Impact of In-Process Crystallinity of Biodegradable Scaffolds Fabricated by Material Extrusion on the Micro- and Nanosurface Topography, Viability, Proliferation, and Differentiation of Human Mesenchymal Stromal Cells.

Due to affordability, and the ability to parametrically control the vital processing parameters, material extrusion is a widely accepted technology in tissue engineering. Material extrusion offers sufficient control over pore size, geometry, and spatial distribution, and can also yield different levels of in-process crystallinity in the resulting matrix. In this study, an empirical model based on four process parameters-extruder temperature, extrusion speed, layer thickness, and build plate temperature-was used to control the level of in-process crystallinity of polylactic acid (PLA) scaffolds. Two sets of scaffolds were fabricated, with low- and high-crystallinity content, and subsequently seeded with human mesenchymal stromal cells (hMSC). The biochemical activity of hMSC cells was tested by examining the DNA content, lactate dehydrogenase (LDH) activity, and alkaline phosphatase (ALP) tests. The results of this 21-day in vitro experiment showed that high level crystallinity scaffolds performed significantly better in terms of cell response. Follow-up tests revealed that the two types of scaffolds were equivalent in terms of hydrophobicity, and module of elasticity. However, detailed examination of their micro- and nanosurface topographic features revealed that the higher crystallinity scaffolds featured pronounced nonuniformity and a larger number of summits per sampling area, which was the main contributor to a significantly better cell response.

Finite Element Simplifications and Simulation Reliability in Single Point Incremental Forming.

Single point incremental forming (SPIF) is one of the most promising technologies for the manufacturing of sheet metal prototypes and parts in small quantities. Similar to other forming processes, the design of the SPIF process is a demanding task. Nowadays, the design process is usually performed using numerical simulations and virtual models. The modelling of the SPIF process faces several challenges, including extremely long computational times caused by long tool paths and the complexity of the problem. Path determination is also a demanding task. This paper presents a finite element (FE) analysis of an incrementally formed truncated pyramid compared to experimental validation. Focus was placed on a possible simplification of the FE process modelling and its impact on the reliability of the results obtained, especially on the geometric accuracy of the part and bottom pillowing effect. The FE modelling of SPIF process was performed with the software ABAQUS, while the experiment was performed on a conventional milling machine. Low-carbon steel DC04 was used. The results confirm that by implementing mass scaling and/or time scaling, the required calculation time can be significantly reduced without substantially affecting the pillowing accuracy. An innovative artificial neural network (ANN) approach was selected to find the optimal values of mesh size and mass scaling in term of minimal bottom pillowing error. However, care should be taken when increasing the element size, as it has a significant impact on the pillow effect at the bottom of the formed part. In the range of selected mass scaling and element size, the smallest geometrical error regarding the experimental part was obtained by mass scaling of 19.01 and tool velocity of 16.49 m/s at the mesh size of 1 × 1 mm. The obtained results enable significant reduction of the computational time and can be applied in the future for other incrementally formed shapes as well.

Layer-by-layer bioassembly of poly(lactic) acid membranes loaded with coculture of HBMSCs and EPCs improves vascularization in vivo.

Layer-by-layer (LBL) BioAssembly method was developed to enhance the control of cell distribution within 3D scaffolds for tissue engineering applications. The objective of this study was to evaluate in vivo the development of blood vessels within LBL bioassembled membranes seeded with human primary cells, and to compare it to cellularized massive scaffolds. Poly(lactic) acid (PLA) membranes fabricated by fused deposition modeling were seeded with monocultures of human bone marrow stromal cells or with cocultures of these cells and endothelial progenitor cells. Then, four cellularized membranes were assembled in LBL constructs. Early osteoblastic and endothelial cell differentiation markers, alkaline phosphatase, and von Willebrand's factor, were expressed in all layers of assemblies in homogenous manner. The same kind of LBL assemblies as well as cellularized massive scaffolds was implanted subcutaneously in mice. Human cells were observed in all scaffolds seeded with cells, but not in the inner parts of massive scaffolds. There were significantly more blood vessels observed in LBL bioassemblies seeded with cocultures compared to all other samples. LBL bioassembly of PLA membranes seeded with a coculture of human cells is an efficient method to obtain homogenous cell distribution and blood vessel formation within the entire volume of a 3D composite scaffold.

Influence of the restorative procedure factors on stress values in premolar with MOD cavity: a finite element study.

In order to investigate the influence of cusp reduction, cavity isthmus width, and restorative material on stress values in premolar with mesio-occlusal-distal (MOD) cavity, numerical simulations were done on three-dimensional (3D) models of a maxillary second premolar designed using computerized tomography (CT) scan images. The use of four restorative materials (direct resin composite, direct resin composite with resin-modified glass-ionomer cement as the base, indirect resin composite, ceramic), three cavity preparation designs (without cusp coverage, 2-mm palatal cusp coverage, 2-mm palatal and buccal cusp coverage), and two cavity isthmus widths (1/2 and 2/3 intercuspal width) were simulated. After applying a static load of 200 N on the occlusal surface of the tooth, von Mises stresses in the enamel, dentin, and restoration were calculated using finite element analysis (FEA). Stress values in the enamel were primarily influenced by cavity preparation design, while restorative material showed higher contribution in dentin. The lowest stress values were obtained in models with cusp coverage and indirect restorations. Cavity isthmus width had minimal influence on stress values in tooth structures. None of the investigated factors determined stress values in the restoration. In conclusion, the use of ceramic restoration covering both palatal and buccal cusp provided the most favourable stress distribution of premolars with MOD cavity. Graphical abstract ᅟ.

Influence of restorative procedures on endodontically treated premolars: Finite element analysis of a CT-scan based three-dimensional model.

An endodontically treated tooth with mesial-occlusal-distal (MOD) cavity is often restored with composite resin. Palatal and buccal cusp reduction (MODP, MODPB), and/or fiber-reinforced composite posts (P), are used in an attempt to improve the longevity of the restoration. The aim of this study was to determine the effects of these procedures on von Mises stress values and distribution in dental tissues and restorative materials using finite element analysis. Based on CT scans of an extracted second upper premolar, six 3D endodontically treated tooth models (MOD, MODP, MODPB, MOD+P, MODP+P, MODPB+P) were created. Each model was subjected to a summary force of 150 N on the occlusal surface simulating the normal biting pattern and maximal von Mises stresses were calculated. MODP seems to reduce von Mises stress values in dental tissues and P seems to transfer some of the stresses from dental tissues to the composite filling.

Layer-by-layer bioassembly of cellularized polylactic acid porous membranes for bone tissue engineering.

The conventional tissue engineering is based on seeding of macroporous scaffold on its surface ("top-down" approach). The main limitation is poor cell viability in the middle of the scaffold due to poor diffusion of oxygen and nutrients and insufficient vascularization. Layer-by-Layer (LBL) bioassembly is based on "bottom-up" approach, which considers assembly of small cellularized blocks. The aim of this work was to evaluate proliferation and differentiation of human bone marrow stromal cells (HBMSCs) and endothelial progenitor cells (EPCs) in two and three dimensions (2D, 3D) using a LBL assembly of polylactic acid (PLA) scaffolds fabricated by 3D printing. 2D experiments have shown maintain of cell viability on PLA, especially when a co-cuture system was used, as well as adequate morphology of seeded cells. Early osteoblastic and endothelial differentiations were observed and cell proliferation was increased after 7 days of culture. In 3D, cell migration was observed between layers of LBL constructs, as well as an osteoblastic differentiation. These results indicate that LBL assembly of PLA layers could be suitable for BTE, in order to promote homogenous cell distribution inside the scaffold and gene expression specific to the cells implanted in the case of co-culture system.

Poly(methyl-methacrylate) nanocomposites with low silica addition.

STATEMENT OF PROBLEM: Poly(methyl-methacrylate) (PMMA) represents the most popular current denture material. However, its major drawbacks are insufficient ductility and strength. PURPOSE: The purpose of this study was to improve the mechanical properties of PMMA in denture base application by adding small quantities of nanosilica. MATERIAL AND METHOD: Silica nanoparticles were added to the liquid component of the tested materials. The standard heat polymerizing procedure was followed to obtain 6 PMMA--silicon dioxide (/SiO2) concentrations (0.023%, 0.046%, 0.091%, 0.23%, 0.46%, and 0.91% by volume). Microhardness and fracture toughness of each set of specimens was compared with the unmodified specimens. Furthermore, differential scanning calorimetry and scanning electron microscopy analyses were conducted, and the results obtained were correlated with the results of mechanical properties. RESULTS: It was found that the maximum microhardness and fracture toughness values of the materials tested were obtained for the lowest nanosilica content. A nanosilica content of 0.023% resulted in an almost unchanged glass transition temperature (Tg), whereas the maximum amount of nanosilica induced a considerable increase in Tg. A higher Tg indicated the possible existence of a thicker interfacial layer caused by the chain immobility due to the presence of the particles. However, scanning electron microscopy results demonstrated extensive agglomeration at 0.91% nanosilica, which may have prevented the formation of a homogenous reinforced field. At a nanosilica content of 0.023%, no agglomeration was observed, which probably influenced a more homogenous distribution of nanoparticles as well as uniform reinforcing fields. CONCLUSIONS: Low nanoparticle content yields superior mechanical properties along with the lower cost of nanocomposite synthesis.

Influence of cavity design preparation on stress values in maxillary premolar: a finite element analysis.

AIM: To analyze the influence of cavity design preparation on stress values in three-dimensional (3D) solid model of maxillary premolar restored with resin composite. METHODS: 3D solid model of maxillary second premolar was designed using computed-tomography (CT) data. Based on a factorial experiment, 9 different mesio-occlusal-distal (MOD) cavity designs were simulated, with three cavity wall thicknesses (1.5 mm, 2.25 mm, 3.0 mm), and three cusp reduction procedures (without cusp reduction, 2.0 mm palatal cusp reduction, 2.0 mm palatal and buccal cusp reduction). All MOD cavities were simulated with direct resin composite restoration (Gradia Direct Posterior, GC, Japan). Finite element analysis (FEA) was used to calculate von Mises stress values. RESULTS: The von Mises stresses in enamel, dentin, and resin composite were 79.3-233.6 MPa, 26.0-32.9 MPa, and 180.2-252.2 MPa, respectively. Considering the influence of cavity design parameters, cuspal reduction (92.97%) and cavity wall thickness (3.06%) significantly (P<0.05) determined the magnitude of stress values in enamel. The influence of cavity design parameters on stress values in dentin and resin composite was not significant. When stresses for enamel, dentine, and resin composite were considered all together, palatal cusp coverage was revealed as an optimal option. Cavity wall thickness did not show a significant effect on stress values. CONCLUSION: Based on numerical simulations, a palatal cusp reduction could be suggested for revealing lower stress values in dental tissues and restorative material. This type of cavity design should contribute to better biomechanical behavior of tooth-restoration complex, consequently providing the long-lasting clinical results.