The Synergistic Effect of High Intensity Focused Ultrasound on In-vitro Remineralization of Tooth Enamel by Calcium Phosphate Ion Clusters.

Background: Remineralization of dental enamel is an important intervention strategy for the treatment of demineralized lesions. Existing approaches have limitations such as failure to adequately reproduce both the ideal structural and mechanical properties of the native tooth. The ability of ultrasound to control and accelerate the crystallization processes has been widely reported. Therefore, a new approach was explored for in-vitro enamel remineralization involving the synergistic effect of high-intensity focused ultrasound (HIFU) coupled with calcium phosphate ion clusters (CPICs). Methods: The demineralized enamel was treated with CPICs, with or without subsequent HIFU exposure for different periods (2.5, 5, and 10 min). The specimens were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), and Raman spectroscopy. The surface hardness and crystallographic properties of the treated specimens were evaluated using Vickers microhardness testing and X-ray diffraction (XRD), respectively. Results: SEM revealed distinct, organized, and well-defined prismatic structures, showing clear evidence of remineralization in the combined CPIC/HIFU treatment groups. AFM further revealed a decrease in the surface roughness values with increasing HIFU exposure time up to 5 min, reflecting the obliteration of interprismatic spaces created during demineralization. The characteristic Raman band at 960 cm-1 associated with the inorganic phase of enamel dominated well in the HIFU-treated specimens. Importantly, microhardness testing further demonstrated that new mineral growth also recovered the mechanical properties of the enamel in the HIFU-exposed groups. Critical to our aspirations for developing this into a clinical process, these results were achieved in only 5 min. Conclusion: HIFU exposure can synergise and significantly accelerate in-vitro enamel remineralization process via calcium phosphate ion clusters. Therefore, this synergistic approach has the potential for use in future clinical interventions.

Evaluation of Antibacterial Efficacy of High-Intensity Focused Ultrasound Versus Photodynamic Therapy Against Enterococcus faecalis-Infected Root Canals.

OBJECTIVE: The high incidence of endodontic failure is associated with the remnants of Enterococcus faecalis present within the intricate anatomies of the root canal system (RCS), often inaccessible by the current endodontic practices. This study was aimed at evaluating the effect of high-intensity focused ultrasound (HIFU) and photodynamic therapy (PDT) on E. faecalis biofilms in artificially infected root canals for the potential application in current endodontic practices. METHODS: Forty-five single-rooted extracted teeth were instrumented using hand files, sterilized in an autoclave, infected with E. faecalis and incubated for 4 wk. The specimens were treated and identified as follows: Control, 4% sodium hypochlorite (NaOCl); riboflavin (1 mg/mL); light only; HIFU (250 kHz, 20 W, 60s); PDT; riboflavin/HIFU; light/HIFU; and riboflavin/HIFU/light. Bactericidal efficacy was determined by colony-forming units (CFU), (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) (MTT) assay, scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). RESULTS: Enterococcus faecalis biofilm exhibited significantly lower metabolic activity when treated with HIFU (250 kHz, 20 W, 60 s) compared with the control (4% NaOCl) and PDT groups. A similar phenomenon was observed with the CFU assay. HIFU remained the most effective treatment modality, with consistent results in CLSM and SEM. CONCLUSION: This study highlighted the potential application of HIFU as an adjunct drug-free, non-destructive root canal disinfection method for endodontic treatment, suggesting an alternative to the current gold standard of 4% NaOCl and PDT.

Development of 3D printed dental resin nanocomposite with graphene nanoplatelets enhanced mechanical properties and induced drug-free antimicrobial activity.

OBJECTIVES: Oral prosthetic rehabilitation has been used for a long time to restore function and natural appearance; however, it is still one of the most challenging areas in dentistry due to its technical fabrication process and biological behavior. Considering the advantages of additive manufacturing technology, this study introduced the feasibility of developing a 3D printed resin-based composition modified with graphene nanoplatelets (GNPs) to improve properties. METHODS: Acrylate-based resin was impregnated with different concentrations of GNPs (0.0-0.25 wt%), and then different aspects such as mechanical, physical, biological and antimicrobial were analyzed to evaluate the effectiveness. TEM and SEM were used to characterize GNPs and their existence within the resin. Surface topography and roughness were evaluated using AFM. The degree of conversion and composition were confirmed by FTIR. Mechanical properties were detected using bending strength, microhardness and nanoindentation. Biocompatibility and antimicrobial activities were assessed with oral fibroblast and Candida albicans (C. albicans), respectively. In addition, most of the measurements were performed repeatedly after 3 months of storage in artificial saliva to evaluate performance. RESULTS: GNPs improved strength significantly at low concentrations ≤ 0.05 wt%, while the addition up to 0.25 wt% enhanced printed nanocomposite hardness and elasticity. The modification did not induce a toxic response, as its biocompatibility was within the recommended range of biomedical devices. Antimicrobial activity was of prominence, as GNPs showed an outstanding route of reducing C. albicans activity associated with filler proportion. SIGNIFICANCE: The embedment of GNPs in 3D printed resin can become a key material for customized applications that require high antimicrobial, stiffness and strength properties.

Silver-loaded mesoporous silica nanoparticles enhanced the mechanical and antimicrobial properties of 3D printed denture base resin.

The aim of this study is to develop a novel 3D printed denture base resin material modified with mesoporous silica nanocarrier loaded with silver (Ag/MSN) to enhance mechanical and antimicrobial properties. Acrylate resin-based was incorporated with various proportion of Ag/MSN (0.0-2.0 wt%). Specimens with different geometry were printed and characterized accordingly for the effect of modification on properties such as: mechanical and physical properties, chemical composition and degree of conversion, as well as biological response in term of biocompatibility and antimicrobial against oral fibroblast and candida biofilm (C. albicans), respectively. The consecutive addition of Ag/MSN improved significantly surface hardness and crack propagation resistance, while flexural strength remained similar to control; however, a negligible decrease was observed with higher concentrations ≥1 wt%. No significant difference was noticed with water sorption, while water solubility had a remarkable trend of reduction associated with filler content. The surface roughness significantly increased when concentration of Ag/MSN was ≥1.0 wt%. A significant reduction in C. albicans biofilm mass, as the inhibition proficiency was correlated with the proportion of the filler. With respect to the amount of Ag/MSN, the modification was compatible toward fibroblast cells. The sequential addition of Ag/MSN enhanced significantly the mechanical and antimicrobial properties of the 3D printed resin-based material without affecting adversely compatibility. The acrylic resin denture base material has susceptibility of microbial adhesion which limits its application. Silver loaded MSN showed a significant performance to enhance antimicrobial activity against C. albicans which is the main cause of denture stomatitis. The proposed invention is a promise technique for clinical application to provide an advanced prosthesis fabrication and serve as long-term drug delivery.

Cytotoxicity and antimicrobial efficiency of ZrO2 nanoparticles reinforced 3D printed resins.

OBJECTIVE: The aim of this study was to investigate the potential antimicrobial and cytotoxic effect of modified 3D printed resin with ZrO2 nanoparticles, as long-term provisional restoration. In addition, the study involved artificial aging process for three months to observe stability of 3D printed resin. METHODS: Functionalized ZrO2 nanoparticles with γ-MPS were characterized using transmission electron microscopy, scanning electron microscope and Fourier-transform infrared spectroscopy. Dental resin was incrementally impregnated with γ-MPS modified nanoparticles at different concentrations (0, 1, 3, and 5 wt%). Specimens were printed, post-cured and placed in artificial saliva at 37 oC for 48 h or aged for 3 months. Discrepancy in composition and roughness were monitored using FTIR and AFM, respectively. Biocompatibility was evaluated using human oral fibroblasts. Antimicrobials capacity and biofilm adhesion were measured with Streptococcus mutans and Candida albicans. RESULTS: The microscopic and spectroscopic analyses confirmed γ-MPS coating around ZrO2 nanoparticles. The addition of nanoparticles (>1 wt%) significantly increased the surface roughness. Cytotoxicity results were in agreement with the recommended range of oral biomaterials standard. Moreover, the antimicrobial activity significantly improved with increasing the filler concentration. Despite the decrease in antimicrobial efficacy after 3 months of aging, modified resin revealed a critical ability to dominate biofilm formation. SIGNIFICANCE: The addition of ZrO2 nanoparticles showed significant antimicrobial capability of a 3D printed resin without inducing any cellular side effects. Thus, the modification of a 3D printed resin with ZrO2 nanoparticles has a promising future in the dental field for fabricating long-term provisional restorations.

Effect of post-curing light exposure time on the physico-mechanical properties and cytotoxicity of 3D-printed denture base material.

OBJECTIVE: This study investigated the effect of post curing light exposure time on the physico-mechanical properties and cytotoxicity of a 3D-printed PMMA-based denture material in comparison to a conventional heat-cured alternative as a control. METHODS: 3D-printed specimens were fabricated followed by post-curing for 0, 5, 10 or 20 min at 200 W and light wavelength range of 390-540 nm. Heat-cured specimens were fabricated using a standard protocol. Specimens were placed in artificial saliva at 37 â„ƒ for 48 h (immediate groups) and 6 months (aged group), then evaluated flexural strength/modulus, fracture toughness, microhardness, and degree of conversion. Water sorption and solubility was assessed after 28 days. Flexural strength, flexural modulus, and fracture toughness were tested through three-point bending tests, while the surface hardness was tested using Vickers's test. Fractured specimens were viewed by scanning electron microscope (SEM). Cytotoxicity in term of cell viability was evaluated using human oral fibroblasts. RESULTS: Flexural strength/modulus, fracture toughness and surface hardness significantly improved with the increase in light curing time up to 20 min. The same pattern of improvement was found with degree of conversion, water sorption, solubility, and cell viability. There was no significant difference (p < 0.01) between heat-cured material and 3D specimens post-cured for 20 min in term of flexural strength/modulus, surface hardness, and degree of conversion at the two-storage time points. SIGNIFICANCE: Generally, the physico-mechanical properties of the 3D-printed denture base material improve as post curing time increases up to 20 min which exhibited comparable performance as the conventional heat-cured control.

Silanization of nanographene platelets improves interaction with the dentin bonding resin matrix and enhances interfacial bond integrity to dentin.

This study synthesized and characterized graphene nanoplatelets silanized with 3-(trimethoxysilyl)propyl methacrylate (MPS-GNP) for morphological and chemical characteristics. In addition, we modified a dentin bonding agent using different concentrations of MPS-GNP to study its interaction within the resin matrix of the adhesive, degree of conversion (DC), biological, and mechanical properties after bonding to tooth. Both 0.25% and 0.5% MPS-GNP-modified bonding agents showed comparable DC values to the unmodified control adhesive (range: 41%-43%). However, a statistically significant reduction in the DC was found when 0.25% and 0.5% non-silanized GNP was doped with the adhesive (<38%) (p < 0.05). On day 30, the bacterial viability of 0.5% GNP and MPS-GNP groups remained very low under 22% with the highest dead cell count (p < 0.05). GNP incorporated within the resin matrix of the dentin bonding agent showed clear evidence of several interfacial gap formations and non-union between the GNP surface and resin matrix, while the MPS-GNP modified dentin bonding agent showed MPS-GNP with no gap formation with complete union between the graphene surface and resin matrix. The decrease in the µTBS was least pronounced for 0.25% and 0.5% MPS-GNP groups. After 12 months of ageing, the groups 0.25% and 0.5% MPS-GNP also showed the highest BS as compared to the rest of the groups. Statistically significant reduction was seen in nanohardness at the hybrid layer and adhesive layer for GNP groups after 4 months of storage. The addition of up to 0.5% MPS-GNP showed optimized DC, antibiofilm activity, and micro-tensile bond strength without affecting the standard adhesion characteristics as compared to GNP alone.

Adapting World Health Organization Disability Assessment Schedule 2.0 for Nepal.

BACKGROUND: Disability is a vital public health issue for health care programs. Affluent countries usually prioritize disability-related research, while often it remains neglected in resource-poor countries like Nepal. The aim of this study was to make available a translated and culturally adapted version of the World Health Organization Disability Assessment Schedule 2.0 (WHODAS 2.0) for measuring disability in the Nepalese population. METHODS: WHODAS 2.0 (12-items version) was translated into Nepali using a standard forward-backward translation protocol. Purposive and convenience recruitment of participants with psychiatric disabilities was done at the Psychiatry services in a tertiary care hospital. Age and gender-matched participants with physical disabilities were selected from the Internal Medicine department, and participants with no disability were recruited from their accompanying persons. A structured interview in Nepali including the translated WHODAS 2.0 was administered to all participants. Exploratory factor analysis and parallel analysis assessed the construct validity. Content validity was explored, and a quality of life instrument was used for establishing criterion validity. Reliability was measured via Cronbach alpha. Mann-Whitney test explored score differences between the disabled and non-disabled. RESULTS: In total, 149 persons [mean age: 40.6 (12.8); 43.6% males, 56.4% females; 61.7% disabled, 38.3% non-disabled] consented to participate. Parallel analysis indicated that a single factor was adequate for the Nepali WHODAS version that captured 45.4% of the total variance. The translated scale got a good Cronbach alpha (= 0.89). Satisfactory construct, content and criterion validity was found. The WHODAS total scores showed a significant difference between the disabled and non-disabled (U = 2002.5; p = 0.015). However, the difference between psychiatric and physical disabilities was not significant, which underscores that the scale is rating disability in general. CONCLUSION: The one-factor structure of the translated and culturally adapted Nepali-version of WHODAS 2.0 showed acceptable validity and an adequate reliability. For epidemiological research purposes, this version of WHODAS 2.0 is now available for measuring global disability in Nepal.

Depression and Quality of Life among the Chronic Kidney Disease Patients.

BACKGROUND: Depression and other mental illness are very common among chronic kidney disease and also Quality of life has been found significantly impaired in chronic kidney disease. The objective of our study is to study Depression, Quality of life and its associations in chronic kidney disease. METHODS: This was a descriptive cross-sectional study. We used convenient method of sampling for data collection. The World Health Organization Quality-of-Life 8-question scale was used for estimating quality of life and Beck Depression Inventory  was used for the assessment of depression.  Independent samples t-test was conducted to analysed bivariate relationship of sociodemographic factors with depression and Quality of life scores and multiple linear regression analysis was performed to determine predictors of Quality of life. RESULTS: The 75.5% participants found to have depression. Depression was found statistically significant across socioeconomic status (p value 0.04) and other medical comorbidities (p value 0.04). Variables found to be significantly associated with total quality of life in multiple linear regression analysis were caste (p value 0.03), socioeconomic status (p value 0.009) and depression (p value 0.001). CONCLUSIONS: Depression and reduced quality of life is very common among chronic kidney patients. Low socioeconomic status and comorbid medical conditions were associated with depression and caste, low socioeconomic status and depression were associated with low quality of life. Screening and management of depression should be included in the routine care and it could help improving the quality of life of patients with chronic kidney disease.

Periodontal Health Status and Pregnancy Outcomes: A Survey in Medical Doctors.

INTRODUCTION: Periodontal disease and caries are the most common causes of tooth loss worldwide. Studies have demonstrated strong association between periodontitis and adverse pregnancy outcomes. Medical doctors, who are the primary healthcare providers, seldom advise women to seek dental care during pregnancy. This study was undertaken to explore the knowledge, attitudes and behaviours of medical doctors towards oral health and to identify the barriers of prenatal periodontal healthcare in their practices and its possible implications on pregnancy outcomes. METHODS: Total 377 doctors filled the questionnaire. The data collected through personal contacts, social networking, emails, online forms and networking at conferences were analysed using Statistical Package for the Social Sciences 20 software program and presented in tables, charts and diagrams. RESULTS: Out of 263 (69.8%) male and 114 (30.2%) female doctors enrolled in the study, only 52 (13.8%) had received education or training on oral care during pregnancy. Among them 299 (79.3%) agreed that there is possible link between health of teeth-gums and pregnancy. Approximately 105 (27.9%) encountered patients with oral/periodontal problem every week but only 108 (28.6%) "always" advised their patient for regular dental check-ups. Similarly, 358 (95%) agreed that there is need for universal guidelines however, 133 (35.3%) thought there was insufficient time to advise patients on oral health during check-ups. CONCLUSIONS: There is need for training on 'oral healthcare during pregnancy' for medical doctors. Developing universal guidelines for oral healthcare in pregnant women for all health professionals would be another important step. An adequate referral system to oral healthcare providers and biannual check-ups is recommended for both general patient as well as pregnant women for preventing adverse situations related to oral and specifically periodontal diseases.

Widespread intra-dependencies in the removal of introns from human transcripts.

Research into the problem of splice site selection has followed a reductionist approach focused on how individual splice sites are recognized. Early applications of information theory uncovered an inconsistency. Human splice signals do not contain enough information to explain the observed fidelity of splicing. Here, we conclude that introns do not necessarily contain 'missing' information but rather may require definition from neighboring processing events. For example, there are known cases where an intronic mutation disrupts the splicing of not only the local intron but also adjacent introns. We present a genome-wide measurement of the order of splicing within human transcripts. The observed order of splicing cannot be explained by a simple kinetic model. Simulations reveal a bias toward a particular, transcript-specific order of intron removal in human genes. We validate an extreme class of intron that can only splice in a multi-intron context. Special categories of splicing such as exon circularization, first and last intron processing, alternative 5 and 3'ss usage and exon skipping are marked by distinct patterns of ordered intron removal. Excessive intronic length and silencer density tend to delay splicing. Shorter introns that contain enhancers splice early.

Large-scale analysis of branchpoint usage across species and cell lines.

The coding sequence of each human pre-mRNA is interrupted, on average, by 11 introns that must be spliced out for proper gene expression. Each intron contains three obligate signals: a 5' splice site, a branch site, and a 3' splice site. Splice site usage has been mapped exhaustively across different species, cell types, and cellular states. In contrast, only a small fraction of branch sites have been identified even once. The few reported annotations of branch site are imprecise as reverse transcriptase skips several nucleotides while traversing a 2-5 linkage. Here, we report large-scale mapping of the branchpoints from deep sequencing data in three different species and in the SF3B1 K700E oncogenic mutant background. We have developed a novel method whereby raw lariat reads are refined by U2snRNP/pre-mRNA base-pairing models to return the largest current data set of branchpoint sequences with quality metrics. This analysis discovers novel modes of U2snRNA:pre-mRNA base-pairing conserved in yeast and provides insight into the biogenesis of intron circles. Finally, matching branch site usage with isoform selection across the extensive panel of ENCODE RNA-seq data sets offers insight into the mechanisms by which branchpoint usage drives alternative splicing.

Involvement of co-repressor LUH and the adapter proteins SLK1 and SLK2 in the regulation of abiotic stress response genes in Arabidopsis.

BACKGROUND: During abiotic stress many genes that are important for growth and adaptation to stress are expressed at elevated levels. However, the mechanisms that keep the stress responsive genes from expressing under non stress conditions remain elusive. Recent genetic characterization of the co-repressor LEUNIG_HOMOLOG (LUH) and transcriptional adaptor proteins SEUSS-LIKE1 (SLK1) and SLK2 have been proposed to function redundantly in diverse developmental processes; however their function in the abiotic stress response is unknown. Moreover, the molecular functions of LUH, SLK1 and SLK2 remain obscure. Here, we show the molecular function of LUH, SLK1 and SLK2 and the role of this complex in the abiotic stress response. RESULTS: The luh, slk1 and slk2 mutant plants shows enhanced tolerance to salt and osmotic stress conditions. SLK1 and SLK2 interact physically with the LUFS domain in LUH forming SLK1-LUH and SLK2-LUH co-repressor complexes to inhibit the transcription. LUH has repressor activity, whereas SLK1 and SLK2 function as adaptors to recruit LUH, which in turn recruits histone deacetylase to the target sequences to repress transcription. The stress response genes RD20, MYB2 and NAC019 are expressed at elevated levels in the luh, slk1 and slk2 mutant plants. Furthermore, these stress response genes are associated with decreased nucleosome density and increased acetylation levels at H3K9 and H3K14 in the luh, slk1 and slk2 mutant plants. CONCLUSIONS: Our results indicate that SLK1, SLK2 and LUH form a co-repressor complex. LUH represses by means of an epigenetic process involving histone modification to facilitate the condensation of chromatin thus preventing transcription at the target genes.