Protocol to decellularize porcine right ventricular outflow tracts using a 3D printed flow chamber.

Surgical treatment of pediatric congenital heart disease with tissue grafts is a lifesaving intervention. Decellularization to reduce immunogenicity of tissue grafts is an increasingly popular alternative to glutaraldehyde fixation. Here, we present a protocol to decellularize porcine right ventricular outflow tracts using a 3D printed flow chamber. We describe steps for 3D printing the flow rig, preparing porcine tissue, and using the flow rig to utilize shear forces for decellularization. We then detail procedures for characterizing the acellular scaffold. For complete details on the use and execution of this protocol, please refer to Vafaee et al.1.

The Role of Immersive Virtual Reality and Augmented Reality in Medical Communication: A Scoping Review.

Communication between clinicians and patients and communication within clinical teams is widely recognized as a tool through which improved patient outcomes can be achieved. As emerging technologies, there is a notable lack of commentary on the role of immersive virtual reality (VR) and augmented reality (AR) in enhancing medical communication. This scoping review aims to map the current landscape of literature on this topic and highlights gaps in the evidence to inform future endeavors. A comprehensive search strategy was conducted across 3 databases (PubMed, Web of Science, and Embase), yielding 1000 articles, of which 623 were individually screened for relevance. Ultimately, 22 articles were selected for inclusion and review. Similarities across the cohort of studies included small sample sizes, observational study design, use of questionnaires, and more VR studies than AR. The majority of studies found these technologies to improve medical communication, although user tolerability limitations were identified. More studies are required, presenting more robust findings, in order to draw more definitive conclusions and stronger recommendations for use of immersive VR/AR in clinical environments.

Biological Scaffolds for Congenital Heart Disease.

Congenital heart disease (CHD) is the most predominant birth defect and can require several invasive surgeries throughout childhood. The absence of materials with growth and remodelling potential is a limitation of currently used prosthetics in cardiovascular surgery, as well as their susceptibility to calcification. The field of tissue engineering has emerged as a regenerative medicine approach aiming to develop durable scaffolds possessing the ability to grow and remodel upon implantation into the defective hearts of babies and children with CHD. Though tissue engineering has produced several synthetic scaffolds, most of them failed to be successfully translated in this life-endangering clinical scenario, and currently, biological scaffolds are the most extensively used. This review aims to thoroughly summarise the existing biological scaffolds for the treatment of paediatric CHD, categorised as homografts and xenografts, and present the preclinical and clinical studies. Fixation as well as techniques of decellularisation will be reported, highlighting the importance of these approaches for the successful implantation of biological scaffolds that avoid prosthetic rejection. Additionally, cardiac scaffolds for paediatric CHD can be implanted as acellular prostheses, or recellularised before implantation, and cellularisation techniques will be extensively discussed.

Influence of Thickness of Opaque Porcelain and Alloy Color on the Final Color of Porcelain-Fused-to-Metal Restorations.

Despite the advent of metal-free solutions, porcelain-fused-to-metal restorations (PFM) are still widely used. Particularly for the latest ceramic systems, scarce information is present in the scientific literature about the ideal opaque layer thickness and the alloy color impact to achieve the desired final color of PFM restorations. This study aimed to evaluate the influence of opaque thickness variation layered on different metal alloys on the final color of PFM restorations. Opaque porcelain of one metal−ceramic system (VITA VM13) was layered in four different thicknesses (0.10 mm, 0.15 mm, 0.20 mm, and 0.25 mm) on three differently colored dental alloys: a gold−platinum alloy (yellowish), a gold−palladium alloy (light grayish), and a nickel−chromium alloy (dark grayish). The veneering porcelain layered over the opaque was kept constant (Base Dentine 0.45 mm, Transpa Dentine 0.50 mm, and Enamel 0.20 mm). Sixty specimens were fabricated, five samples for each combination of alloy/opaque thickness. The color difference (ΔE) between specimen and reference was measured using a clinical spectrophotometer. The two-way ANOVA revealed that the thickness of both the opaque (p < 0.001) and the metal alloy (p < 0.001) significantly influenced the ΔE values. Gray-colored alloys covered by a 0.10 mm thick opaque layer enabled the closest color match, whereas this occurred for yellow-color alloys covered by a 0.15 mm thick opaque layer. In contrast, the samples covered by a 0.25 mm thick opaque layer obtained the worst ΔE.

Cardiac pericyte reprogramming by MEK inhibition promotes arteriologenesis and angiogenesis of the ischemic heart.

Pericytes (PCs) are abundant yet remain the most enigmatic and ill-defined cell population in the heart. Here, we investigated whether PCs can be reprogrammed to aid neovascularization. Primary PCs from human and mouse hearts acquired cytoskeletal proteins typical of vascular smooth muscle cells (VSMCs) upon exclusion of EGF/bFGF, which signal through ERK1/2, or upon exposure to the MEK inhibitor PD0325901. Differentiated PCs became more proangiogenic, more responsive to vasoactive agents, and insensitive to chemoattractants. RNA sequencing revealed transcripts marking the PD0325901-induced transition into proangiogenic, stationary VSMC-like cells, including the unique expression of 2 angiogenesis-related markers, aquaporin 1 (AQP1) and cellular retinoic acid-binding protein 2 (CRABP2), which were further verified at the protein level. This enabled us to trace PCs during in vivo studies. In mice, implantation of Matrigel plugs containing human PCs plus PD0325901 promoted the formation of αSMA+ neovessels compared with PC only. Two-week oral administration of PD0325901 to mice increased the heart arteriolar density, total vascular area, arteriole coverage by PDGFRß+AQP1+CRABP2+ PCs, and myocardial perfusion. Short-duration PD0325901 treatment of mice after myocardial infarction enhanced the peri-infarct vascularization, reduced the scar, and improved systolic function. In conclusion, myocardial PCs have intrinsic plasticity that can be pharmacologically modulated to promote reparative vascularization of the ischemic heart.

Recent Advances in KEAP1/NRF2-Targeting Strategies by Phytochemical Antioxidants, Nanoparticles, and Biocompatible Scaffolds for the Treatment of Diabetic Cardiovascular Complications.

Significance: Modulation of nuclear factor (erythroid-derived 2)-like 2 (Nrf2)-mediated antioxidant response is a key aspect in the onset of diabetes-related cardiovascular complications. With this review, we provide an overview of the recent advances made in the development of Nrf2-targeting strategies for the treatment of diabetes, with particular attention toward the activation of Nrf2 by natural antioxidant compounds, nanoparticles, and oxidative stress-modulating biocompatible scaffolds. Recent Advances: In the past 30 years, studies addressing the use of antioxidant therapies to treat diabetes have grown exponentially, showing promising but yet inconclusive results. Animal studies and clinical trials on the Nrf2 pathway have shown promising results, suggesting that its activation can delay or reverse some of the cardiovascular impairments in diabetes. Critical Issues: Hyperglycemia- and oscillating glucose levels-induced reactive oxygen species (ROS) accumulation is progressively emerging as a central factor in the onset and progression of diabetes-related cardiovascular complications, including endothelial dysfunction, retinopathy, heart failure, stroke, critical limb ischemia, ulcers, and delayed wound healing. In this context, accumulating evidence suggests a central role for Nrf2-mediated antioxidant response, one of the most studied cellular defensive mechanisms against ROS accumulation. Future Directions: Innovative approaches such as tissue engineering and nanotechnology are converging toward targeting oxidative stress in diabetes. Antioxid. Redox Signal. 36, 707-728.

Cement opacity and color as influencing factors on the final shade of metal-free ceramic restorations.

OBJECTIVE: To investigate the influence of opacity and color of luting cements on the final shade of metal-free restorations. MATERIALS AND METHODS: Five resin cement colors in combination with four different thicknesses of CAD/CAM ceramic materials were tested, and a composite substrate was used as dentin color reference (n = 3). Specimen color was measured with a spectrophotometer equipped with an integrating sphere before and after cementation (CIELAB). Cement and ceramic color and opacity (TP) were assessed by measuring the tested ceramic thickness as a 1-mm thick disk for each of the cement shades. The differences in color were evaluated (ΔE00 ). Data were statistically analyzed by a Two-Way ANOVA followed by the Tukey Test for post-hoc comparison (P < .05) and multiple comparison Pearson's test (P < .05); the acceptability and perceptibility threshold were evaluated. RESULTS: Statistically significant influence was found for factors ceramic thickness and cement shade (P < .001). Perceptible and unacceptable color changes were induced on the final restoration by resin cements (ranging from ΔE00  = 0.69 ± 0.54 to ΔE00  = 5.53 ± 0.46), the correlation between factors became strong (r2 > 0.6) in case of mismatch between color and translucency of cement and ceramic. Only the clear shade in combination with the thickest ceramic, resulted in an imperceptible color change (ΔE00  = 0.69 ± 0.54). CONCLUSIONS: The final shade of ceramic restorations can be influenced by resin cements; the magnitude of influence is related to the cement optical properties. CLINICAL SIGNIFICANCE: In order to influence the final shade of a ceramic restoration, a cement more opaque than the restorative material should be used. Conversely, in the case of a fitting shade of the restoration, a cement more translucent than the restoration should be used to avoid undesired changes.

The SARS-CoV-2 Spike protein disrupts human cardiac pericytes function through CD147 receptor-mediated signalling: a potential non-infective mechanism of COVID-19 microvascular disease.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes a broad range of clinical responses including prominent microvascular damage. The capacity of SARS-CoV-2 to infect vascular cells is still debated. Additionally, the SARS-CoV-2 Spike (S) protein may act as a ligand to induce non-infective cellular stress. We tested this hypothesis in pericytes (PCs), which are reportedly reduced in the heart of patients with severe coronavirus disease-2019 (COVID-19). Here we newly show that the in vitro exposure of primary human cardiac PCs to the SARS-CoV-2 wildtype strain or the α and δ variants caused rare infection events. Exposure to the recombinant S protein alone elicited signalling and functional alterations, including: (1) increased migration, (2) reduced ability to support endothelial cell (EC) network formation on Matrigel, (3) secretion of pro-inflammatory molecules typically involved in the cytokine storm, and (4) production of pro-apoptotic factors causing EC death. Next, adopting a blocking strategy against the S protein receptors angiotensin-converting enzyme 2 (ACE2) and CD147, we discovered that the S protein stimulates the phosphorylation/activation of the extracellular signal-regulated kinase 1/2 (ERK1/2) through the CD147 receptor, but not ACE2, in PCs. The neutralisation of CD147, either using a blocking antibody or mRNA silencing, reduced ERK1/2 activation, and rescued PC function in the presence of the S protein. Immunoreactive S protein was detected in the peripheral blood of infected patients. In conclusion, our findings suggest that the S protein may prompt PC dysfunction, potentially contributing to microvascular injury. This mechanism may have clinical and therapeutic implications.

Comparison between Hydrofluoric Acid and Single-Component Primer as Conditioners on Resin Cement Adhesion to Lithium Silicate and Lithium Disilicate Glass Ceramics.

This study aimed at evaluating the effects of different surface conditionings on the microshear bond strength (µSBS) of a self-adhesive resin cement to VITA Suprinity (ZLS) and IPS e.max CAD (LD). Three surface conditioning protocols were performed on ZLS and LD before luting with a self-adhesive resin cement (RelyX Unicem 2, RXU): hydrofluoric acid (HF), HF + silane (HF + S), or Monobond Etch & Prime (EP). In each group, 15 cylindrical buildups of RXU were prepared on five milled bars and submitted to a µSBS test. Data were statistically analyzed with two-way ANOVA and Tukey's post hoc test (p < 0.05). Failure modes were recorded and classified as adhesive, mixed, cohesive in resin, or ceramic, and statistically analyzed with Fisher's exact test (p = 0.05). One additional bar per group was used for the morphological characterization of the conditioned surface by means of SEM. The material per se did not significantly influence adhesion (p = 0.744). Conditioning protocol was a significant factor: EP yielded significantly higher µSBS than HF (p = 0.005), while no significant differences emerged between EP and HF + S (p = 0.107), or HF + S and HF (p = 0.387). The material-conditioning protocol interaction was not statistically significant (p = 0.109). Significant intergroup differences were found in distribution of failure modes: mixed failures were predominant in the ZLS/EP group, while the other groups showed a prevalence of adhesive failures. The self-etching primer showed promising results in terms of immediate bond strength of a self-adhesive resin cement to lithium-silica-based glass ceramics, suggesting its alternative use to hydrofluoric acid and silane conditioning protocols.

Reconstruction of the Swine Pulmonary Artery Using a Graft Engineered With Syngeneic Cardiac Pericytes.

The neonatal heart represents an attractive source of regenerative cells. Here, we report the results of a randomized, controlled, investigator-blinded preclinical study, which assessed the safety and effectiveness of a matrix graft cellularized with cardiac pericytes (CPs) in a piglet model of pulmonary artery (PA) reconstruction. Within each of five trios formed by 4-week-old female littermate piglets, one element (the donor) was sacrificed to provide a source of CPs, while the other two elements (the graft recipients) were allowed to reach the age of 10 weeks. During this time interval, culture-expanded donor CPs were seeded onto swine small intestinal submucosa (SIS) grafts, which were then shaped into conduits and conditioned in a flow bioreactor. Control unseeded SIS conduits were subjected to the same procedure. Then, recipient piglets were randomized to surgical reconstruction of the left PA (LPA) with unseeded or CP-seeded SIS conduits. Doppler echocardiography and cardiac magnetic resonance imaging (CMRI) were performed at baseline and 4-months post-implantation. Vascular explants were examined using histology and immunohistochemistry. All animals completed the scheduled follow-up. No group difference was observed in baseline imaging data. The final Doppler assessment showed that the LPA's blood flow velocity was similar in the treatment groups. CMRI revealed a mismatch in the average growth of the grafted LPA and contralateral branch in both treatment groups. Histology of explanted arteries demonstrated that the CP-seeded grafts had a thicker luminal cell layer, more intraparietal arterioles, and a higher expression of endothelial nitric oxide synthase (eNOS) compared with unseeded grafts. Moreover, the LPA stump adjacent to the seeded graft contained more elastin and less collagen than the unseeded control. Syngeneic CP engineering did not accomplish the primary goal of supporting the graft's growth but was able to improve secondary outcomes, such as the luminal cellularization and intraparietal vascularization of the graft, and elastic remodeling of the recipient artery. The beneficial properties of neonatal CPs may be considered in future bioengineering applications aiming to reproduce the cellular composition of native arteries.

Human adventitial pericytes provide a unique source of anti-calcific cells for cardiac valve engineering: Role of microRNA-132-3p.

AIMS: Tissue engineering aims to improve the longevity of prosthetic heart valves. However, the optimal cell source has yet to be determined. This study aimed to establish a mechanistic rationale supporting the suitability of human adventitial pericytes (APCs). METHODS AND RESULTS: APCs were immunomagnetically sorted from saphenous vein leftovers of patients undergoing coronary artery bypass graft surgery and antigenically characterized for purity. Unlike bone marrow-derived mesenchymal stromal cells (BM-MSCs), APCs were resistant to calcification and delayed osteochondrogenic differentiation upon high phosphate (HP) induction, as assessed by cytochemistry and expression of osteogenic markers. Moreover, glycolysis was activated during osteogenic differentiation of BM-MSCs, whereas APCs showed no increase in glycolysis upon HP challenge. The microRNA-132-3p (miR-132), a known inhibitor of osteogenesis, was found constitutively expressed by APCs and upregulated following HP stimulation. The anti-calcific role of miR-132 was further corroborated by in silico analysis, luciferase assays in HEK293 cells, and transfecting APCs with miR-132 agomir and antagomir, followed by assessment of osteochondrogenic markers. Interestingly, treatment of swine cardiac valves with APC-derived conditioned medium conferred them with resistance to HP-induced osteogenesis, with this effect being negated when using the medium of miR-132-silenced APCs. Additionally, as an initial bioengineering step, APCs were successfully engrafted onto pericardium sheets, where they proliferated and promoted aortic endothelial cells attraction, a process mimicking valve endothelialization. CONCLUSIONS: Human APCs are resistant to calcification compared with BM-MSCs and convey the anti-calcific phenotype to heart valves through miR-132. These findings may open new important avenues for prosthetic valve cellularization.

Three-Dimensional Printable Enzymatically Active Plastics.

Here, we describe a facile route to the synthesis of enzymatically active highly fabricable plastics, where the enzyme is an intrinsic component of the material. This is facilitated by the formation of an electrostatically stabilized enzyme-polymer surfactant nanoconstruct, which, after lyophilization and melting, affords stable macromolecular dispersions in a wide range of organic solvents. A selection of plastics can then be co-dissolved in the dispersions, which provides a route to bespoke 3D enzyme plastic nanocomposite structures using a wide range of fabrication techniques, including melt electrowriting, casting, and piston-driven 3D printing. The resulting constructs comprising active phosphotriesterase (arPTE) readily detoxify organophosphates with persistent activity over repeated cycles and for long time periods. Moreover, we show that the protein guest molecules, such as arPTE or sfGFP, increase the compressive Young's modulus of the plastics and that the identity of the biomolecule influences the nanomorphology and mechanical properties of the resulting materials. Overall, we demonstrate that these biologically active nanocomposite plastics are compatible with state-of-the-art 3D fabrication techniques and that the methodology could be readily applied to produce robust and on-demand smart nanomaterial structures.

Fabrication of New Hybrid Scaffolds for in vivo Perivascular Application to Treat Limb Ischemia.

Cell therapies are emerging as a new therapeutic frontier for the treatment of ischemic disease. However, femoral occlusions can be challenging environments for effective therapeutic cell delivery. In this study, cell-engineered hybrid scaffolds are implanted around the occluded femoral artery and the therapeutic benefit through the formation of new collateral arteries is investigated. First, it is reported the fabrication of different hybrid "hard-soft" 3D channel-shaped scaffolds comprising either poly(ε-caprolactone) (PCL) or polylactic-co-glycolic acid (PLGA) and electro-spun of gelatin (GL) nanofibers. Both PCL-GL and PLGA-GL scaffolds show anisotropic characteristics in mechanical tests and PLGA displays a greater rigidity and faster degradability in wet conditions. The resulting constructs are engineered using human adventitial pericytes (APCs) and both exhibit excellent biocompatibility. The 3D environment also induces expressional changes in APCs, conferring a more pronounced proangiogenic secretory profile. Bioprinting of alginate-pluronic gel (AG/PL), containing APCs and endothelial cells, completes the hybrid scaffold providing accurate spatial organization of the delivered cells. The scaffolds implantation around the mice occluded femoral artery shows that bioengineered PLGA hybrid scaffold outperforms the PCL counterpart accelerating limb blood flow recovery through the formation arterioles with diameters >50 µm, demonstrating the therapeutic potential in stimulating reparative angiogenesis.

Flexural resistance of CAD-CAM blocks. Part 3: Polymer-based restorative materials for permanent restorations.

PURPOSE: Concurrently with the growing interest in CAD-CAM systems, several new materials of different chemical nature have become available. As an alternative to ceramics, numerous polymer-based materials have recently been proposed for permanent prosthetic restorations. Aim of this study was to test the CAD-CAM polymer-based materials available on the market, comparing mean flexural strength, Weibull characteristic strength and Weibull modulus. METHODS: Seven types of polymer-based blocks were tested: Lava Ultimate, 3M; Brilliant Crios, Coltene; Cerasmart, GC; Block HC, Shofu; Katana Avencia, Noritake; Grandio Blocs, Voco; Tetric CAD, and Ivoclar-Vivadent. Specimens were cut out from blocks, finished, polished, and tested in a three-point bending test apparatus until failure (n=30). Flexural strength, Weibull characteristic strength, and Weibull modulus were calculated. Flexural strength data were statistically analyzed. ANOVA on Ranks was applied, followed by the Dunn's test for post hoc comparisons (P= 0.05). RESULTS: Flexural strength values (MPa) were measured (mean±standard deviation). Different letters in parentheses label statistically significant differences: Grandio Blocs 266±24(a), Brilliant Crios 259±21(ab); Tetric CAD 254±15(ab); Katana Avencia 241±29(bc); Cerasmart 221±24(cd); Lava Ultimate 196±23(de); Block HC 139±10(e). All the tested materials had flexural strengths greater than 100 MPa, thereby satisfying the requirements of ISO standards for polymer-based materials. For all the tested materials the Weibull characteristic strength was greater than 100 MPa. Weibull modulus ranged between 21.20 (Tetric CAD) and 9.09 (Katana Avencia). CLINICAL SIGNIFICANCE: Even though all the CAD-CAM polymer-based materials marketed in blocks tested in the present study satisfy the requirements of ISO standard for polymer-based materials, their flexural resistance differs significantly. The data presented in the study may be helpful to clinicians for selecting the most appropriate materials for each clinical case.

Influence of Acid Concentration and Etching Time on Composite Cement Adhesion to Lithium-silicate Glass Ceramics.

PURPOSE: To evaluate the influence of hydrofluoric acid (HF) concentration and etching time on the microshear bond strength (µSBS) of RelyX Unicem 2 (3M Oral Care) to VITA Suprinity (Vita Zahnfabrik; zirconia-reinforced lithium-silicate glass ceramic: ZLS) and IPS e.max CAD (Ivoclar Vivadent; lithium-disilicate glass ceramic: LD). MATERIALS AND METHODS: Forty-eight bars each were fabricated of ZLS and LD with Cerec InLab MC-XL. For each material, 8 groups were formed with respect to etching time (20, 40, 60, 120 s) and HF concentration (4.9% and 9.5%). In each group, after etching and silanization, 15 cylindrical specimens of RelyX Unicem 2 were built up for µSBS testing with a special device. Data were analyzed using three-way ANOVA. Failure modes were assessed using an optical stereomicroscope, classified as adhesive, cohesive in resin or ceramic, or mixed. Failure modes were statistically analyzed using Fisher's Exact Test. One bar per group was prepared for SEM observation of the etched surface. RESULTS: ZLS showed significantly higher bond strengths than LD (p < 0.001). Etchant concentration was an influential factor with superior strengths after 4.9% HF etching (p = 0.009). Etching time did not significantly affect adhesion (p = 0.066). The material-etching interaction was statistically significant (p = 0.004). Particularly for ZLS, 4.9% HF etching obtained significantly higher bond strengths than did 9.5%. By using 4.9% HF, adhesion was significantly higher on ZLS than on LD. CONCLUSION: Material and HF concentration influenced the adhesion between RelyX Unicem 2 composite cement and lithium-silicate glass ceramics. Etching time was not an influential factor. The use of 4.9% HF for 20 s proved to be the most effective etching treatment of the intaglio surface.

In Vitro and In Vivo Preclinical Testing of Pericyte-Engineered Grafts for the Correction of Congenital Heart Defects.

Background We have previously reported the possibility of using pericytes from leftovers of palliative surgery of congenital heart disease to engineer clinically certified prosthetic grafts. Methods and Results Here, we assessed the feasibility of using prosthetic conduits engineered with neonatal swine pericytes to reconstruct the pulmonary artery of 9-week-old piglets. Human and swine cardiac pericytes were similar regarding anatomical localization in the heart and antigenic profile following isolation and culture expansion. Like human pericytes, the swine surrogates form clones after single-cell sorting, secrete angiogenic factors, and extracellular matrix proteins and support endothelial cell migration and network formation in vitro. Swine pericytes seeded or unseeded (control) CorMatrix conduits were cultured under static conditions for 5 days, then they were shaped into conduits and incubated in a flow bioreactor for 1 or 2 weeks. Immunohistological studies showed the viability and integration of pericytes in the outer layer of the conduit. Mechanical tests documented a reduction in stiffness and an increase in strain at maximum load in seeded conduits in comparison with unseeded conduits. Control and pericyte-engineered conduits were then used to replace the left pulmonary artery of piglets. After 4 months, anatomical and functional integration of the grafts was confirmed using Doppler echography, cardiac magnetic resonance imaging, and histology. Conclusions These findings demonstrate the feasibility of using neonatal cardiac pericytes for reconstruction of small-size branch pulmonary arteries in a large animal model.

Zirconia Translucency and Cement Systems as Factors Influencing the Zirconia-Titanium and Zirconia-Zirconia Shear Bond Strength.

PURPOSE: To evaluate the bonding ability of self-adhesive and dual-cure resin cement systems applied to zirconia materials of different translucency in combination with each other and with titanium. MATERIALS AND METHODS: Computer-aided design/computer-assisted manufacture (CAD/CAM) disks of three different zirconia materials (Zenostar MO 0 [medium opacity], MT 0 [medium translucency], and T 1 [translucent], Wieland Dental) were milled and hard sintered in order to obtain cylindrical samples of either 12 mm high × 12 mm diameter (used as a substrate) or 12 mm high × 8 mm diameter (to be bonded to the substrate). CAD/CAM disks of titanium (Starbond Ti4 Disc, S&S Scheftner) were milled in order to obtain a cylinder shape (12 mm high × 8 mm diameter to be bonded to the zirconia substrate). The bonding surfaces were sandblasted with alumina particles (70 µm/2 ATM). Two resin cement systems were tested in this study: a self-adhesive resin cement (SpeedCEM Plus, Ivoclar Vivadent) and a dual-cure resin cement (Variolink Esthetic DC, Ivoclar Vivadent) in combination with a primer (Monobond Plus, Ivoclar Vivadent). The smaller cylinders were bonded to the substrate following the manufacturer's instructions. After 24 hours at 37°C storage, each specimen was subjected to shear bond strength testing in a universal testing machine. The substrate was fixed to the machine, and shear force was applied to the small cylinder close to the bonding interface. Shear bond strength data were statistically analyzed by a three-way analysis of variance (ANOVA) followed by the Tukey test for post hoc comparison (P < .05). RESULTS: No significant differences were found between substrates in the shear bond strength test (P = .078). Variolink Esthetic DC showed a statistically significantly higher bond strength compared with the SpeedCEM Plus (P = .001) with the exception of Zenostar MT substrate (P = .014). CONCLUSION: The resin cement systems showed high shear bond strength values when zirconia was resin bonded to either titanium or the corresponding zirconia substrate. Different zirconia translucencies did not affect the bond strength. The use of MDP primer and a dual-cure cement showed statistically significantly higher strength compared with the self-adhesive resin cement system with the only exception being the bonding of the translucent zirconia Zenostar MT.

The Effect of Matrix Stiffness of Biomimetic Gelatin Nanofibrous Scaffolds on Human Cardiac Pericyte Behavior.

Congenital heart disease (CHD) is the most common and deadly congenital anomaly, accounting for up to 7.5% of all infant deaths. Survival in children born with CHD has improved dramatically over the past several decades (this positive trend being counterbalanced by the fact that more patients develop heart failure). Seminal data indicate an alteration of the extracellular matrix occurs with time in these hearts due to diffuse and abundant interstitial fibrosis. This results in an escalation in the stiffness of the local myocardial microenvironment. However, the influence of matrix stiffness in regulating the function of resident human stromal cells has not been reported. The objective of this study was to determine the impact of scaffold stiffness on the antigenic and functional profile of cardiac pericytes (CPs) isolated from patients with CHD. To this end, we have first manufactured gelatin nanofibrous scaffolds with varying degrees of stiffness using an in situ cross-linking electrospinning technique in a pure water solvent system. We assessed Young's modulus and performed a comprehensive physicochemical characterization of the scaffolds employing scanning electron microscopy and Fourier transform infrared spectroscopy. We next evaluated the changes induced by a different scaffold stiffness on CP morphology, antigenic profile, viability, proliferation, angiocrine activity, and induced differentiation. Results indicate that soft matrixes with a fiber diameter of ∼400 nm increase CP proliferation, secretion of angiopoietin 2, and F-actin stress fiber formation, without affecting the antigenic profile, viability, or differentiation. These data indicate for the first time that human CPs can be functionally influenced by slight changes in matrix stiffness. The study elucidates the importance of mechanical/morphological cues in modulating the behavior of stromal cells isolated from patients with CHD.

Effects of scanning technique on in vitro performance of CAD/CAM-fabricated fiber posts.

This study assessed push-out strength, cement layer thickness, and interfacial nanoleakage of luted fiber posts fabricated with computer-aided design/computer-assisted manufacture (CAD/CAM) technology after use of 1 of 3 scanning techniques, namely, direct scanning of the post space (DS), scanning of a polyether impression of the post space (IS), and scanning of a plaster model of the post space (MS). Thirty premolars were randomly assigned to three groups corresponding to the scanning technique. Posts were computer-designed and milled from experimental fiber-reinforced composite blocks. The mean (±SD) values for push-out strength and cement thickness were 17.1 ± 7.7 MPa and 162 ± 24 µm, respectively, for DS, 10.7 ± 4.6 MPa and 187 ± 50 µm for IS, and 12.0 ± 7.2 MPa and 258 ± 78 µm for MS specimens. Median (interquartile range) interfacial nanoleakage scores were 3 (2-4) for DS, 2.5 (2-4) for IS, and 3 (2-4) for MS. Post retention was better for fiber posts fabricated by DS technique than for those fabricated by IS and MS. Cement thickness did not differ between DS and IS specimens, but the cement layer was significantly thicker in the MS group than in the other two groups. Scanning technique did not affect sealing ability, as the three groups had comparable nanoleakage values.