosters exchange of resistance mechanisms. AIM: To investigate the prevalence of patients harbouring CRE on hospital admission, risk factors associated, and the acquisition rate within the emergency department (ED). METHODS: This was a cross-sectional survey with 676 patients consecutively admitted to the ED study during the months of May to July 2016. A questionnaire was performed and rectal swabs were collected from patients on admission, for culture and for multiplex real-time polymerase chain reaction (PCR). If the patient was hospitalized for more than one week in the ED, samples were taken again to determine the acquisition rate of CRE. FINDINGS: Forty-six patients were colonized; all positive PCR were Klebsiella pneumoniae carbapenemase. The acquisition rate was 18%. Previous exposure to healthcare in the last year, liver disease, and use of antibiotics in the last month were risk factors for colonization. Six patients with no previous exposure to healthcare were CRE-colonized on admission, suggesting transmission of CRE within the community. CONCLUSION: Screening of high-risk patients on admission to the ED is a strategy to early identify CRE carriage and may contribute to control CRE dissemination.

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This longitudinal clinical study investigated the differences in the masticatory function (MF), satisfaction and oral health-related quality of life (OHRQoL) between atrophic patients (AP) and non-atrophic patients (NAP) before and after rehabilitation with mandibular overdenture (MO). Twenty-six complete denture (CD) wearers were categorised into two groups, according to the mandibular bone atrophy (MBA) degree. MF was evaluated before and after 1, 3, 6 and 12 months of the MO loading via 2 standardised tests: (i) MP, masticatory performance (MP_X50, MPB, ME 5·6, ME 2·8) and (ii) ST, swallowing threshold (time, number of cycles, ST_X50, STB, ME 5·6, ME 2·8). The dental impact on daily living (DIDL) questionnaire measured changes in the satisfaction level and OHRQoL. MP comparisons showed significant difference only for ME 5·6 12 months after MO loading (AP=33·79 ± 23·6; NAP=17·58 ± 20·1). ST presented significant differences before MO loading for: ST_X50 (AP=5·48 ± 0·83; NAP=4·31 ± 1·44), ME 5·6 (AP=53·17 ± 24·71; NAP=29·83 ± 31·45) and ME 2·8 (AP=8·76 ± 6·91; NAP=18·61 ± 10·71). One month after MO loading, NAP performed the ST test 21% faster than AP. After 3 months, significant improvements in STB (AP=4·93 ± 4·82; NAP=2·73 ± 1·27) and ME 2·8 (AP=17·15 ± 10·00; NAP=24·69 ± 7·82) also were observed. DIDL evaluation showed significant differences in the oral comfort domain after 3 months (AP=0·66 ± 0·29; NAP=0·87 ± 0·16) and after 6 months (AP=0·79 ± 0·22; NAP=0·98 ± 0·08), with lower satisfaction levels in the AP. MBA negatively affects the MF mainly the ST. After 6 months, differences between AP and NAP disappeared and ST results were equalised. AP initially has lower satisfaction levels reaching similar levels of satisfaction as NAP after 1 year.

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OBJECTIVES: The aim of this study was to investigate the restoration of broken-down endodontically treated incisors with the ferrule effect using glass ceramic crowns bonded to composite resin core buildups with or without a fiber post. A no-ferrule group with post was also included for comparison. METHODS AND MATERIALS: Thirty decoronated endodontically treated bovine incisors with a 2-mm ferrule were restored with a direct buildup using a nanohybrid direct composite resin (Miris 2 and Optibond FL) with or without a glass-fiber-reinforced post. An additional group of 15 teeth without a ferrule were restored with buildup and a fiber post. All teeth were prepared to receive bonded glass ceramic crowns (e.max CAD luted with Variolink Esthetic DC) and were subjected to accelerated fatigue testing. Cyclic isometric loading was applied to the incisal edge at an angle of 30° and a frequency of 5 Hz, beginning with a load of 100 N (×5000 cycles). A 100 N load increase was applied each 15,000 cycles. Specimens were loaded until failure or to a maximum of 1000 N (×140,000 cycles). Groups were compared using the Kaplan Meier survival analysis (log rank test at p=0.05). RESULTS: None of the tested specimens withstood all 140,000 load cycles. Specimens with posts but without a ferrule were affected by an initial failure phenomenon (wide gap at the lingual margin between the buildup/crown assembly and the root). There was a significant difference in mean survived cycles between the ferrule groups (Fp=73,332× and FNp=73,244×) and the no-ferrule group (50,121×; p=0.001). The addition of a fiber post was not significant in the presence of the ferrule (p=0.884). In both groups with posts, 100% of failures were unrestorable. The no-post group had 47% of restorable and possibly restorable failures. CONCLUSIONS: The survival of broken-down nonvital incisors was improved by the presence of the ferrule but not by the fiber-reinforced post. Fiber posts were always detrimental to the failure mode and were not able to compensate for the absence of a ferrule.

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The fungus Candida albicans is carried orally and causes a range of superficial infections that may become systemic. Oral bacteria Actinomyces oris and Streptococcus oralis are abundant in early dental plaque and on oral mucosa. The aims of this study were to determine the mechanisms by which S. oralis and A. oris interact with each other and with C. albicans in biofilm development. Spatial distribution of microorganisms was visualized by confocal laser scanning microscopy of biofilms labeled by differential fluorescence or by fluorescence in situ hybridization (FISH). Actinomyces oris and S. oralis formed robust dual-species biofilms, or three-species biofilms with C. albicans. The bacterial components tended to dominate the lower levels of the biofilms while C. albicans occupied the upper levels. Non-fimbriated A. oris was compromised in biofilm formation in the absence or presence of streptococci, but was incorporated into upper biofilm layers through binding to C. albicans. Biofilm growth and hyphal filament production by C. albicans was enhanced by S. oralis. It is suggested that the interkingdom biofilms are metabolically coordinated to house all three components, and this study demonstrates that adhesive interactions between them determine spatial distribution and biofilm architecture. The physical and chemical communication processes occurring in these communities potentially augment C. albicans persistence at multiple oral cavity sites.

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Alendronate therapy has been associated with serious side effects. Altering the alendronate concentration and combining with high-frequency loading as mechanical intervention was explored in this animal study as a treatment for osteoporosis. The bone anabolic potency of high-frequency loading was overruled by the different alendronate dosages applied in the present study. Further exploration of reduced hormonal therapy associated with mechanical interventions in osteoporosis treatment should be sought. INTRODUCTION: The aim of the present study was to investigate the effect of alendronate (ALN) administration at two different dosages, associated or not with high-frequency (HF) loading, on the bone microstructural response. METHODS: Sixty-four female Wistar rats were used, of which 48 were ovariectomized (OVX) and 16 were sham-operated (shOVX). The OVX animals were divided into three groups: two groups were treated with alendronate, at a dosage of 2 mg/kg (ALN(2)) or at a reduced dosage of 1 mg/kg (ALN(1)) three times per week. A third OVX group did not receive pharmaceutical treatment. All four groups were mechanically stimulated via whole body vibration (WBV) at HF (up to 150 Hz) or left untreated (shWBV). ALN and HF were administered for 6 weeks, starting at 10-week post-(sh)OVX. Tibia bone structural parameters were analyzed using ex vivo microcomputed tomography. RESULTS: Trabecular bone loss and structural deterioration resulting from ovariectomy were partially restored by ALN administration, demonstrated by the improvement of trabecular patter factor (Tb.Pf), trabecular separation (Tb.Sp), and structure model index (SMI) of the ALN groups compared to that of the OVX group, regardless of the applied dosage [ALN(2) or ALN(1)] or mechanical loading regime (shWBV or WBV). However, a significant positive effect of the ALN(1) administration on trabecular (decrease of Tb.Sp and SMI) and cortical bone (increase of cortical thickness) microarchitecture compared to that of the OVX status group was observed for both loading regimes was not seen for ALN(2). Furthermore, HF loading resulted in cortical bone changes, with an increased trabeculary area and endocortical perimeter. Finally, the benefits of a combined therapy of ALN with HF loading could not be discerned in the present experimental conditions. CONCLUSIONS: The bone anabolic potency of HF loading was overruled by the ALN dosages applied in the present study. Further altering the ALN dosage combined with robust mechanical stimuli needs to be considered in osteoporosis research and eventually therapy.

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Monolithic zirconia crowns have many favorable properties and may potentially be used to solve dental problems such as chipping. However, monolithic zirconia crown resistance can be affected by its phase transformation when subjected to low temperatures, humidity, and stress. This study evaluated the fracture load and phase transformation of monolithic zirconia crowns submitted to different thermal and mechanical aging tests. Seventy monolithic zirconia crowns were randomly divided into the following five groups: control, no treatment; hydrothermal aging at 122°C, two bar for one hour; thermal fatigue, 104 cycles between 5°C and 55°C, dwell time, 30 seconds; and mechanical fatigue, 106 cycles with a load of 70 N, sliding of 1.5 mm at 1.4 Hz; and combination of mechanical plus thermal fatigue. Fracture load was measured with a universal testing machine. Surface changes and fracture mode and origin were examined with a scanning electron microscope. Monoclinic phase content was evaluated by x-ray diffraction. The fracture load was analyzed using one-way analysis of variance at a level of 5%, and Weibull distribution was performed. No statistically significant differences were observed in the mean fracture load and characteristic fracture load among the groups (p>0.05). The Weibull modulus ranged from 6.2 to 16.6. The failure mode was similar for all groups with the crack origin located at the contact point of the indenter. Phase transformation was shown at different surfaces of the crown in all groups (1.9% to 8.9%). In conclusion, monolithic zirconia crowns possess high fracture load, structural reliability, and low phase transformation.

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Osteoporosis leads to increased bone fragility, thus effective approaches enhancing bone strength are needed. Hence, this study investigated the effect of single or combined application of high-frequency (HF) loading through whole body vibration (WBV) and alendronate (ALN) on the mechanical competence of ovariectomy-induced osteoporotic bone. Thirty-four female Wistar rats were ovariectomized (OVX) or sham-operated (shOVX) and divided into five groups: shOVX, OVX-shWBV, OVX-WBV, ALN-shWBV and ALN-WBV. (Sham)WBV loading was applied for 10 min/day (130 to 150 Hz at 0.3g) for 14 days and ALN at 2 mg/kg/dose was administered 3x/week. Finite element analysis based on micro-CT was employed to assess bone biomechanical properties, relative to bone micro-structural parameters. HF loading application to OVX resulted in an enlarged cortex, but it was not able to improve the biomechanical properties. ALN prevented trabecular bone deterioration and increased bone stiffness and bone strength of OVX bone. Finally, the combination of ALN with HF resulted in an increased cortical thickness in OVX rats when compared to single treatments. Compared to HF loading, ALN treatment is preferred for improving the compromised mechanical competence of OVX bone. In addition, the association of ALN with HF loading results in an additive effect on the cortical thickness.

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Candida albicans and streptococci of the mitis group form communities in multiple oral sites, where moisture and nutrient availability can change spatially or temporally. This study evaluated structural and virulence characteristics of Candida-streptococcal biofilms formed on moist or semidry mucosal surfaces, and tested the effects of nutrient availability and hyphal morphotype on dual-species biofilms. Three-dimensional models of the oral mucosa formed by immortalized keratinocytes on a fibroblast-embedded collagenous matrix were used. Infections were carried out using Streptococcus oralis strain 34, in combination with a C. albicans wild-type strain, or pseudohyphal-forming mutant strains. Increased moisture promoted a homogeneous surface biofilm by C. albicans. Dual biofilms had a stratified structure, with streptococci growing in close contact with the mucosa and fungi growing on the bacterial surface. Under semidry conditions, Candida formed localized foci of dense growth, which promoted focal growth of streptococci in mixed biofilms. Candida biofilm biovolume was greater under moist conditions, albeit with minimal tissue invasion, compared with semidry conditions. Supplementing the infection medium with nutrients under semidry conditions intensified growth, biofilm biovolume and tissue invasion/damage, without changing biofilm structure. Under these conditions, the pseudohyphal mutants and S. oralis formed defective superficial biofilms, with most bacteria in contact with the epithelial surface, below a pseudohyphal mass, resembling biofilms growing in a moist environment. The presence of S. oralis promoted fungal invasion and tissue damage under all conditions. We conclude that moisture, nutrient availability, hyphal morphotype and the presence of commensal bacteria influence the architecture and virulence characteristics of mucosal fungal biofilms.

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OBJECTIVE: The aim of this study was to evaluate the composition of the salivary pellicle (SP) and multispecies biofilm developed on titanium nitrided by cold plasma. METHODS: Titanium discs were allocated into a control group (Ti) and an experimental group (TiN - titanium-nitrided by cold plasma). The disc surface topography was characterized by scanning electron microscopy (SEM) and atomic force microscopy (AFM). The chemical composition of the disc surface was determined by X-ray photoelectron spectroscopy (XPS). Stimulated, clarified, and filtered saliva was used to form pellicles on the discs. Proteome analysis of the adsorbed SP proteins was performed by liquid chromatography-mass spectrometry (LC-MS). The surface free energy (SFE) was evaluated before and after SP formation. A multispecies biofilm composed of Actinomyces naeslundii, Streptococcus oralis, Streptococcus mutans, Fusobacterium nucleatum, Veillonella dispar, and Candida albicans was developed on the SP-coated discs. Viable microorganism counts were determined. The biomass and average thickness of biofilms were analyzed by confocal laser-scanning microscopy (CLSM) with COMSTAT software. The biofilm organization was visualized by SEM. RESULTS: The surface topography was similar in both groups. The SFE of the TiN group did not differ from that of the Ti group (p>0.05), although the adsorption of pellicle proteins increased the SFE in both pellicle-coated groups (p<0.001). Different proteins were identified on the Ti and TiN surfaces. The amount of biofilm was similar for both groups (p=0.416), but the counts of F. nucleatum and S. oralis were higher in the TiN group (p<0.001). Similar biofilms were characterized by the COMSTAT data, CLSM images, and SEM images. CONCLUSION: The titanium nitrided by cold plasma exhibited differences in SP composition and multispecies microbial biofilm population compared to the control titanium surface.

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The limited number of treatments for oral candidiasis resulted in the emergence of azole-resistant Candida albicans strains, thus enforcing the need for novel antifungal treatments. Although histatin 5 (H5) demonstrates antifungal activity, its inhibitory effect when adhered to hydroxyapatite and Polymetylmethacrylate (PMMA) surfaces, resembling conditions of the in vivo pellicle, remains unexplored. The objective of this in vitro study was to determine whether surface-adhered H5 inhibits the colonization of C. albicans on hydroxyapatite and/or PMMA. The C. albicans assay involved developing a mono-protein pellicle (either H5 or albumin) on hydroxyapatite and PMMA discs, introducing C. albicans and counting the number of adhered cells, throughout time, using scanning electron microscopy. A negative binomial statistical model and the Tukey-Kramer test were used for statistical analysis, with p < 0.01 indicating significance. H5-coated PMMA had significantly reduced number of cells compared to albumin-coated PMMA at 30, 90 and 1440 min (p < 0.0001), with the number of cells decreasing significantly in 90 and 1440 min (p < 0.0001). Similarly, H5-coated hydroxyapatite had significantly fewer cells compared to the albumin-coated surface at 90 and 1440 min (p < 0.0001), with the number of cells decreasing significantly at 30, 90 and 1440 min (p < 0.0001). In conclusion, C. albicans colonization was most inhibited by PMMA and hydroxyapatite-adhered H5 after 1440 min, illustrating the time-dependent effect of H5. In addition, yeast cells colonized albumin-coated PMMA, while dense hyphal networks formed on albumin-coated hydroxyapatite, suggesting that C. albicans morphology is influenced by the surface available for albumin adhesion.

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Denture liners (DL) are easily colonized by Candida spp. In an attempt to prevent biofilm colonization, manufacturers have incorporated undecylenic acid (UDA) into DL. In this in vitro study, the effects of UDA released from DL on Candida biofilms were investigated. The concentrations of UDA released from commercial DL were determined by gas chromatography-mass spectrometry (GC-MS). Minimum inhibitory concentration (MIC) and minimum fungistatic concentration (MFC) tests were performed for C. albicans or C. glabrata, with UDA for comparison with the concentrations released from DL. Specimens of DL with (experimental group) and without UDA (control group) were fabricated, and Candida biofilms were developed on DL surfaces. Biofilms were evaluated by cell counts, metabolic activity, structure, and secretion of proteinase or phospholipase. The concentrations of UDA released were within the MIC and MFC ranges. In the presence of UDA, C. albicans biofilms were thinner and had lower numbers of viable and active cells, although no significant enzymatic changes were observed relative to the control group (p > 0.05). In contrast, C. glabrata biofilms exhibited higher cell counts and greater metabolic activity and also increased proteinase activity in the presence of UDA relative to the control group (p < 0.05). Overall, UDA did not prevent Candida biofilm formation.

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To test the effect of distinct solubilities of dentine and enamel on mineral ion concentration in the biofilm fluid during a sugar-induced pH drop, dental biofilms were formed in situ for 4 days on acrylic (control), dentine or enamel. On the 5th day, they were treated with water (control) or 20% glucose and collected 5 min later. Significantly lower pH values and higher calcium concentrations were found in the biofilm fluid after glucose exposure, without significant differences among the three substrates. During pH drop, biofilm reservoirs release calcium to the fluid, masking the differential solubility between enamel and dentine.

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Non-carious cervical lesions (NCCLs) are a group of lesions that affect the cervical area of the teeth. Different aetiological factors have been reported, among them tooth brushing force, erosive agents and occlusal forces. The aim of this study was to ascertain, by means of a systematic review, the association between NCCLs and occlusion. A search was performed in the MEDLINE database, retrieving a total of 286 articles. After title and abstract screening, the clinical investigations were read in full to select those that evaluated the occlusal aspects of NCCL aetiology. This systematic review describes the results of three prospective and 25 cross-sectional studies. As there is an extreme heterogeneity in design, diagnostic criteria, forms of analysis and associated factors, a meta-analysis was not possible. The wide variety of classification and diagnostic criteria reflects a high range of reported prevalence. An extensive heterogeneity of independent variables was noted, even in occlusal analysis, which helps to mask any conclusion about the role of occlusion in NCCL aetiology. The literature reveals that studies on this topic are subject to a substantial amount of bias, such as evaluation (use of non-blinded examiners) and confounding bias (no control of others aetiological factors). Up to now, it has been impossible to associate NCCLs with any specific causal agent, and the role of occlusion in the pathogenesis of non-carious cervical lesions seems as yet undetermined. Therefore, additional studies, properly designed to diminish bias, are warranted.

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