Micro-CT imaging and finite element models reveal how sintering temperature affects the microstructure and strength of bioactive glass-derived scaffolds.

This study focuses on the finite element simulation and micromechanical characterization of bioactive glass-ceramic scaffolds using Computed micro Tomography ([Formula: see text]CT) imaging. The main purpose of this work is to quantify the effect of sintering temperature on the morphometry and mechanical performance of the scaffolds. In particular, the scaffolds were produced using a novel bioactive glass material (47.5B) through foam replication, applying six different sintering temperatures. Through [Formula: see text]CT imaging, detailed three-dimensional images of the scaffold's internal structure are obtained, enabling the extraction of important geometric features and how these features change with sintering temperature. A finite element model is then developed based on the [Formula: see text]CT images to simulate the fracture process under uniaxial compression loading. The model incorporates scaffold heterogeneity and material properties-also depending on sintering temperature-to capture the mechanical response, including crack initiation, propagation, and failure. Scaffolds sintered at temperatures equal to or higher than 700 [Formula: see text]C exhibit two-scale porosity, with micro and macro pores. Finite element analyses revealed that the dual porosity significantly affects fracture mechanisms, as micro-pores attract cracks and weaken strength. Interestingly, scaffolds sintered at high temperatures, the overall strength of which is higher due to greater intrinsic strength, showed lower normalized strength compared to low-temperature scaffolds. By using a combined strategy of finite element simulation and [Formula: see text]CT-based characterization, bioactive glass-ceramic scaffolds can be optimized for bone tissue engineering applications by learning more about their micromechanical characteristics and fracture response.

Computational models for the simulation of the elastic and fracture properties of highly porous 3D-printed hydroxyapatite scaffolds.

Bone scaffolding is a promising approach for the treatment of critical-size bone defects. Hydroxyapatite can be used to produce highly porous scaffolds as it mimics the mineralized part of bone tissue, but its intrinsic brittleness limits its usage. Among 3D printing techniques, vat photopolymerization allows for the best printing resolution for ceramic materials. In this study, we implemented a Computed micro-Tomography based Finite Element Model of a hydroxyapatite porous scaffold fabricated by vat photopolymerization. We used the model in order to predict the elastic and fracture properties of the scaffold. From the stress-strain diagram of a simulated compression test, we computed the stiffness and the strength of the scaffolds. We found that three morphometric features substantially affect the crack pattern. In particular, the crack propagation is not only dependent on the trabecular thickness but also depends on the slenderness and orientation of the trabeculae with respect to the load. The results found in this study can be used for the design of ceramic scaffolds with heterogeneous pore distribution in order to tailor and predict the compressive strength.

Mechanical Properties of Robocast Glass Scaffolds Assessed through Micro-CT-Based Finite Element Models.

In this study, the mechanical properties of two classes of robocast glass scaffolds are obtained through Computed micro-Tomography (micro-CT) based Finite Element Modeling (FEM) with the specific purpose to explicitly account for the geometrical defects introduced during manufacturing. Both classes demonstrate a fiber distribution along two perpendicular directions on parallel layers with a 90∘ tilting between two adjacent layers. The crack pattern identified upon compression loading is consistent with that found in experimental studies available in literature. The finite element models have demonstrated that the effect of imperfections on elastic and strength properties may be substantial, depending on the specific type of defect identified in the scaffolds. In particular, micro-porosity, fiber length interruption and fiber detaching were found as key factors. The micro-pores act as stress concentrators promoting fracture initiation and propagation, while fiber detachment reduces the scaffold properties substantially along the direction perpendicular to the fiber plane.

Mineralization in a Critical Size Bone-Gap in Sheep Tibia Improved by a Chitosan-Calcium Phosphate-Based Composite as Compared to Predicate Device.

Deacetylated chitin derivatives have been widely studied for tissue engineering purposes. This study aimed to compare the efficacy of an injectable product containing a 50% deacetylated chitin derivative (BoneReg-Inject™) and an existing product (chronOS Inject®) serving as a predicate device. A sheep model with a critical size drill hole in the tibial plateau was used. Holes of 8 mm diameter and 30 mm length were drilled bilaterally into the proximal area of the tibia and BoneReg-Inject™ or chronOS Inject® were injected into the right leg holes. Comparison of resorption and bone formation in vivo was made by X-ray micro-CT and histological evaluation after a live phase of 12 weeks. Long-term effects of BoneReg-Inject™ were studied using a 13-month live period. Significant differences were observed in (1) amount of new bone within implant (p < 0.001), higher in BoneReg-InjectTM, (2) signs of cartilage tissue (p = 0.003), more pronounced in BoneReg-InjectTM, and (3) signs of fibrous tissue (p < 0.001), less pronounced in BoneReg-InjectTM. Mineral content at 13 months postoperative was significantly higher than at 12 weeks (p < 0.001 and p < 0.05, for implant core and rim, respectively). The data demonstrate the potential of deacetylated chitin derivatives to stimulate bone formation.

Polyphenols from Grape Pomace: Functionalization of Chitosan-Coated Hydroxyapatite for Modulated Swelling and Release of Polyphenols.

Chitosan is known for its specific antibacterial mechanism and biodegradability, while polyphenols are known for their antioxidant and anti-inflammatory properties: coupling these properties on a surface for bone contact, such as hydroxyapatite, is of great interest. The system developed here allows the combination of hydroxyapatite, chitosan, and polyphenol properties in the same multifunctional biomaterial in order to modulate the host response after implantation. Crosslinked chitosan is used in this research to create a stable coating on hydroxyapatite, and then it is functionalized for a smart release of the polyphenols. The release is higher in inflammatory conditions and lower in physiological conditions. The properties of the coated and functionalized samples are characterized on the as-prepared samples and after the samples are immersed (for 24 h) in solutions, which simulate the inflammatory and physiological conditions. Characterization is performed in order to confirm the presence of polyphenols grafted within the chitosan coating, the stability of grafting as a function of pH, the morphology of the coating and distribution of polyphenols on the surface, and the redox reactivity and radical scavenging activity of the functionalized coating. All the results are in line with previous results, which show a successful coating with chitosan and functionalization with polyphenols. Moreover, the polyphenols have a different release kinetics that is faster in a simulated inflammatory environment compared to that in the physiological environment. Even after the release tests, a fraction of polyphenols are still bound on the surface, maintaining the antioxidant and radical scavenging activity for a longer time. An electrostatic bond occurs between the negative-charged polar groups of polyphenols (carboxyls and/or phenols) and the positive amide groups of the chitosan coating, and the substitution of the crosslinker by the polyphenols occurs during the functionalization process.

Comprehensive assessment of bioactive glass and glass-ceramic scaffold permeability: experimental measurements by pressure wave drop, modelling and computed tomography-based analysis.

Proper microstructural and transport properties are fundamental requirements for a suitable scaffold design and realization in tissue engineering applications. Scaffold microstructure (i.e. pore size, shape and distribution) and transport properties (i.e. intrinsic permeability), are commonly recognized as the key parameters related to the biological performance, such as cell attachment, penetration depth and tissue vascularization. While pore characteristics are relatively easy to asses, accurate and reliable evaluation of permeability still remains a challenge. In the present study, the microstructural properties of foam-replicated bioactive glass-derived scaffolds (basic composition 47.5SiO2-2.5P2O5-20CaO-10MgO-10Na2O-10K2O mol.%) were determined as function of the sintering temperature within the range 600-850°C, identified on the basis of thermal analyses that were previously performed on the material. Scaffolds with total porosity between 55 and 84 vol.% and trabecular-like architecture were obtained, with pore morphological features varying according to the sintering temperature. Mathematical modelling, supported by micro-computed tomography (µ-CT) imaging, was implemented to selectively investigate the effect of different pore features on intrinsic permeability, which was determined by laminar airflow alternating pressure wave drop measurements and found to be within 0.051-2.811·10-10 m2. The calculated effective porosity of the scaffolds was in the range of 46 to 66 vol.%, while the average pore diameter assessed by µ-CT varied between 220 and 780 µm, where the values in the lower range were observed for higher sintering temperatures (750-850°C). Experimental results were critically discussed by means of a robust statistical analysis. Finally, the complete microstructural characterization of the scaffolds was achieved by applying the general constitutive equation based on Forchheimer's theory.

Bone remodeling effect of a chitosan and calcium phosphate-based composite.

Chitosan is a biocompatible polymer that has been widely studied for tissue engineering purposes. The aim of this research was to assess bone regenerative properties of an injectable chitosan and calcium phosphate-based composite and identify optimal degree of deacetylation (%DDA) of the chitosan polymer. Drill holes were generated on the left side of a mandible in Sprague-Dawley rats, and the hole was either left empty or filled with the implant. The animals were sacrificed at several time points after surgery (7-22 days) and bone was investigated using micro-CT and histology. No significant new bone formation was observed in the implants themselves at any time points. However, substantial new bone formation was observed in the rat mandible further away from the drill hole. Morphological changes indicating bone formation were found in specimens explanted on Day 7 in animals that received implant. Similar bone formation pattern was seen in control animals with an empty drill hole at later time points but not to the same extent. A second experiment was performed to examine if the %DDA of the chitosan polymer influenced the bone remodeling response. The results suggest that chitosan polymers with %DDA between 50 and 70% enhance the natural bone remodeling mechanism.

Robocasting of SiO2-Based Bioactive Glass Scaffolds with Porosity Gradient for Bone Regeneration and Potential Load-Bearing Applications.

: Additive manufacturing of bioactive glasses has recently attracted high interest in the field of regenerative medicine as a versatile class of fabrication methods to process bone substitute materials. In this study, melt-derived glass particles from the SiO2-P2O5-CaO-MgO-Na2O-K2O system were used to fabricate bioactive scaffolds with graded porosity by robocasting. A printable ink made of glass powder and Pluronic F-127 (binder) was extruded into a grid-like three-dimensional structure with bimodal porosity, i.e., the inner part of the scaffold had macropores with smaller size compared to the periphery. The crystallization behavior of the glass powder was studied by hot-stage microscopy, differential thermal analysis, and X-ray diffraction; the scaffolds were sintered at a temperature below the onset of crystallization so that amorphous structures could be obtained. Scaffold architecture was investigated by scanning electron microscopy and microtomographic analysis that allowed quantifying the microstructural parameters. In vitro tests in Kokubo's simulated body fluid (SBF) confirmed the apatite-forming ability (i.e., bioactivity) of the scaffolds. The compressive strength was found to slightly decrease during immersion in SBF up to 4 weeks but still remained comparable to that of human cancellous bone. The pH and concentration of released ions in SBF were also measured at each time point. Taken together, these results (favorable porosity, mechanical strength, and in vitro bioactivity) show great promise for the potential application of these robocast scaffolds in bone defect repair.

Robocasting of Bioactive SiO2-P2O5-CaO-MgO-Na2O-K2O Glass Scaffolds.

Bioactive silicate glass scaffolds were fabricated by a robocasting process in which all the movements of the printing head were programmed by compiling a script (text file). A printable ink made of glass powder and Pluronic F-127, acting as a binder, was extruded to obtain macroporous scaffolds with a grid-like three-dimensional structure. The scaffold architecture was investigated by scanning electron microscopy and microtomographic analysis, which allowed quantifying the microstructural parameters (pore size 150-180 µm and strut diameter 300 µm). In vitro tests in simulated body fluid (SBF) confirmed the apatite-forming ability (i.e., bioactivity) of the scaffolds. The compressive strength (around 10 MPa for as-produced scaffolds) progressively decreased during immersion in SBF (3.3 MPa after 4 weeks) but remains acceptable for bone repair applications. Taken together, these results (adequate porosity and mechanical strength as well as bioactivity) support the potential suitability of the prepared scaffolds for bone substitution.

Solution casting of chitosan membranes for in vitro evaluation of bioactivity.

BACKGROUND: Considerable research is focusing on the surface modification of titanium implants for the treatment of orthopaedic tissue injuries to increase the success of orthopaedic fixations. Chitosan is one of the natural materials under investigation based on several favourable properties. Numerous techniques have been described for the preparation of chitosan membranes, including solution casting methods for the investigation of bioactivity before applying coatings onto potential titanium implants. Solution casting enables the easy in-house evaluation of chitosan membranes and allows for the selection of promising chitosan materials. RESULTS: We present a method for the standardized and easily applied preparation of chitosan membranes by solution casting. This protocol is suitable for chitosan materials spanning a wide degree of deacetylation, being derived from different chitin sources and chitosan derivatives with novel properties. We detail the preparation and quality control methods in order to prepare membranes with favourable bioactivity, sustaining cell attachment and proliferation for extended culture periods. CONCLUSIONS: The possibilities associated with the use of chitosan in tissue engineering applications are far from being exhausted and numerous challenges remain prior to successful translation into the clinics. Based on our experience, we have developed simple in-house methods for quality control of homogeneous membrane casting and early prediction of successful experimental outcome.

In vitro bioactivity of different degree of deacetylation chitosan, a potential coating material for titanium implants.

Clinical treatment of orthopaedic tissue injuries often involves the use of titanium and titanium alloys with considerable research focusing on the surface modification of these materials. Chitosan, the partly deacetylated form of chitin, is one of the materials under investigation as surface coating for orthopaedic implants in order to improve osteo-integration and cellular attachment. In this study, we determined the effects of the degree of deacetylation (DD) of chitosan membranes on attachment, proliferation and osteogenic differentiation of MC3T3-E1 mouse preosteoblasts. Chitosan membranes were coated with fibronectin to promote biocompatibility and cellular attachment. Membranes were characterized in terms of wettability and surface topography using water contact angle measurements and atomic force microscopy. The results in this study indicate that the surface roughness and fibronectin adsorption increase with increased DD. A higher DD also facilitates attachment and proliferation of cells, but no induction of spontaneous osteogenic differentiation was observed. Lower DD chitosan membranes were successfully prepared to sustain attachment and were modified by crosslinking with glutaraldehyde to promote long-term studies. The chitosan membranes used in this study are suitable as a potential coating for titanium implants.

Synthesis and Crystal Structure of Zr(2)Te. Distinctions in Bonding to Isotypic Sc(2)Te and the Relationship to the Structures of Congeneric Hf(2)Te and Zr(2)Se.

Zr(2)Te is accessible by high-temperature synthesis. The structure of the zirconium-rich telluride was determined by means of powder X-ray diffraction to be orthorhombic, Pnma (No. 62), Z = 12, Pearson symbol oP36, a = 1995.0(2) pm, b = 382.36(2) pm, c = 1065.63(9) pm. Pairwise interpenetrating columns of trans-face-shared, centered Zr(9) cuboids, reminiscent of the bcc high-temperature form of zirconium can be recognized as the topologically characteristic structural feature. Tellurium atoms capping the remaining square faces complete the motif of a [Zr(8)Te(4)] double string running parallel [010]. The tellurium atoms are 7-, 8- and 9-fold coordinated by zirconium. The coordination figures represent mono-, bi- and tricapped distorted trigonal prisms, with zirconium atoms capping the square faces of the prisms. Extended Hückel calculations revealed distinctions in bonding in Zr(2)Te and the isotypic Sc(2)Te. According to Mulliken overlap populations, the heteronuclear interactions are similar in both tellurides. However, the lower valence electron concentration available for M-M bonding in Sc(2)Te is reflected in a considerable restriction of the attractive homonuclear interactions to one-dimensional metal cores, whereas in Zr(2)Te M-M bonding regions extend in space. The structure of Zr(2)Te is contrasted with two other types of bcc fragment structures adopted by the congeneric Hf(2)Te and Zr(2)Se. We show that the structural diversity observed for various dimetal chalcogenides is controlled by an intimate interplay of electronic and geometric factors.