Osteogenic Properties of 3D-Printed Silica-Carbon-Calcite Composite Scaffolds: Novel Approach for Personalized Bone Tissue Regeneration.

Carbon enriched bioceramic (C-Bio) scaffolds have recently shown exceptional results in terms of their biological and mechanical properties. The present study aims at assessing the ability of the C-Bio scaffolds to affect the commitment of canine adipose-derived mesenchymal stem cells (cAD-MSCs) and investigating the influence of carbon on cell proliferation and osteogenic differentiation of cAD-MSCs in vitro. The commitment of cAD-MSCs to an osteoblastic phenotype has been evaluated by expression of several osteogenic markers using real-time PCR. Biocompatibility analyses through 3-(4,5-dimethyl- thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), lactate dehydrogenase (LDH) activity, hemolysis assay, and Ames test demonstrated excellent biocompatibility of both materials. A significant increase in the extracellular alkaline phosphatase (ALP) activity and expression of runt-related transcription factor (RUNX), ALP, osterix (OSX), and receptor activator of nuclear factor kappa-Β ligand (RANKL) genes was observed in C-Bio scaffolds compared to those without carbon (Bio). Scanning electron microscopy (SEM) demonstrated excellent cell attachment on both material surfaces; however, the cellular layer on C-Bio fibers exhibited an apparent secretome activity. Based on our findings, graphene can improve cell adhesion, growth, and osteogenic differentiation of cAD-MSCs in vitro. This study proposed carbon as an additive for a novel three-dimensional (3D)-printable biocompatible scaffold which could become the key structural material for bone tissue reconstruction.

Waste Glass Upcycling Supported by Alkali Activation: An Overview.

Alkali-activated materials are gaining much interest due to their outstanding performance, including their great resistance to chemical corrosion, good thermal characteristics, and ability to valorise industrial waste materials. Reusing waste glasses in creating alkali-activated materials appears to be a viable option for more effective solid waste utilisation and lower-cost products. However, very little research has been conducted on the suitability of waste glass as a prime precursor for alkali activation. This study examines the reuse of seven different types of waste glasses in the creation of geopolymeric and cementitious concretes as sustainable building materials, focusing in particular on how using waste glasses as the raw material in alkali-activated materials affects the durability, microstructures, hydration products, and fresh and hardened properties in comparison with using traditional raw materials. The impacts of several vital parameters, including the employment of a chemical activator, gel formation, post-fabrication curing procedures, and the distribution of source materials, are carefully considered. This review will offer insight into an in-depth understanding of the manufacturing and performance in promising applications of alkali-activated waste glass in light of future uses. The current study aims to provide a contemporary review of the chemical and structural properties of glasses and the state of research on the utilisation of waste glasses in the creation of alkali-activated materials.

Upcycling waste derived glass into high-performance photocatalytic scaffolds by alkali activation and direct ink writing.

Novel and eco-friendly solutions are extensively needed for wastewater treatment. This work capitalizes on the combination of waste vitrification and additive manufacturing to produce an efficient photocatalyst for the specific purpose. Fine powders of waste-derived glass, containing Fe3O4 inclusions, by simple suspension (for a solid loading of 65 wt %) in alkaline solution (5 M NaOH), were transformed into pastes for direct ink writing. 3D-printed reticulated scaffolds were stabilized by the progressive hardening of a zeolite-like gel, formed by glass/solution interaction, at nearly room temperature. The printed scaffolds were successfully tested for the removal of methylene blue, realized by combining the high sorption capacity of the gel with the catalytic activity of magnetite inclusions, under UV light. A complete degradation of methylene blue is achieved by 90 min exposure, comparing favorably with other reported photocatalytic materials, requiring from 60 to 360 min. The photocatalytic activity was tested for several cycles, with no significant degradation. In other words, a waste-derived material can be reused for multiple times, to remediate wastewaters, with evident benefits on waste minimization.

Modeling Materials Coextrusion in Polymers Additive Manufacturing.

Material extrusion additive manufacturing enables us to combine more materials in the same nozzle during the deposition process. This technology, called material coextrusion, generates an expanded range of material properties, which can gradually change in the design domain, ensuring blending or higher bonding/interlocking among the different materials. To exploit the opportunities offered by these technologies, it is necessary to know the behavior of the combined materials according to the materials fractions. In this work, two compatible pairs of materials, namely Polylactic Acid (PLA)-Thermoplastic Polyurethane (TPU) and Acrylonitrile Styrene Acrylate (ASA)-TPU, were investigated by changing the material fractions in the coextrusion process. An original model describing the distribution of the materials is proposed. Based on this, the mechanical properties were investigated by analytical and numerical approaches. The analytical model was developed on the simplified assumption that the coextruded materials are a set of rods, whereas the more realistic numerical model is based on homogenization theory, adopting the finite element analysis of a representative volume element. To verify the deposition model, a specific experimental test was developed, and the modeled material deposition was superimposed and qualitatively compared with the actual microscope images regarding the different deposition directions and material fractions. The analytical and numerical models show similar trends, and it can be assumed that the finite element model has a more realistic behavior due to the higher accuracy of the model description. The elastic moduli obtained by the models was verified in experimental tensile tests. The tensile tests show Young's moduli of 3425 MPa for PLA, 1812 MPa for ASA, and 162 MPa for TPU. At the intermediate material fraction, the Young's modulus shows an almost linear trend between PLA and TPU and between ASA and TPU. The ultimate tensile strength values are 63.9 MPa for PLA, 35.7 MPa for ASA, and 63.5 MPa for TPU, whereas at the intermediate material fraction, they assume lower values. In this initial work, the results show a good agreement between models and experiments, providing useful tools for designers and contributing to a new branch in additive manufacturing research.

Proteolytically Resistant Bioactive Peptide-Grafted Sr/Mg-Doped Hardystonite Foams: Comparison of Two Covalent Functionalization Strategies.

Hardystonite-based (HT) bioceramic foams were easily obtained via thermal treatment of silicone resins and reactive oxide fillers in air. By using a commercial silicone, incorporating strontium oxide and magnesium oxide precursors (as well as CaO and ZnO), and treating it at 1100 °C, a complex solid solution (Ca1.4Sr0.6Zn0.85Mg0.15Si2O7) that has superior biocompatibility and bioactivity properties compared to pure hardystonite (Ca2ZnSi2O7) can be obtained. Proteolytic-resistant adhesive peptide mapped on vitronectin (D2HVP), was selectively grafted to Sr/Mg-doped HT foams using two different strategies. Unfortunately, the first method (via protected peptide) was unsuitable for acid-sensitive materials such as Sr/Mg-doped HT, resulting in the release of cytotoxic levels of Zinc over time, with consequent negative cellular response. To overcome this unexpected result, a novel functionalization strategy requiring aqueous solution and mild conditions was designed. Sr/Mg-doped HT functionalized with this second strategy (via aldehyde peptide) showed a dramatic increase in human osteoblast proliferation at 6 days compared to only silanized or non-functionalized samples. Furthermore, we demonstrated that the functionalization treatment does not induce any cytotoxicity. Functionalized foams enhanced mRNA-specific transcript levels coding IBSP, VTN, RUNX2, and SPP1 at 2 days post-seeding. In conclusion, the second functionalization strategy proved to be appropriate for this specific biomaterial and was effective at enhancing the material's bioactivity.

In vitro evaluation of granules obtained from 3D sphene scaffolds and bovine bone grafts: chemical and biological assays.

Sphene is an innovative bone graft material. The aim of this study was to investigate and compare the physicochemical and biological properties of Bio-Oss® (BO) and in-lab synthesized and processed sphene granules. BO granules of 1000-2000 µm (BO-L), 250-1000 µm (BO-S) and 100-200 µm (BO-p) for derived granules, and corresponding groups of sphene granules obtained from 3D printed blocks (SB-L, SB-S, SB-p) and foams (SF-L, SF-S and SF-p) were investigated. The following analyses were conducted: morphological analysis, specific surface area and porosity, inductively coupled plasma mass spectrometry (ICP-MS), cytotoxicity assay, Alizarin staining, bone-related gene expression, osteoblast migration and proliferation assays. All pulverized granules exhibited a similar morphology and SF-S resembled natural bone. Sphene-derived granules showed absence of micro- and mesopores and a low specific surface area. ICP-MS revealed a tendency for absorption of Ca and P for all BO samples, while sphene granules demonstrated a release of Ca. No cellular cytotoxicity was detected and osteoblastic phenotype in primary cells was observed, with significantly increased values for SF-L, SF-S, BO-L and BO-p. Further investigations are needed before clinical use can be considered.

Advanced Dye Sorbents from Combined Stereolithography 3D Printing and Alkali Activation of Pharmaceutical Glass Waste.

Additive manufacturing (AM) technologies enable the fabrication of objects with complex geometries in much simpler ways than conventional shaping methods. With the fabrication of recyclable filters for contaminated waters, the present work aims at exploiting such features as an opportunity to reuse glass from discarded pharmaceutical containers. Masked stereolithography-printed scaffolds were first heat-treated at relatively low temperatures (680 and 730 °C for 1 h) and then functionalized by alkali activation, with the formation of zeolite and sodium carbonate phases, which worked as additional adsorbing centers. As-sintered and activated scaffolds were characterized in terms of the efficiency of filtration and removal of methylene blue, used as a reference dye. The adsorption efficiency of activated printed glass was 81%. The 3D-printed adsorbent can be easily separated from the solution for reuse.

Upcycling of Pharmaceutical Glass into Highly Porous Ceramics: From Foams to Membranes.

The present COVID-19 emergency has dramatically increased the demand for pharmaceutical containers, especially vials. End-of-life containers, however, cannot be easily recycled in the manufacturing of new articles. This paper presents some strategies for upcycling of pharmaceutical glass into various porous ceramics. Suspensions of a fine glass powder (70 vol%) are used as a starting material. Highly uniform cellular structures may be easily prepared by vigorous mechanical stirring of partially gelified suspensions with added surfactant, followed by drying and firing at 550-650 °C. Stabilization of the cellular structures at temperatures as low as the glass transition temperature (Tg) of the used glass is facilitated by thermal decomposition of the gel phase, instead of viscous flow sintering of glass. This finding enabled the preparation of glass membranes (∼78 vol% open porosity), by direct firing of hardened suspensions, avoiding any surfactant addition and mechanical stirring. The powders obtained by crushing of hardened suspensions, even in unfired state, may be used as a low-cost sorbent for dye removal.

Novel Functional Glass-Ceramic Coatings on Titanium Substrates from Glass Powders and Reactive Silicone Binders.

'Silica-defective glasses', combined with a silicone binder, have been already shown as a promising solution for the manufacturing of glass-ceramics with complex geometries. A fundamental advantage is the fact that, after holding glass powders together from room temperature up to the firing temperature, the binder does not completely disappear. More precisely, it converts into silica when heat-treated in air. A specified 'target' glass-ceramic formulation results from the interaction between glass powders and the binder-derived silica. The present paper is dedicated to the extension of the approach to the coating of titanium substrates (to be used for dental and orthopedic applications), with a bioactive wollastonite-diopside glass-ceramic layer, by the simple airbrushing of suspensions of glass powders in alcoholic silicone solutions. The interaction between glass and silica from the decomposition of the binder led to crack-free glass-ceramic coatings, upon firing in air; in argon, the glass/silicone mixtures yielded novel composite coatings, embedding pyrolytic carbon. The latter phase enabled the absorption of infrared radiation from the coating, which is useful for disinfection purposes.

Robocasting of advanced ceramics: ink optimization and protocol to predict the printing parameters - A review.

Direct-Ink-Writing (or robocasting) is a subset of extrusion-based additive manufacturing techniques that has grown significantly in recent years to design simple to complex ceramic structures. Robocasting, relies on the use of high-concentration powder pastes, also known as inks. A successful optimization of ink rheology and formulation constitutes the major key factor to ensure printability for the fabrication of self-supporting ceramic structures with a very precise dimensional resolution. However, to date achieving a real balance between a comprehensive optimization of ink rheology and the determination of a relevant protocol to predict the printing parameters for a given ink is still relatively scarce and has been not yet standardized in the literature. The current review reports, in its first part, a detailed survey of recent studies on how ink constituents and composition affect the direct-ink-writing of ceramic parts, taking into account innovative ceramic-based-inks formulations and processing techniques. Precisely, the review elaborates the major factors influencing on ink rheology and printability, specifically binder type, particle physical features (size, morphology and density) and ceramic feedstock content. In the second part, this review suggests a standardized guideline to effectively adapt a suitable setting of the printing parameters, such as printing speed and pressure, printing substrate, strut spacing, layer height, nozzle diameter in function of ink intrinsic rheology.

3D Printing of Hierarchically Porous Lattice Structures Based on Åkermanite Glass Microspheres and Reactive Silicone Binder.

The present study illustrates the manufacturing method of hierarchically porous 3D scaffolds based on åkermanite as a promising bioceramic for stereolithography. The macroporosity was designed by implementing 3D models corresponding to different lattice structures (cubic, diamond, Kelvin, and Kagome). To obtain micro-scale porosity, flame synthesized glass microbeads with 10 wt% of silicone resins were utilized to fabricate green scaffolds, later converted into targeted bioceramic phase by firing at 1100 °C in air. No chemical reaction between the glass microspheres, crystallizing into åkermanite, and silica deriving from silicone oxidation was observed upon heat treatment. Silica acted as a binder between the adjacent microspheres, enhancing the creation of microporosity, as documented by XRD, and SEM coupled with EDX analysis. The formation of 'spongy' struts was confirmed by infiltration with Rhodamine B solution. The compressive strength of the sintered porous scaffolds was up to 0.7 MPa with the porosity of 68-84%.

Polymer-Derived Biosilicate®-like Glass-Ceramics: Engineering of Formulations and Additive Manufacturing of Three-Dimensional Scaffolds.

Silicone resins, filled with phosphates and other oxide fillers, yield upon firing in air at 1100 °C, a product resembling Biosilicate® glass-ceramics, one of the most promising systems for tissue engineering applications. The process requires no preliminary synthesis of parent glass, and the polymer route enables the application of direct ink writing (DIW) of silicone-based mixtures, for the manufacturing of reticulated scaffolds at room temperature. The thermal treatment is later applied for the conversion into ceramic scaffolds. The present paper further elucidates the flexibility of the approach. Changes in the reference silicone and firing atmosphere (from air to nitrogen) were studied to obtain functional composite biomaterials featuring a carbon phase embedded in a Biosilicate®-like matrix. The microstructure was further modified either through a controlled gas release at a low temperature, or by the revision of the adopted additive manufacturing technology (from DIW to digital light processing).

Up-Cycling of LCD Glass by Additive Manufacturing of Porous Translucent Glass Scaffolds.

Additive manufacturing technologies, compared to conventional shaping methods, offer great opportunities in design versatility, for the manufacturing of highly porous ceramic components. However, the application to glass powders, later subjected to viscous flow sintering, involves significant challenges, especially in shape retention and in the achievement of a substantial degree of translucency in the final products. The present paper disclosed the potential of glass recovered from liquid crystal displays (LCD) for the manufacturing of highly porous scaffolds by direct ink writing and masked stereolithography of fine powders mixed with suitable organic additives, and sintered at 950 °C, for 1-1.5 h, in air. The specific glass, featuring a relatively high transition temperature (Tg~700 °C), allowed for the complete burn-out of organics before viscous flow sintering could take place; in addition, translucency was favored by the successful removal of porosity in the struts and by the resistance of the used glass to crystallization.

Biofunctionalization of bioactive ceramic scaffolds to increase the cell response for bone regeneration.

Biofunctionalization was investigated for polymers and metals considering their scarce integration ability. On the contrary few studies dealt with ceramic biofunctionalization because the bioactive and bioresorbable surfaces of ceramics are able to positively interact with biological environment. In this study the cell-response improvement on biofunctionalized wollastonite and diopside-based scaffolds was demonstrated. The ceramics were first obtained by heat treatment of a silicone embedding reactive oxide fillers and then biofunctionalized with adhesive peptides mapped on vitronectin. The most promisingin vitroresults, in terms of h-osteoblast proliferation and bone-related gene expression, were reached anchoring selectively a peptide stable toward proteolytic degradation induced by serum-enriched medium. Inin vivoassays the anchoring of this protease-stable adhesive peptide was combined with self-assembling peptides, for increasing cell viability and angiogenesis. The results demonstrated external and internal cell colonization of biofunctionalized scaffolds with formation of new blood vessels (neoangiogenesis) and stimulation of ectopic mineralization.

Influence of the thermal treatment on the characteristics of porous geopolymers as potential biomaterials.

Highly porous sodium geopolymer structures were successfully produced through the chemical direct foaming approach at ambient temperature. The impact of the thermal treatment, as well as the influence of various additions of hydrogen peroxide, as a foaming agent, on the porosity, microstructure and mechanical characteristics of the produced geopolymers was investigated. The evaluation of bioactivity was carried out by assessing the formation of an apatite layer on the samples' surface, using scanning electronic microscopy and inductively coupled plasma spectrometry for the simulated body fluid solution, in which the geopolymer samples were kept up to 28 days. In addition, the biodegradability was estimated through the weight change of the samples and pH-measurements. The results demonstrated that the geopolymer foams, produced using 4.5 vol% H2O2 and heat-treated at 500 °C for 1 h, possessed a high open porosity (71 vol %), excellent compressive strength (3.56 ± 0.27 MPa), and suitable chemical stability. The pH value of SBF solutions, in which these geopolymers were immersed for 28 days, remained close to the physiological one. The in vitro study indicated that the developed geopolymer foams possessed bioactivity, as demonstrated by the formation of apatite particles on their surface after immersion in the simulated body fluid solution for 28 days.

In vitro and in vivo study of naturally derived alginate/hydroxyapatite bio composite scaffolds.

Biogenic bioceramics scaffolds are receiving considerable attention for bone restoration applications. Compared with scaffolds of chemical origin, biogenic scaffolds exhibit greater biocompatibility and enhanced bioactive features. In the present study, porous biogenic hydroxyapatite (bHA) was prepared via a polymeric infiltration route and was subsequently coated with alginate to produce alginate/biogenic hydroxyapatite (Alg/bHA) composites. Alginate was used to enhance the mechanical properties as well as the bioactivity and biodegradability of the HA scaffolds. A coating of 3%w/v alginate applied for 10 min was found to result in the best coating for the HA porous scaffolds. The in vitro study demonstrated that the prepared composites had acceptable bioactivity and biodegradability characteristics. The histological study in femur bone of rats indicated that the 3Alg/HA scaffolds capable of supporting both endochondral and intramembranous bone formation. The defect was fully regenerated and mostly filled with the mature lamellar bone after 6 months, with Ca/P atomic ratio similar to the rat's normal bone. The studied scaffolds provide a promising therapeutic option to enhance local bone healing because they do not damage liver or kidney functions and do not induce carcinogenic or inflammatory effects. Accordingly, 3Alg/HA scaffolds are recommended for the tissue engineering applications.

An In Vivo Study in Rat Femurs of Bioactive Silicate Coatings on Titanium Dental Implants.

Silica-based ceramics have been proposed for coating purposes to enhance dental and orthopedic titanium (Ti) implant bioactivity. The aim of this study was to investigate the influence of sphene-based bioceramic (CaO.TiO2.SiO2) coatings on implant osseointegration in vivo. Sphene coatings were obtained from preceramic polymers and nano-sized active precursors and deposited by an automatic airbrush. Twenty customized Ti implants, ten sphene-coated and ten uncoated rough implants were implanted into the proximal femurs of ten Sprague-Dawley rats. Overall, cortical and cancellous bone-to-implant contact (BIC) were determined using micro-computed tomography (micro-CT) at 14 and 28 days. Moreover, peri-implant bone healing was histologically and histomorphometrically evaluated. The white blood cell count in the synovial fluid of the knee joints, if present, was also assessed. No difference in the BIC values was observed between the sphene-coated and uncoated implants, overall and in the two bone compartments (p > 0.05). Delamination of the coating occurred in three cases. Consistently with micro-CT data, the histological evaluation revealed no differences between the two groups. In addition, no synovial fluid could be collected on the test side, thus confirming sphene biocompatibility. In conclusion, sphene coating was found to be a suitable material for biomedical applications. Further studies are needed to improve coating adhesion to the implants.

Bioactive Glass and Silicate-Based Ceramic Coatings on Metallic Implants: Open Challenge or Outdated Topic?

The overall success and long-term life of the medical implants are decisively based on the convenient osseointegration at the hosting tissue-implant interface. Therefore, various surface modifications and different coating approaches have been utilized to the implants to enhance the bone formation and speed up the interaction with the surrounding hosting tissues, thereby enabling the successful fixation of implants. In this review, we will briefly present the main metallic implants and discuss their biocompatibility and osseointegration ability depending on their chemical and mechanical properties. In addition, as the main goal of this review, we explore the main properties of bioactive glasses and silica-based ceramics that are used as coating materials for both orthopedic and dental implants. The current review provides an overview of these bioactive coatings, with a particular emphasis on deposition methods, coating adhesion to the substrates and apatite formation ability tested by immersion in Simulated Body Fluid (SBF). In vitro and in vivo performances in terms of biocompatibility, biodegradability and improved osseointegration are examined as well.

Advanced Open-Celled Structures from Low-Temperature Sintering of a Crystallization-Resistant Bioactive Glass.

Most materials for bone tissue engineering are in form of highly porous open-celled components (porosity >70%) developed by means of an adequate coupling of formulations and manufacturing technologies. This paper is dedicated to porous components from BGMS10 bioactive glass, originally designed to undergo viscous flow sintering without crystallization, which is generally known to degrade the bioactivity of 45S5 bioglass. The adopted manufacturing technologies were specifically conceived to avoid any contamination and give excellent control on the microstructures by simple operations. More precisely, 'green' components were obtained by digital light processing and direct foaming of glass powders suspended in a photosensitive organic binder or in an aqueous solution, activated with an organic base, respectively. Owing to characteristic quite large sintering window of BGMS10 glass, sintering at 750 °C caused the consolidation of the structures generated at room temperature, without any evidence of viscous collapse.

Highly Porous Polymer-Derived Bioceramics Based on a Complex Hardystonite Solid Solution.

Highly porous bioceramics, based on a complex hardystonite solid solution, were developed from silicone resins and micro-sized oxide fillers fired in air at 950 °C. Besides CaO, SrO, MgO, and ZnO precursors, and the commercial embedded silicone resins, calcium borate was essential in providing the liquid phase upon firing and favouring the formation of an unprecedented hardystonite solid solution, corresponding to the formula (Ca0.70Sr0.30)2(Zn0.72Mg0.15Si0.13) (Si0.85B0.15)2O7. Silicone-filler mixtures could be used in the form of thick pastes for direct ink writing of reticulated scaffolds or for direct foaming. The latter shaping option benefited from the use of hydrated calcium borate, which underwent dehydration, with water vapour release, at a low temperature (420 °C). Both scaffolds and foams confirmed the already-obtained phase assemblage, after firing, and exhibited remarkable strength-to-density ratios. Finally, preliminary cell tests excluded any cytotoxicity that could be derived from the formation of a boro-silicate glassy phase.