Identification of Type-H-like Blood Vessels in a Dynamic and Controlled Model of Osteogenesis in Rabbit Calvarium.

Angiogenesis and bone regeneration are closely interconnected processes. Whereas type-H blood vessels are abundantly found in the osteogenic zones during endochondral long bone development, their presence in flat bones' development involving intramembranous mechanisms remains unclear. Here, we hypothesized that type-H-like capillaries that highly express CD31 and Endomucin (EMCN), may be present at sites of intramembranous bone development and participate in the control of osteogenesis. A rabbit model of calvarial bone augmentation was used in which bone growth was controlled over time (2-4 weeks) using a particulate bone scaffold. The model allowed the visualization of the entire spectrum of stages throughout bone growth in the same sample, i.e., active ossification, osteogenic activity, and controlled inflammation. Using systematic mRNA hybridization, the formation of capillaries subpopulations (CD31-EMCN staining) over time was studied and correlated with the presence of osteogenic precursors (Osterix staining). Type-H-like capillaries strongly expressing CD31 and EMCN were identified and described. Their presence increased gradually from the regenerative zone up to the osteogenic zone, at 2 and 4 weeks. Type-H-like capillaries may thus represent the initial vascular support encountered in flat bones' development and which organize osteogenic niches.

Influence of the Titanium Base Abutment Design on Monolithic Zirconia Multiple-Unit Implant Fixed Dental Prostheses: A Laboratory Study.

PURPOSE: This in vitro study evaluated technical outcomes of monolithic zirconia implant-supported fixed dental prostheses (iFDPs) supported by different designs of titanium base abutments after aging and static load testing. MATERIALS AND METHODS: Sixty three-unit monolithic zirconia (yttrium oxide partially stabilized tetragonal zirconia polycrystals; Y-TZP)iFDPs were produced and divided into four groups: group A-conical titanium base abutments for the prosthesis; group B-cylindrical titanium base abutments for the crown; group C-conical titanium base abutment for the prosthesis and cylindrical titanium base abutment for the crown; group D-cylindrical titanium base abutments for the prosthesis. The samples were subjected to an aging protocol and to continuous static loading until failure and analyzed visually and with specific software. The technical outcomes comprised the occurrence of debonding, screw loosening, deformation and fracture, abutment deformation and fracture, implant deformation and fracture, zirconia chipping and fracture, and bending moments. The Pearson chi-squared test (χ2) and Fischer exact test were used to compare the outcomes. The Kolmogorov-Smirnov test was used to evaluate data distribution of the bending moments. Analysis of variance (ANOVA) was used for the analysis of parametric data distribution, and the Kruskal-Wallis test was used for nonparametric data distribution. RESULTS: After aging, a higher percentage of debonding (P = .042) and micromovement (P = .034) was recorded in group C (P = .042). The conical titanium base abutments had a higher debonding (P = .049) and a higher macromovement rate (P = .05). The static load test showed higher bending moments in group D (P = .001) and lower bending moments in group A (P = .001). Debonding was highest in group C (P = .001) and lowest in group A (P = .002). Complete loss of retention rate was highest in group C (P = .001). The conical titanium base abutment had the highest debonding rate (P = .001) and complete loss of retention (P = .001). The micromovement rate was the highest for cylindrical titanium base abutments for the crown (P = .001). The lowest screw loosening, zirconia chipping, and fracture rate (P = .001) and the highest screw deformation (P = .004) were recorded in group A. The screw deformation rate was lowest in the cylindrical titanium base abutments for the crown (P = .008). CONCLUSION: The mixed titanium base abutment design comprising one conical and one cylindrical abutment in Y-TZP iFDPs led to the highest debonding rate. The cylindrical titanium base abutment for the prosthesis provided a lower percentage of debonding and the highest load resistance.

Effect of Multiple Enamel Surface Treatments on Micro-Shear Bond Strength.

Objectives: The aim of this study was to evaluate the effect of different enamel surface treatments on the micro-shear bond strength. Materials and methods: Sixty-four approximal surfaces from freshly extracted molars were randomly assigned to eight groups, according to combinations of the following enamel surface treatments: ground or unground, 37.5% phosphoric acid etching time of 15 or 30 s, and with or without primer application. The same bonding agent (Optibond FL™ Adhesive, Kerr) was then used for all groups, and a 1.8 mm diameter resin composite (Harmonize™, Kerr) cylinder was built up on the bonded surface. Samples underwent a shear force test at a crosshead speed of 1 mm/min until failure. Bond strength was calculated, and failure modes were inspected under an optical microscope. Results: Bond-strength values ranged from 8.2 MPa for 15 s etched unground enamel with primer application to 19.6 MPa for 30 s etched ground enamel without primer application. ANOVA and Fisher's LSD post hoc tests revealed significant differences between the groups. Conclusions: Etching time and grinding have a statistically significant effect on the micro-shear bond strength of a three-step etch-and-rinse adhesive system on enamel. Primer application does not seem to be beneficial for enamel adhesion.

CAD-CAM complete denture resins: an evaluation of biocompatibility, mechanical properties, and surface characteristics.

OBJECTIVES: This study evaluated the biocompatibility, mechanical properties, and surface roughness of CAD-CAM milled and rapidly-prototyped/3D-printed resins used for manufacturing complete dentures. METHODS: Six groups of resin specimens were prepared, milled-base (MB), milled-tooth shade (MT), printed-tooth shade (PT), printed-base with manufacturer-recommended 3D-printer (PB1), printed-base with third-party 3D-printer (PB2), printed-base in a vertical orientation (PB2V). Human epithelial (A-431) and gingival (HGF-1) cells were cultured and tested for biocompatibility using Resazurin assays. Three-point bending and nanoindentation tests measured the mechanical properties of the resin groups. Surface roughness was evaluated using a high-resolution laser profilometer. ANOVA and post-hoc tests were used for statistical analyses (α = 0.05). RESULTS: There were no significant differences in biocompatibility between any of the investigated groups. MB revealed a higher ultimate strength (p = 0.008), elastic modulus (p = 0.002), and toughness (p = 0.014) than PB1. MT had significantly higher elastic modulus than PT (p < 0.001). Rapidly-prototyped resin samples with a manufacturer-recommended 3D-printer (PB1) demonstrated higher ultimate strength (p = 0.008), elastic modulus (p < 0.001), hardness (p < 0.001) and a reduced surface roughness (p < 0.05) when compared with rapidly-prototyped groups using a third-party 3D-printer (PB2). Rapidly-prototyped samples manufactured with a vertical printing orientation (PB2V) revealed a significantly lower elastic modulus than samples groups manufactured using horizontal printing orientation (PB2) (p = 0.011). CONCLUSIONS: Within the limits of this present study, CAD-CAM milled and rapidly-prototyped complete denture resins performed similarly in terms of biocompatibility and surface roughness. However, the milled denture resins were superior to the rapidly-prototyped denture resins with regard to their mechanical properties. Printing orientation and type of 3D-printer can affect the resin strength and surface roughness.

Validation of a novel diagnostic tool for decreased tongue pressure.

INTRODUCTION: Reduced tongue pressure may render eating and swallowing difficult. The purpose of this study was to investigate whether the tongue training device can also be used as a diagnostic device and whether its sensitivity and specificity are equal to the numerical tongue pressure measuring device. MATERIAL AND METHODS: The target group is patients aged 70 years and over who are hospitalised for rehabilitation. Tongue pressure was measured by both, a tongue pressure measuring instrument and a tongue training tool. The diagnosis of the reduced tongue pressure was made with the tongue pressure measuring instrument and set the verified with the novel tongue training tool. RESULTS: Sixty-two participants were included in the study. Forty-five were classified by the tongue pressure measuring device and 53 by the tongue training device as 'low tongue pressure'. Spearman correlation confirmed a positive correlation between the tongue pressure measuring device and the tongue training device rs  = 0.800, p = 0.01 level (2-tailed). The tongue training device test identified sensitivity was 100%, and its specificity was 52.9%. The AUC of the ROC curve is 0.901. CONCLUSION: The tongue training device seems a simple, safe and readily available alternative to the tongue pressure measuring device for the diagnosis of low tongue pressure, with an excellent sensitivity and very good specificity.

Pre-Treat Xenogenic Collagenous Blocks of Bone Substitutes with Saline Facilitate Their Manipulation and Guarantee High Bone Regeneration Rates, Qualitatively and Quantitatively.

Deproteinized bovine bone mineral particles embedded in collagen (DBBM-C) are widely used for bone regenerations with excellent, albeit sometimes variable clinical outcomes. Clinicians usually prepare DBBM-C by mixing with blood. Replacing blood by saline represents an alternative. We investigated if saline treatment could improve DBBM-C i. handling in vitro and ii. biological performances in a rabbit calvarial model. In vitro, DBBM-C blocks soaked in saline or blood were submitted to compression tests. In vivo, four poly ether ether ketone (PEEK)cylinders were placed on 16 rabbit skulls, filled with DBBM-C soaked in blood or saline for 2-4-8-12 weeks before histomorphometry. DBBM-C blocks were fully hydrated after 30 s in saline when 120 s in blood could not hydrate blocks core. Stiffness gradually decreased 2.5-fold after blood soaking whereas a six-fold decrease was measured after 30 s in saline. In vivo, saline treatment allowed 50% more bone regeneration during the first month when compared to blood soaking. This difference was then no longer visible. New bone morphology and maturity were equivalent in both conditions. DBBM-C saline-soaking facilitated its handling and accelerated bone regeneration of highly qualitative tissues when compared to blood treatment. Saline pretreatment thus may increase the clinical predictability of bone augmentation procedures.

Effect of airborne-particle abrasion of a titanium base abutment on the stability of the bonded interface and retention forces of crowns after artificial aging.

STATEMENT OF PROBLEM: Published data have shown that a mechanical surface treatment of titanium surfaces increases bonding potential. However, most of the studies are based on shear or tensile tests performed on flat-surfaced specimens and do not take into consideration the retention given by the titanium base (ti-base) axial walls and the thermomechanical loading seen in a clinical setting. PURPOSE: The purpose of this in vitro study was to evaluate the influence of different airborne-particle abrasion (APA) methods of the ti-base surface on the stability of the bonded interface and retention forces between these titanium bases and lithium disilicate crowns after thermomechanical aging. MATERIAL AND METHODS: Sixty internal connection implants (Conelog) were restored with lithium disilicate crowns and bonded to the corresponding ti-bases (Conelog). The ti-bases were divided into 4 groups (n=15), 3 experimental groups applying different APA methods, 30-µm silica-modified Al2O3 particles (CoJet) (30-SiO-AlO), 50-µm Al2O3 (Cobra Aluoxyd) (50-AlO), 110-µm silica-modified Al2O3 particles (Rocatec Plus) (110-SiO-AlO), and 1 control group (NoT). Ti-bases were airborne-particle abraded (10 seconds, 0.25 MPa at a 10-mm distance) under standardized conditions in a custom-made APA device. All crowns were cemented with a resin cement (Multilink Hybrid Abutment). After aging (1 200 000 cycles, 49 N, 1.67 Hz; 5 °C-55 °C, 120 seconds), all specimens were assessed for the presence of bond failures by optical microscopy (×50). The retention forces (N) were tested by using a pull-off test (0.5mm/min). Modes of failure were classified (Type 1, 2, or 3). An additional ti-base representing each group was prepared for surface roughness (µm) calculation (Ra, Rc, Rz) with a noncontact laser profilometer, and representative scanning electron microscope (SEM) images were recorded (×1000). Chi-squared tests were performed to analyze the bonded interface failure and modes of failure, and a Kruskal-Wallis test was selected to evaluate retention force values (α=.05). RESULTS: The percentages of bonding failure after aging were 73.3% (NoT), 40% (30-SiO-AlO), 6.7% (50-AlO), and 40% (110-SiO-AlO). The stability of the bonded interface was influenced by the APA method applied (P<.05). Mean ±standard deviation retention force values varied from 206.3 ±86.3 N (NoT) to 420 ±139.5 N (50-AlO), and the differences between these 2 groups were significant (P<.05). Modes of failure were predominantly Type 2 (30-SiO-AlO; 50-AlO; 110-SiO-AlO) and Type 3 (NoT). CONCLUSIONS: Airborne-particle abrasion of the titanium surface increased the bond stability and retention forces between the ti-base and the respective crown. The use of 50-µm Al2O3 provided the most stable bonded interface among the different treatments.

Effect of a Modified Irrigation Protocol on the Cleanliness of Moderately Curved Canals.

OBJECTIVES: This study tested the hypothesis that modifying the sequence of sodium hypochlorite (NaOCl)/ethylene diamine tetra-acetic acid (EDTA) irrigation during root canal shaping would improve apical cleanliness in moderately curved canals. MATERIALS AND METHODS: Forty-five root canals were prepared using Protaper Gold with three irrigation protocols. Standard irrigation (SI) used 0.5 mL 3% NaOCl between each instrument, followed by 5 mL 17% EDTA manually agitated for 30 seconds. Reverse irrigation (RI) used 0.5 mL of 17% EDTA between each instrument, then 5 mL of 3% NaOCl, manually agitated for 30 seconds. Reverse irrigation plus (RI+) was similar to RI, except NaOCl (5 mL), used as a final rinse, was allowed to interact for 3 minutes with dentin before manual agitation (30 seconds).Root canal cleanliness was evaluated under the scanning electron microscope (SEM) (Hulsmann score); the chemical composition of dentin after irrigation was analyzed by energy dispersive X-ray (EDX). STATISTICAL ANALYSIS: Results were compared using Kruskal-Wallis ANOVA by ranks and Wilcoxon matched paired posthoc tests. A Chi-square test assessed whether the best cleanliness would demonstrate a significant association with one irrigation protocol; odds ratio calculation was performed using score "1" versus score "2 or more" (2+) (p < 0.05). RESULTS: In the apical region, cleanliness was better in RI+ than SI and both significantly better than RI. Odd ratios indicate that the cleanliness in RI+ was significantly better than RI and SI groups (p < 0.000 and 0.003, respectively). Independently of the irrigation protocol, EDX analyses showed no chemical alteration of root dentin. CONCLUSIONS: Using 17% EDTA during shaping, followed by 3% NaOCl rinse for 3 minutes, improved apical cleanliness without inducing erosion of dentin.

Calvarial Model of Bone Augmentation in Rabbit for Assessment of Bone Growth and Neovascularization in Bone Substitution Materials.

The basic principle of the rabbit calvarial model is to grow new bone tissue vertically on top of the cortical part of the skull. This model allows assessment of bone substitution materials for oral and craniofacial bone regeneration in terms of bone growth and neovascularization support. Once animals are anesthetized and ventilated (endotracheal intubation), four cylinders made of polyether ether ketone (PEEK) are screwed onto the skull, on both sides of the median and coronal sutures. Five intramedullary holes are drilled within the bone area delimited by each cylinder, allowing influx of bone marrow cells. The material samples are placed into the cylinders which are then closed. Finally, the surgical site is sutured, and animals are awaken. Bone growth may be assessed on live animals by using microtomography. Once animals are euthanized, bone growth and neovascularization may be evaluated by using microtomography, immune-histology and immunofluorescence. As the evaluation of a material requires maximum standardization and calibration, the calvarial model appears ideal. Access is very easy, calibration and standardization are facilitated by the use of defined cylinders and four samples may be assessed simultaneously. Furthermore, live tomography may be used and ultimately a large decrease in animals to be euthanized may be anticipated.

Translational research on clinically failed zirconia implants.

OBJECTIVES: To provide fractographic analysis of clinically fractured zirconia implants recovered with their cemented crown. To calculate bending moments, corresponding stress and crack onset location on the implant's fracture surface using a mathematical model integrating spatial coordinates of the crown-implant part and occlusal loading obtained from 2D and 3D images. METHODS: 15 fractured zirconia implants parts (11 posterior and 4 anterior) with their all- ceramic crowns still cemented on it were recovered. The implants were first generations from four manufacturers (AXIS Biodental, Z-Systems, Straumann, Swiss Dental Solutions). The time-to-failure varied between 2weeks and 9years. Fractography was performed identifying the failure origin and characteristic surface crack features. From 2D and 3D digital images of the crown-implant part, spatial coordinates anchoring the crown's occlusal contacts with the implant's central axis and reference plane were integrated in a mathematical model spreadsheet. Loads of 500 N in total were selectively distributed over identified occlusal contacts from wear patterns. The resultant bending and torsion moments, corresponding shear, tensile, maximum principal stress and von Mises stress were calculated. The fracture crack onset location on the implant's fracture surface was given by an angular position with respect to an occlusal reference and compared with the location of the fracture origin identified from fractographic analysis. RESULTS: Implants fractured from the periphery of the smaller inner diameter between two threads at the bone-entrance level except for one implant which failed half-way within the bone. The porous coating (AXIS Biodental) and the large grit alumina sandblasting (Z-System) created surface defects directly related to the fracture origin. The model spreadsheet showed how occlusal loading with respect to the implant's central axis affects bending moments and crack onset. Dominant loads distributed on contacts with important wear pattern provided a calculated crack onset location in good agreement with the fractographic findings of the fracture origin. SIGNIFICANCE: Recovered broken zirconia implant parts with their restorative crowns can provide not only information regarding the failure origin using fractography but also knowledge regarding occlusal crown loading with respect to the implant's axis. The mathematical model was helpful in showing how occlusal loading affects the location of the fracture initiation site on clinical zirconia implant fracture cases.