Investigation of the crystal formation from calcium silicate in human dentinal tubules and the effect of phosphate buffer saline concentration.

Background/purpose: Based on hydrodynamic theory, blocking the dentinal tubules can reduce discomfort caused by dentin hypersensitivity. This study identified the crystals formed in dentinal tubules from tricalcium silicate (TCS) in phosphate-buffered saline (PBS) and evaluated the effect of PBS concentration on crystal formation. Materials and methods: Sixty-nine specimens were made by isolating the cervical part of extracted premolars. TCS was applied by brushing for 10,000 strokes on dentin surface simulating sensitive dentin. Specimens were stored in PBS or solutions with concentrations 1/100, 1/10, 10, and 100 times that of PBS for 1, 30, 60, or 90 days (n = 3). Another nine specimens applied TCS, were immersed in PBS for 3 months, and divided into three subgroups: no treatment, sonication for 10 min, and 1M acetic acid treatment for 3 min. Crystal formation was examined using a scanning electron microscope, assigned five grade scores (0-4) according to maturation, and analyzed by a nonparametric two-way ANOVA (α = 0.05). Crystal components were analyzed using X-ray diffraction (XRD) and energy dispersion X-ray spectroscopy (EDS). Results: The maturation of intratubular crystals was dependent on time and PBS concentration (P < 0.05). In all periods, the high-concentration group showed a higher maturation grade than the low-concentration group. Intratubular crystals were similar to hydroxyapatite according to XRD and EDS, and they withstood sonication and acid application. Conclusion: TCS with nanosized particles formed hydroxyapatite-like crystals in the dentinal tubules, which were dependent on time and concentration of PBS and withstood sonication and acid application.

Transcriptomic analysis of sheep hypothalamus discloses regulatory genes potentially involved in sex-dependent differences in body weight of progeny born to dams supplemented with omega-3 fatty acids or methionine during late-gestation.

Fetal programming research conducted in sheep has reported sexually dimorphic responses on growth of the progeny born to in-utero methionine or omega-3 fatty acids supplementation. However, the biological mechanism behind the nutrient by sex interaction as a source of variation in offspring body weight is still unknown. A high-throughput RNA sequencing data of hypothalamus samples from 17 lambs were used in the current study to identify differentially expressed genes (DEGs) between males and females born to dams supplemented with different nutrients during late-gestation. Ewes received a basal diet without omega-3 fatty acids or methionine supplementation as the control (CONT); omega-3 fatty acids supplementation (FAS), or methionine supplementation (METS). A list of regulated genes was generated. Data were compared as CONT vs. FAS and CONT vs. METS. For CONT vs. METS, a treatment by sex interaction was found (adjusted P-value < 0.05) on 121 DEGs (112 upregulated and 9 downregulated) on female lambs born to METS compared with METS males. Importantly, with the sex interaction term, more than 100 genes were upregulated in female lamb's hypothalamuses born to METS. Gene Ontology (GO) and Ingenuity Pathway Analysis (IPA) were performed using the DEGs from female lambs. Terms under biological process (related to morphogenesis, organism, and tissue development), cellular component (related to chromatin, extracellular components), and molecular function (involved in chromatin structure and transcription and factors linked to binding DNA) were presented (adjusted P-value < 0.05) for GO. For the IPA, the top-scoring network was developmental disorder, endocrine system development and function, and organ morphology. Only a few differences were observed in the comparison between the interaction of sex and treatment for the CONT vs. FAS comparison. The markedly increased number of DEGs substantially involved in developmental and growth processes indicates the extent to which maternal methionine supplementation causes the sexually dimorphic effects observed in the offspring.

Feeding dams during gestation affects the development of the offspring for their entire life. The objective of the current experiment was to evaluate the changes of the transcriptome in the hypothalamus of the offspring lambs born from dams supplemented with (i) a control diet (without lipids or methionine supplementation), (ii) an omega-3 fatty acid supplementation, or (iii) a methionine supplementation. The supplementation took place in the last third of gestation and the hypothalamus of male and female offspring was collected after being on a fattening diet for 54 d. Hypothalamus samples were used to extract RNA and analyzed using RNA sequencing. There was an interaction due to sex and methionine supplementation. The pathways that were modified were chromatin structure, developmental processes, and organ morphology. The modification observed on these pathways could explain the sex by treatment interaction differences previously observed in growth. There were few sex by omega-3 fatty acid interactions on the hypothalamus transcriptome. Therefore, the sexual dimorphism observed by methionine supplementation may be regulated by the hypothalamus.

Correlation of two different devices for the evaluation of primary implant stability depending on dental implant length and bone density: An in vitro study.

Non-invasive objective implant stability measurements are needed to determine the appropriate timing of prosthetic fitting after implant placement. We compared the early implant stability results obtained using resonance frequency analysis (RFA) and damping capacity analysis (DCA) depending on the implant length and bone density. Total 60, 4.0 mm diameter implants of various lengths (7.3 mm, 10 mm, and 13 mm) were used. In Group I, low-density bone was described using 15 PCF (0.24 g/cm3) polyurethane bone blocks, and in Group II, 30 PCF (0.48 g/cm3) polyurethane bone blocks were used to describe medium density bone. RFA was performed using an Osstell® Beacon+; DCA was performed using Anycheck®. Measurements were repeated five times for each implant. Statistical significance was set at P <0.05. In Group I, bone density and primary implant stability were positively correlated, while implant length and primary implant stability were positively correlated. In Group II, the implant stability quotient (ISQ) and implant stability test (IST) values in did not change significantly above a certain length. Primary implant stability was positively correlated with bone density and improved with increasing implant length at low bone densities. Compared with the Osstell® Beacon+, the simplicity of Anycheck® was easy to use and accessible.

The Association of Job Stress, Quality of Sleep, and the Experience of Near-Miss Errors among Nurses in General Hospitals.

This study investigated the association of job stress, quality of sleep, and near-miss error experiences among nurses working in general hospitals. A convenience sample of 195 nurses with over 3 months of work experience in a general hospital participated in the study. Statistical analyses were performed using SPSS 27.0. Factors influencing experiences of near-miss errors were examined through univariable and multivariate logistic regression. In total, 58.5% of nurses in a general hospital had experienced near-miss errors. Nurses in the upper job stress tertile (≥118) were 2.24 times more likely to encounter near-miss errors (95% CI: 1.07-4.69) and particularly 2.58 times (95% CI: 1.26-5.26) in falls and medical equipment-related near-miss errors. Nurses working more than 3 h of overtime per week had a 2.72 times higher (95% CI: 1.35-5.48) likelihood of experiencing near-miss errors related to falls and medical equipment and 4.03 times higher (95% CI: 1.92-8.45) likelihood of experiencing near-miss errors associated with examination procedures. To prevent near-miss errors among nurses in general hospitals, effective management of organizational job stress is crucial. Particularly for departments with a high level of overtime work (more than 3 h/week), it is essential to provide and manage educational programs on patient safety.

Comparative analysis of flexural strength of 3D printed and milled 4Y-TZP and 3Y-TZP zirconia.

STATEMENT OF PROBLEM: The mechanical properties of 3 dimensionally (3D) printed zirconia have been reported to be comparable with those of milled zirconia, except for the flexural strength. However, most previous studies tested 3 mol% yttria-stabilized tetragonal zirconia polycrystal (3Y-TZP), making it necessary to study 3D printed zirconia with 4 mol% yttria content (4Y-TZP). PURPOSE: The purpose of this in vitro study was to compare the flexural strength of 3D printed 4Y-TZP with 3Y-TZP materials and milled 4Y-TZP. MATERIAL AND METHODS: A total of 80 disk specimens (Ø15×1.5 mm) were fabricated and divided into 4 groups (n=20) using the fabrication method and yttria content: milled 3Y-TZP (Katana HT; Kuraray Noritake), 3D printed 3Y-TZP (TZ-3Y-E; Tosoh), milled 4Y-TZP (Katana STML; Kuraray Noritake), and 3D printed 4Y-TZP (3DMAT; Genoss). The biaxial flexural strength was determined with a piston-on-3-ball test (n=15). The flexural strength of each specimen was measured, and the Weibull modulus (m) and characteristic strength (σ0) were estimated from the fracture load distribution. Two intact and fractured specimens were examined with scanning electron microscopy (SEM). The crystalline phase of the specimens in each group was identified through X-ray diffraction (XRD) analysis (n=5). A 1-way ANOVA was used to compare the flexural strength among different groups. Subsequently, pairwise comparisons were conducted with the Tukey post hoc method (α=.05). RESULTS: The flexural strength of 3D printed 4Y-TZP was significantly higher than that of milled 4Y-TZP (P<.001). In contrast, the flexural strength of 3D printed 3Y-TZP was significantly lower than that of milled 3Y-TZP (P<.001). X-ray diffraction (XRD) analysis revealed that the tetragonal phase was the dominant phase in all groups, with the identification of some cubic phase peaks. CONCLUSIONS: Three dimensionally printed 4Y-TZP showed significantly higher flexural strength than milled 4Y-TZP and exhibited a clinically acceptable flexural strength exceeding 800 MPa.

Association between dental diseases and oral hygiene care and the risk of vertebral fracture: a nationwide cohort study.

Periodontal disease and increased missing teeth were associated with incident vertebral fractures. In contrast, professional dental cleaning and frequent tooth brushing, was associated with a lower risk of vertebral fracture. Better oral hygiene care attenuated the risk associated with dental diseases. PURPOSE: To investigate the association between oral health and the risk of vertebral fractures. METHODS: We included 2,532,253 individuals aged ≥40 years who underwent the Korean National Health Insurance Service health examinations in 2008 and followed up until December 31, 2017. We performed multivariable Cox proportional hazard regression analyses to evaluate the association between dental diseases and oral hygiene care and the risk of vertebral fractures. RESULTS: Over the 9.3-year median follow-up, 1.46% (n = 36,857) experienced vertebral fractures. Individuals with dental diseases had a higher risk of vertebral fracture than those without (hazard ratio [HR] 1.04, 95% confidence interval [CI]: 1.02-1.07 for periodontal diseases; 1.02, 1.00-1.05 for dental caries; 1.12, 1.05-1.20 for ≥15 missing teeth). Good oral hygiene care was associated with a lower vertebral fracture risk (HR 0.89, 95% CI: 0.86-0.91 for ≥1 time/year [vs. <1 time/year] of professional dental cleaning; 0.90, 0.87-0.93 for ≥2 times/day [vs. 0-1 time/day] of toothbrushing). The combined dental diseases was significantly associated with an increased vertebral fracture risk, whereas combined oral hygiene care was associated with further risk reduction. Better oral hygiene care reduced vertebral fracture risk associated with dental diseases (all P <0.001). CONCLUSION: Periodontal disease, dental caries, and an increased number of missing teeth were independently associated with higher risks for vertebral fractures. Conversely, improved oral hygiene care, such as personal dental cleaning and frequent tooth brushing, may modify vertebral fracture risks associated with dental disease.

The AIRN lncRNA is imprinted and paternally expressed in pigs.

Genomic imprinting plays critical roles during the development of mammalian species and underlying epigenetic mechanisms frequently involve long non-coding RNAs (lncRNAs). The paternal transcription of the antisense Igf2r RNA noncoding (Airn) is responsible for paternal silencing of the mouse insulin-like growth factor 2 receptor (Igf2r) gene and maternal Igf2r expression. Although the corresponding maternal DNA methylation imprint is conserved in humans and pigs, the orthologous AIRN lncRNA has been identified in humans but not in pigs. Here, we aimed to examine imprinted allelic expression of the porcine AIRN lncRNA along with a corresponding differentially methylated region (DMR) and to analyze allelic expression of AIRN and IGF2R in pigs. By comparing parthenogenetic and control porcine embryos, we identified a maternally methylated DMR and a significantly higher expression of AIRN lncRNA in control embryos (P < 0.05) indicating its paternal expression. Further analyses revealed that the expression of AIRN lncRNA was enriched in the pig brain and its subregions, and it was monoallelically expressed; whereas, IGF2R was expressed biallelically suggesting an absence of allele-specific transcriptional regulation. Our findings will lead to further investigations into the role of the imprinted porcine AIRN lncRNA during pig development.

Genomic imprinting is important for the development of mammals and long non-coding RNAs are often involved in the imprinting process. In mice, Airn encodes a long non-coding RNA that is imprinted, and therefore, transcribed only from the paternal allele. This paternal transcription of Airn interferes with the adjacent Igf2r promoter, leading to maternal expression of Igf2r. In pigs, the orthologous AIRN has not been identified as well as its imprinting. In the current study, we report porcine AIRN and allelic expression of both AIRN and IGF2R using our parthenogenetic embryo models and various normal pig tissues.

Imprinting at the KBTBD6 locus involves species-specific maternal methylation and monoallelic expression in livestock animals.

BACKGROUND: The primary differentially methylated regions (DMRs) which are maternally hypermethylated serve as imprinting control regions (ICRs) that drive monoallelic gene expression, and these ICRs have been investigated due to their implications in mammalian development. Although a subset of genes has been identified as imprinted, in-depth comparative approach needs to be developed for identification of species-specific imprinted genes. Here, we examined DNA methylation status and allelic expression at the KBTBD6 locus across species and tissues and explored potential mechanisms of imprinting. RESULTS: Using whole-genome bisulfite sequencing and RNA-sequencing on parthenogenetic and normal porcine embryos, we identified a maternally hypermethylated DMR between the embryos at the KBTBD6 promoter CpG island and paternal monoallelic expression of KBTBD6. Also, in analyzed domesticated mammals but not in humans, non-human primates and mice, the KBTBD6 promoter CpG islands were methylated in oocytes and/or allelically methylated in tissues, and monoallelic KBTBD6 expression was observed, indicating livestock-specific imprinting. Further analysis revealed that these CpG islands were embedded within transcripts in porcine and bovine oocytes which coexisted with an active transcription mark and DNA methylation, implying the presence of transcription-dependent imprinting. CONCLUSIONS: In this study, our comparative approach revealed an imprinted expression of the KBTBD6 gene in domesticated mammals, but not in humans, non-human primates, and mice which implicates species-specific evolution of genomic imprinting.

Effects of ultraviolet weathering aging on the color stability and biocompatibility of various computer-aided design and computer-aided manufacturing glass-ceramic materials.

OBJECTIVES: This study assessed the changes in color stability and biocompatibility of computer-aided design and computer-aided manufacturing (CAD-CAM) glass-ceramics after ultraviolet weathering (UW) aging. METHODS: A total of 300 plate-shaped specimens (12.0 × 14.0 × 1.5 mm3) were prepared using a leucite-reinforced glass-ceramic (IPS Empress CAD; E), a lithium disilicate (IPS e.max CAD; M), and two zirconia-reinforced lithium silicate (Celtra Duo; C, Vita Suprinity; V) glass-ceramics. Specimens were divided into three groups (n = 25, each), subjected to water storage at 37 °C for 24 h (control group), or UW aging at 150 kJ/m2 (first-aged group) or 300 kJ/m2 (second-aged group). The color stability, mechanical and surface properties, and biocompatibility of the CAD-CAM glass-ceramics were investigated experimentally, followed by statistical analysis. RESULTS: The brightness and redness or greenness were reduced in all groups after aging. After the first aging, V exhibited the largest color change and E exhibited the smallest color change. After the second aging, E exhibited the highest nanoindentation hardness and Young's modulus. The surface roughness was the highest for V after the first aging. Furthermore, the hydrophilicity of the materials increased after aging process. The cell proliferation/viability of human gingival fibroblasts was the highest in E before and after aging. Almost all cells survived for all groups based on a live/dead assay. CONCLUSIONS: Leucite-reinforced glass-ceramic exhibit the highest color stability and biocompatibility after aging. The color stability and biocompatibility of CAD-CAM glass-ceramics depend on the aging process and material type. CLINICAL SIGNIFICANCE: Various CAD-CAM glass-ceramics exhibit adequate color stability after UW aging. The leucite-reinforced glass-ceramics exhibit the highest color stability, cell proliferation, and viability after aging. The color stability, mechanical and surface properties, and biocompatibility of the glass-ceramics depend on the aging process and material type.

Association between Dental Diseases and Oral Hygiene Care and the Risk of Dementia: A Retrospective Cohort Study.

OBJECTIVES: To investigate the associations of dental diseases and oral hygiene care with the risk of dementia. DESIGN: Retrospective longitudinal cohort study. SETTING AND PARTICIPANTS: We conducted a population-based study of individuals in the Korean National Health Insurance System. A total of 2,555,618 participants who underwent cardiovascular and dental screenings in 2008 were included. METHODS: Dental diseases including periodontal diseases, dental caries, and tooth loss were assessed by dentists. Information on oral hygiene care, including professional dental cleaning and the frequency of tooth brushing, was collected using a self-administered questionnaire. Study outcomes were all-cause dementia, Alzheimer's disease (AD), and vascular dementia (VaD). RESULTS: Periodontal diseases [adjusted hazard ratio (aHR) 1.07, 95% CI 1.04-1.09], dental caries (aHR 1.03, 95% CI 1.00-1.05), and 8-14 missing teeth (aHR 1.07, 95% CI 1.02-1.12) were associated with an increased risk of all-cause dementia. In contrast, either professional dental cleaning or frequent tooth brushing were associated with decreased risks of all-cause dementia (aHR 0.91, 95% CI 0.89-0.93 each; aHR 0.83, 95% CI 0.80-0.86 for both). The increased risks by dental diseases were reduced by oral hygiene care: periodontal diseases with professional dental cleaning (aHR 0.94, 95% CI 0.91-0.98) or tooth brushing ≥2 times/day (aHR 0.97, 95% CI 0.94-1.00) and 1 to 7 missing teeth with professional dental cleaning (aHR 0.94, 95% CI 0.89-0.98) or tooth brushing ≥2 times/day (aHR 0.92, 95% CI 0.89-0.95). Consistent results were noted for AD and VaD and in various subgroup analyses. CONCLUSION AND IMPLICATIONS: Periodontal disease, dental caries, and a high number of missing teeth were independently associated with a higher risk of dementia. Conversely, improved oral hygiene care, such as professional dental cleaning and frequent tooth brushing, may modify the risk of dementia associated with dental diseases.

Association of Dental Diseases and Oral Hygiene Care With the Risk of Heart Failure in Patients With Type 2 Diabetes: A Nationwide Cohort Study.

Background To evaluate the association of dental diseases and oral hygiene care with incidence of heart failure (HF) among patients with type 2 diabetes. Methods and Results This study included 173 927 patients with type 2 diabetes aged ≥40 years, who underwent Korean National Health Insurance Service health examinations in 2008 and were followed up until the end of 2017. Hazard ratios (HRs) and 95% CIs for HF were estimated using multivariable Cox proportional hazards regression analysis. During a median follow-up of 9.3 years, 1.94% of participants developed HF. An increased number of missing teeth was associated with a higher risk of HF (P<0.001). HRs of HF increased among individuals with ≥15 missing teeth (HR, 1.37 [95% CI, 1.14-1.64]) compared with those without missing teeth. Decreased risk of HF was observed in individuals with ≥1 time/year of professional dental cleaning (HR, 0.93 [95% CI, 0.87-0.99]) and in those with ≥2 times/d of toothbrushing (HR, 0.90 [95% CI, 0.82-0.98]) compared with those without these practices. While combined presence of missing teeth and periodontal disease (P=0.004) or dental caries (P=0.007) increased HF risk, combined oral hygiene care was associated with further HF risk reduction (P=0.024). Better oral hygiene care was associated with decreased HF risk, even as the number of missing teeth increased (P<0.001). Conclusions Among patients with type 2 diabetes, dental diseases and oral hygiene care are important determinants of HF development. Dental disease management and good oral care may prevent HF in patients with type 2 diabetes.

Effect of hemispherical dimples at titanium implant abutments for the retention of cemented crowns.

PURPOSE: The aim of this study was to assess the effect of hemispherical dimple structures on the retention of cobalt-chromium (Co-Cr) crowns cemented to titanium abutments, with different heights and numbers of dimples on the axial walls. MATERIALS AND METHODS: 3.0-mm and 6.0-mm abutments (N = 180) and Co-Cr crowns were prepared. The experimental groups were divided into two and four dimple groups. The crowns were cemented by TempBond and PANAVIA F 2.0 cements. The retention forces were measured after thermal treatments. A two-way Analysis of Variance (ANOVA) and post-hoc Tukey HSD test were conducted to analyze change in retention forces by use of dimples between groups, as well as t test for the effect of abutment height change (α = .05). RESULTS: Results of the two-way ANOVA showed a statistically significant difference in retention force due to the use of dimples, regardless of the types of cements used (P < .001). A significantly higher mean retention forces were observed in the groups with dimples than in the control group, using the post hoc Tukey HSD test (P < .001). Results of t test displayed a statistically significant increase in the retention force with 6.0-mm abutments compared with 3.0-mm abutments (P < .001). The groups without dimples revealed adhesive failure of cements, while the groups with dimples showed mixed failure of cements. CONCLUSION: Use of hemispherical dimples was effective for increasing retention forces of cemented crowns.

The comparison of the accuracy of temporary crowns fabricated with several 3D printers and a milling machine.

PURPOSE: The purpose of this in vitro study was to compare the accuracy of various 3D printers and a milling machine. MATERIALS AND METHODS: The die model was designed using CAD (Autodesk Inventor 2018 sp3). The 30 µm cement space was given to the die and the ideal crown of the mandibular left first molar was designed using CAD (ExoCAD). The crowns were produced using the milling machine (Imes-icore 250i) and the 3D printers (Zenith U, Zenith D, W11) and they were divided into four groups. In all groups, the interior of each crown was scanned (Identica blue) and superimposed (Geomagic Control X) with the previously designed die. The difference between the die and the actual crown was measured at specific points. The Kruskal-Wallis test, the Mann-Whitney test, and Bonferroni's method were performed with a statistical analysis software (P < .008 in inter-group comparison P < .001 in intra-group comparison). RESULTS: In all groups, the center of the occlusal area and the anti-rotational dimple area showed significantly greater difference and the marginal area showed the smallest difference comparatively. The mean value of the difference in each area and the sum of the differences were higher in order of W11, Imes-icore 250i, Zenith D, and Zenith U. CONCLUSION: The digital light processing (DLP) method shows higher accuracy compared to the sereolithography (SLA) method using the same resin material.

Chordin-like 1, a Novel Adipokine, Markedly Promotes Adipogenesis and Lipid Accumulation.

White adipose tissue serves as a metabolically dynamic organ that can synthesize and secrete biologically active compounds such as adipokines as well as a caloric reservoir for maintaining energy homeostasis. Adipokines are involved in diverse biological and physiological processes and there have been extensive attempts to characterize the effects of over two dozen adipokines. However, many of these adipokines are produced by not only adipose tissue, but also other tissues. Therefore, investigations into the effects of adipokines on physiological functions have been challenged. In this regard, we aimed to identify a new secreted protein that is encoded by genes specifically expressed in white adipose tissue through analysis of multi-tissue transcriptome and protein expression. As a result, we report a novel adipokine that is encoded by the adipose-specific gene, chordin-like 1 (Chrdl1), which is specifically expressed in white adipose tissue in mice; this expression pattern was conserved in the human orthologous CHRDL1 gene. The expression of Chrdl1 was enriched in fat cells and developmentally regulated in vitro and in vivo, and moreover, its retrovirus-mediated overexpression and recombinant protein treatment led to markedly increased adipogenesis. Further pathway enrichment analysis revealed enriched pathways related to lipogenesis and adipogenic signaling. Our findings support a pro-adipogenic role of CHRDL1 as a new adipokine and pave the way toward animal studies and future research on its clinical implications and development of anti-obesity therapy.

Influence of thermo-mechanical aging on fracture resistance and wear of digitally standardized chairside computer-aided-designed/computer-assisted-manufactured restorations.

OBJECTIVES: To investigate the influence of thermal cycling and mechanical loading (TCML) aging on fracture resistance and wear behavior of various chairside computer-aided-designed/computer-assisted-manufactured (CAD/CAM) premolar crowns cemented on standardized tooth abutments. METHODS: Eighty chairside CAD/CAM crowns were prepared using lithium disilicate (IPS e.max CAD; EM), zirconia-infiltrated lithium silicate (Celtra Duo; CD), polymer-infiltrated ceramic network (Vita Enamic; VE), and resin nanoceramics (Cerasmart; CS) (n = 20). The specimens were divided into two groups (n = 10). In one group, they were subjected to TCML: thermocycling (6000 cycles in distilled water at 5-55 °C) and mechanical loading (50 N for 1.2 × 106 cycles), while in control group they were stored in distilled water (37 °C for 24 h). The fracture load, height loss, and volume wear of the crowns were measured after TCML. Fractography was performed on fractured specimens. Data were analyzed using analysis of variance and multiple comparison tests (α=0.05). RESULTS: The mean fracture loads of EM and CD were significantly higher than those of EC and CS (p<0.05). There was no significant change in the fracture load of any CAD/CAM crowns after TCML (p>0.05). CS exhibited a significantly higher volume wear than the other materials investigated. The wear tracts of all TCML crowns acted as failure origins during the fracture test. CONCLUSIONS: The fracture resistance of glass-ceramic CAD/CAM crowns was significantly higher than that of resin composite crowns. A 5-year TCML aging did not affect the fracture resistance of CAD/CAM crowns investigated. However, TCML treatment produces a larger wear track in CS than in other materials. CLINICAL SIGNIFICANCE: Appropriate chairside CAD/CAM restorative material should be selected for successful clinical practice after considering the fracture and wear resistance of the crowns.

Synergistic effects of arginine-glycine-aspartic acid and phosphatidylserine on the surface immunomodulation and osseointegration of titanium implants.

The control of macrophage polarization is important in bone tissue regeneration such as osseointegration. In this study, a coating method was developed to improve the osseointegration of titanium (Ti) implants by generating an immunomodulatory effect. The surface of the Ti discs was coated with a poly(lactide-co-glycolide)(PLGA) polymer, phosphatidylserine (PS), and arginine-glycine-aspartic acid (RGD) peptide conjugated phospholipid. In in vitro assay using mouse bone marrow-derived macrophages (BMDMs), the most significant expression of the M2 marker genes (Arg-1, YM-1, FIZZ1) and CD206, an M2 surface marker, was obtained with coatings containing 6 mol% RGD conjugates and phospholipids consisting of 50 mol% PS. The M2-inducing effect of RGD and PS was also verified in rat femurs where coated Ti rods were implanted. The RGD and PS coating significantly enhanced the osseointegration of the Ti implants. Moreover, a biomechanical push-out test showed that the RGD and PS coating increased the interfacial binding force between the bone and implants. These results indicate that PS and RGD can be applied to the solid surface of implantable biomedical devices to improve immunomodulation and tissue regeneration.

Color and surface stainability of additively and subtractively manufactured interim restorative materials against mouth rinses.

STATEMENT OF PROBLEM: Interim restorations are often used along with mouth rinses during the healing period following surgical procedures. However, evidence regarding the color and surface properties of digitally fabricated interim restorations after oral rinsing is lacking. PURPOSE: The purpose of this in vitro study was to evaluate whether different mouth rinses could affect the color and surface roughness of milled and printed interim restorations after simulated oral rinsing. MATERIAL AND METHODS: Disk-shaped specimens (Ø15×2 mm; N=180) were fabricated by using conventional (Jet Tooth Shade), milled (Yamahachi PMMA Disk), and printed (NextDent C&B) resin materials. All resin specimens were divided into 3 different groups according to the rinsing material: distilled water, whitening mouth rinse (Listerine Healthy White), and conventional mouth rinse (Listerine Cool Mint). The specimens were further allocated into short- and long-term subgroups, and oral rinsing simulation was performed (n=10). Short-term rinsing simulated the conditions in a usual interim restoration period, and long-term rinsing was performed to evaluate the properties of the interim materials. The color differences (CIEDE2000, ΔE00) between the baseline and each time point were determined by using a spectrophotometer. The surface roughness of the tested specimens was measured by using a confocal laser scanning microscope. Kruskal-Wallis and Friedman tests with nonparametric pairwise comparisons were used to analyze the data (α=.05). RESULTS: On simulation of a 6-month use of the mouth rinse, the color change in the milled resin did not differ from that in the conventional resin (P>.334), but the printed resin showed a significantly greater color change than the other resins (P<.007). The greatest color change with the printed resin was observed when a conventional mouth rinse was used. However, all color changes were below a perceptible threshold of 1.30. When daily rinsing for 14 years was simulated, all resin groups showed a perceptible color change when conventional mouth rinse was used, and the printed resin showed the greatest median ±interquartile range ΔE00 (2.24 ±0.2). In both short- and long-term simulations, the printed resin rinsed with the conventional mouth rinse showed significantly greater roughness than that rinsed with distilled water (P<.009). CONCLUSIONS: The printed resin showed higher stainability than the conventional resin, and the color change was greatest with the conventional mouth rinse. However, in 6 months of daily mouth rinse simulation, all the tested resin materials exhibited imperceptible color change and clinically acceptable surface roughness.

A novel testis-enriched gene, Samd4a, regulates spermatogenesis as a spermatid-specific factor.

Spermatogenesis is the highly orchestrated process involving expression of a series of testicular genes. Testis-enriched genes are critical for cellular processes during spermatogenesis whose disruption leads to impaired spermatogenesis and male infertility. Nevertheless, among poorly investigated testicular genes are the mouse Samd4a and human SAMD4A which were identified in the current study as novel testis-enriched genes through transcriptomic analyses. In particular, as orthologous alternative splicing isoforms, mouse Samd4a E-form and human SAMD4AC-form containing the SAM domain were specific to testes. Western blot analyses revealed that the murine SAMD4AE-form was predominantly found in the testis. Analyses on GEO2R and single-cell RNA-seq datasets revealed that the Samd4a/SAMD4A expression was enriched in spermatids among various types of cells in adult testes. To investigate in vivo functions of Samd4a, Samd4a knockout mice were generated using the CRISPR/Cas9 system. The Samd4a deficiency resulted in lower testis weight, absence of elongated spermatids, and an increased number of apoptotic cells. Profiling of gene expression in human testis samples revealed that the SAMD4A expression was comparable between obstructive azoospermia patients and normal controls, but significantly lowered in nonobstructive azoospermia (NOA) patients. Among three subgroups of NOA, pre-meiotic arrest (NOA-pre), meiotic arrest (NOA-mei), and post-meiotic arrest (NOA-post), expression level of SAMD4A was higher in the NOA-post than the NOA-mei, but there was no difference between the NOA-pre and NOA-mei. The current studies demonstrated spermatid stage-specific expression of Samd4a/SAMD4A, and impairment of the late stages of spermatogenesis by disruption of the mouse Samd4a gene. These data suggest that Samd4a/SAMD4A plays an essential role in normal spermatogenesis, and SAMD4A, as a spermatid specific marker, can be used for subcategorizing NOA patients. Further understanding the molecular role of SAMD4A will advance our knowledge on genetic regulations in male infertility.

Evaluation of regeneration after the application of 2 types of deproteinized bovine bone mineral to alveolar bone defects in adult dogs.

PURPOSE: The aim of this study was to evaluate the preclinical results of 2 types of commercially available deproteinized bovine bone mineral (DBBM) when applied to alveolar bone defects in dogs. METHODS: This study was conducted using 6 beagles. Alveolar defects in the mandible were formed and filled with 2 DBBMs produced by a similar procedure. Defects were randomly assigned to be filled using DBBM 1 or 2. All defects were covered with a collagen membrane and had a healing period of 12 weeks. After the dogs were sacrificed, histological, histomorphometric, and linear/volumetric analyses were performed. RESULTS: Both DBBM groups showed similar histological findings, demonstrating that bone remodeling had occurred and new bone had formed. The residual bone particles were surrounded by newly formed vital bone. In the histomorphometric analysis, the ratio of the area of vital bone and residual bone substitute in DBBM 2 (38.18% and 3.47%, respectively) was higher than that of DBBM 1 (33.74% and 3.41%, respectively), although the difference was not statistically significant. There were also no statistically significant differences between both groups in linear and volumetric analyses using micro-computed tomography scans and digitized images of dental casts. CONCLUSIONS: In the present study, DBBM 1and 2, which were produced by similar processes, showed similar results in histological, histomorphometric, and volumetric analyses. Further studies are needed to identify more specific differences between the 2 DBBMs.

Loss of Monoallelic Expression of IGF2 in the Adult Liver Via Alternative Promoter Usage and Chromatin Reorganization.

In mammals, genomic imprinting operates via gene silencing mechanisms. Although conservation of the imprinting mechanism at the H19/IGF2 locus has been generally described in pigs, tissue-specific imprinting at the transcript level, monoallelic-to-biallelic conversion, and spatio-temporal chromatin reorganization remain largely uninvestigated. Here, we delineate spatially regulated imprinting of IGF2 transcripts, age-dependent hepatic mono- to biallelic conversion, and reorganization of topologically associating domains at the porcine H19/IGF2 locus for better translation to human and animal research. Whole-genome bisulfite sequencing (WGBS) and RNA sequencing (RNA-seq) of normal and parthenogenetic porcine embryos revealed the paternally hypermethylated H19 differentially methylated region and paternal expression of IGF2. Using a polymorphism-based approach and omics datasets from chromatin immunoprecipitation sequencing (ChIP-seq), whole-genome sequencing (WGS), RNA-seq, and Hi-C, regulation of IGF2 during development was analyzed. Regulatory elements in the liver were distinguished from those in the muscle where the porcine IGF2 transcript was monoallelically expressed. The IGF2 transcript from the liver was biallelically expressed at later developmental stages in both pigs and humans. Chromatin interaction was less frequent in the adult liver compared to the fetal liver and skeletal muscle. The duration of genomic imprinting effects within the H19/IGF2 locus might be reduced in the liver with biallelic conversion through alternative promoter usage and chromatin remodeling. Our integrative omics analyses of genome, epigenome, and transcriptome provided a comprehensive view of imprinting status at the H19/IGF2 cluster.