Manufacturing of 3D-Printed Hybrid Scaffolds with Polyelectrolyte Multilayer Coating in Static and Dynamic Culture Conditions.

Three-dimensional printing (3DP) has emerged as a promising method for creating intricate scaffold designs. This study assessed three 3DP scaffold designs fabricated using biodegradable poly(lactic) acid (PLA) through fused deposition modelling (FDM): mesh, two channels (2C), and four channels (4C). To address the limitations of PLA, such as hydrophobic properties and poor cell attachment, a post-fabrication modification technique employing Polyelectrolyte Multilayers (PEMs) coating was implemented. The scaffolds underwent aminolysis followed by coating with SiCHA nanopowders dispersed in hyaluronic acid and collagen type I, and finally crosslinked the outermost coated layers with EDC/NHS solution to complete the hybrid scaffold production. The study employed rotating wall vessels (RWVs) to investigate how simulating microgravity affects cell proliferation and differentiation. Human mesenchymal stem cells (hMSCs) cultured on these scaffolds using proliferation medium (PM) and osteogenic media (OM), subjected to static (TCP) and dynamic (RWVs) conditions for 21 days, revealed superior performance of 4C hybrid scaffolds, particularly in OM. Compared to commercial hydroxyapatite scaffolds, these hybrid scaffolds demonstrated enhanced cell activity and survival. The pre-vascularisation concept on 4C hybrid scaffolds showed the proliferation of both HUVECs and hMSCs throughout the scaffolds, with a positive expression of osteogenic and angiogenic markers at the early stages.

Comparative retrieval analysis of contemporary mobile and fixed unicompartmental knee bearing designs.

There are two different concepts behind Unicompartmental Knee Replacement (UKR). Mobile bearings, as exemplified by the Oxford UKR, and fixed bearings, as exemplified by the Physica ZUK. These are the two most commonly implanted UKRs in the UK. For the first time, a comparison of the tribological features of 19 explanted Oxford and 19 explanted Physica ZUK UKRs was undertaken. Surface damage on the polyethylene (PE) inserts of the Oxford and Physica ZUK cohorts were assessed using an established semi-quantitative scoring method. The femoral components of both cohorts were assessed using a non-contact 3D profilometer to measure roughness values. It was found that the PE inserts of the Oxford cohort (22.54 ± 9.14) had statistically significant greater damage scores than the Physica ZUK cohort (16.50 ± 5.17) (p = 0.04). However, the femoral components of the Oxford cohort showed lower roughness values than the Physica ZUK cohort (p = 0.00). This is the first study that reports a comparative roughness analysis between retrieved Oxford and Physica ZUK UKR designs.

Self-crystallisation, an unexpected property of 45S5 Bioglass®.

Self-crystallisation of 45S5 Bioglass® powder and scaffolds was observed one year after their fabrication. Plate- and acicular-shape crystals, identified as calcium and sodium carbonates, grew at room temperature and atmospheric pressure, without any further treatment.

Mechanical properties of bioactive glasses, ceramics, glass-ceramics and composites: State-of-the-art review and future challenges.

The repair and restoration of bone defects in orthopaedic and dental surgery remains a major challenge despite advances in surgical procedures and post-operative treatments. Bioactive glasses, ceramics, glass-ceramics and composites show considerable potential for such applications as they can promote bone tissue regeneration. This paper presents an overview of the mechanical properties of various bioactive materials, which have the potential for bone regeneration. It also identifies current strategies for improving the mechanical properties of these novel materials, as these are rarely ideal as direct replacements for human bone. For this reason bioactive organic-inorganic composites and hybrids that have tailorable mechanical properties are of particular interest. The inorganic component (bioactive glass, ceramic or glass-ceramic) can provide both strength and bioactivity, while the organic component can add structural reinforcement, toughness and processability. Another topic presented in this paper includes 3D porous scaffolds that act as a template for cell attachment, proliferation and bone growth. Mechanical limitations of existing glass and ceramic scaffolds are discussed, along with the relevant challenges and strategies for further improvement. Advantages and disadvantages of different bioactive materials are critically examined. This paper is focused on optimization of biomaterials properties, in particular mechanical properties and bioactivity.

Chitosan based polymer/bioglass composites for tissue engineering applications.

Composite scaffolds formed from polymers and bioglasses have been widely explored for applications in regenerative medicine as they have suitable organic/inorganic structures and properties similar to human hard tissue. Yet, these materials have only been used for non-load-bearing or low load-bearing purposes as they have limited mechanical strength while research is focused on improving their properties. One method of improving mechanical strength is by covalently bonding the organic and inorganic phases. This has been successfully achieved in Class ll hybrids which have covalent bonding between polymers and bioglasses. As well as improving mechanical strength, the chemical connection of the two phases results in simultaneous degradation. The currently available composite scaffolds use collagen for the polymer phase which can cause allergic reactions and transmit pathogens. An alternative natural polymer is chitosan which has been used to create scaffolds with bioglass avoiding the issues arising from collagen. Additionally, using cross-linking agents has been shown to strengthen chitosan hydrogels improving their mechanical properties. A promising natural cross-linker is genipin which has lower toxicity than other cross-linking agents while producing hydrogels with improved mechanical properties compared to pure chitosan. In this paper we offer an overview of requirements, structures and currently available composite scaffolds for tissue engineering applications. We discuss the limitations of the currently available materials and consider the potential of covalently bonded hybrids particularly in relation to chitosan-based materials and the added benefits of genipin cross-linking.

Glass-ceramics for cancer treatment: So close, or yet so far?

After years of research on the ability of glass-ceramics in bone regeneration, this family of biomaterials has shown revolutionary potentials in a couple of emerging applications such as cancer treatment. Although glass-ceramics have not yet reached their actual potential in cancer therapy, the relevant research activity is significantly growing in this field. It has been projected that this idea and the advent of magnetic bioactive glass-ceramics and mesoporous bioactive glasses could result in major future developments in the field of cancer. Undoubtedly, this strategy needs further developments to better answer the critical questions essential for clinical usage. This review aims to address the existing research developments on glass-ceramics for cancer treatment, starting with the current status and moving to future advances. STATEMENT OF SIGNIFICANCE: Although glass-ceramics have not yet reached their potential in cancer therapy, research activity is significantly growing. It has been speculated that this idea and the advent of modern glass-ceramics could result in significant future advances. Undoubtedly, this strategy needs further investigations and many critical questions have to be answered before it can be successfully applied for cancer treatment. This paper reviews the current state-of-the-art, starting with current products and moving onto recent developments in this field. According to our knowledge, there is a lack of a systematic review on the importance and developments of magnetic bioactive glass-ceramics and mesoporous bioactive glasses for cancer treatment, and it is expected that this review will be of interest to those working in this area.

Osseointegration of porous apatite-wollastonite and poly(lactic acid) composite structures created using 3D printing techniques.

A novel apatite-wollastonite/poly(lactic acid) (AW/PLA) composite structure, which matches cortical and cancellous bone properties has been produced and evaluated in vitro and in vivo. The composites structure has been produced using an innovative combination of 3D printed polymer and ceramic macrostructures, thermally bonded to create a hybrid composite structure. In vitro cell assays demonstrated that the AW structure alone, PLA structure alone, and AW/PLA composite were all biocompatible, with the AW structure supporting the proliferation and osteogenic differentiation of rat bone marrow stromal cells. Within a rat calvarial defect model the AW material showed excellent osseointegration with the formation of new bone, and vascularisation of the porous AW structure, both when the AW was implanted alone and when it was part of the AW/PLA composite structure. However, the AW/PLA structure showed the largest amount of the newly formed bone in vivo, an effect which is considered to be a result of the presence of the osteoinductive AW structure stimulating bone growth in the larger pores of the adjacent PLA structure. The layered AW/PLA structure showed no signs of delamination in any of the in vitro or in vivo studies, a result which is attributed to good initial bonding between polymer and ceramic, slow resorption rates of the two materials, and excellent osseointegration. It is concluded that macro-scale composites offer an alternative route to the fabrication of bioactive bone implants which can provide a match to both cortical and cancellous bone properties over millimetre length scales.

Sensitivity of novel silicate and borate-based glass structures on in vitro bioactivity and degradation behaviour.

Three novel glass compositions, identified as NCL2 (SiO2-based), NCL4 (B2O3-based) and NCL7 (SiO2-based), along with apatite-wollastonite (AW) were processed to form sintered dense pellets, and subsequently evaluated for their in vitro bioactive potential, resulting physico-chemical properties and degradation rate. Microstructural analysis showed the carbonated hydroxyapatite (HCA) precipitate morphology following SBF testing to be composition-dependent. AW and the NCL7 formulation exhibited greater HCA precursor formation than the NCL2 and NCL4-derived pellets. Moreover, the NCL4 borate-based samples showed the highest biodegradation rate; with silicate-derived structures displaying the lowest weight loss after SBF immersion. The results of this study suggested that glass composition has significant influence on apatite-forming ability and also degradation rate, indicating the possibility to customise the properties of this class of materials towards the bone repair and regeneration process.

Novel bioglasses for bone tissue repair and regeneration: Effect of glass design on sintering ability, ion release and biocompatibility.

Eight novel silicate, phosphate and borate glass compositions (coded as NCLx, where x = 1 to 8), containing different oxides (i.e. MgO, MnO2, Al2O3, CaF2, Fe2O3, ZnO, CuO, Cr2O3) were designed and evaluated alongside apatite-wollastonite (used as comparison material), as potential biomaterials for bone tissue repair and regeneration. Glass frits of all the formulations were processed to have particle sizes under 53 µm, with their morphology and dimensions subsequently investigated by scanning electron microscopy (SEM). In order to establish the nature of the raw glass powders, X-ray diffraction (XRD) analysis was also performed. The sintering ability of the novel materials was determined by using hot stage microscopy (HSM). Ionic release potential was assessed by inductively coupled plasma optical emission spectroscopy (ICP-OES). Finally, the cytotoxic effect of the novel glass powders was evaluated for different glass concentrations via a colorimetric assay, on which basis three formulations are considered promising biomaterials.

Polyelectrolyte multi-layers assembly of SiCHA nanopowders and collagen type I on aminolysed PLA films to enhance cell-material interactions.

This paper presents a new approach in assembling bone extracellular matrix components onto PLA films, and investigates the most favourable environment which can be created using the technique for cell-material interactions. Poly (lactic acid) (PLA) films were chemically modified by covalently binding the poly(ethylene imine) (PEI) as to prepare the substrate for immobilization of polyelectrolyte multilayers (PEMs) coating. Negatively charged polyelectrolyte consists of well-dispersed silicon-carbonated hydroxyapatite (SiCHA) nanopowders in hyaluronic acid (Hya) was deposited onto the modified PLA films followed by SiCHA in collagen type I as the positively charged polyelectrolyte. The outermost layer was finally cross-linked by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrocholoride and N-hydroxysulfosuccinimide sodium salt (EDC/NHS) solutions. The physicochemical features of the coated PLA films were monitored via X-ray Photoelectron Spectroscopy (XPS) and Atomic Force Microscope (AFM). The amounts of calcium and collagen deposited on the surface were qualitatively and quantitatively determined. The surface characterizations suggested that 5-BL has the optimum surface roughness and highest amounts of calcium and collagen depositions among tested films. In vitro human mesenchymal stem cells (hMSCs) cultured on the coated PLA films confirmed that the coating materials greatly improved cell attachment and survival compared to unmodified PLA films. The cell viability, cell proliferation and Alkaline Phosphatase (ALP) expression on 5-BL were found to be the most favourable of the tested films. Hence, this newly developed coating materials assembly could contribute to the improvement of the bioactivity of polymeric materials and structures aimed to bone tissue engineering applications.

Development of multisubstituted hydroxyapatite nanopowders as biomedical materials for bone tissue engineering applications.

Ionic substitutions have been proposed as a tool to control the functional behavior of synthetic hydroxyapatite (HA), particularly for Bone Tissue Engineering applications. The effect of simultaneous substitution of different levels of carbonate (CO3 ) and silicon (Si) ions in the HA lattice was investigated. Furthermore, human bone marrow-derived mesenchymal stem cells (hMSCs) were cultured on multi-substituted HA (SiCHA) to determine if biomimetic chemical compositions were osteoconductive. Of the four different compositions investigates, SiCHA-1 (0.58 wt % Si) and SiCHA-2 (0.45 wt % Si) showed missing bands for CO3 and Si using FTIR analysis, indicating competition for occupation of the phosphate site in the HA lattice; 500°C was considered the most favorable calcination temperature as: (i) the powders produced possessed a similar amount of CO3 (2-8 wt %) and Si (<1.0 wt %) as present in native bone; and (ii) there was a minimal loss of CO3 and Si from the HA structure to the surroundings during calcination. Higher Si content in SiCHA-1 led to lower cell viability and at most hindered proliferation, but no toxicity effect occurred. While, lower Si content in SiCHA-2 showed the highest ALP/DNA ratio after 21 days culture with hMSCs, indicating that the powder may stimulate osteogenic behavior to a greater extent than other powders. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1775-1785, 2017.

In vitro biocompatibility of a ferrimagnetic glass-ceramic for hyperthermia application.

Ferrimagnetic glass-ceramics containing magnetite crystals were developed for hyperthermia applications of solid neoplastic tissue. The present work is focused on in vitro evaluation of the biocompatibility of these materials, before and after soaking in a simulated body fluid (SBF). X-ray diffraction, scanning electron microscopy, atomic absorption spectrophotometry, X-ray photoelectron spectrometry and pH measurements were employed in glass-ceramic characterisation. The free-radical mediated reactivity of the glass-ceramic was evaluated by Electron Paramagnetic Resonance (EPR) spin trapping. Cell adhesion and proliferation tests were carried out by using 3T3 murine fibroblasts. Cytotoxicity was performed by qualitative evaluation of human bone osteosarcoma cells U2OS cell line. The results show that almost two times more 3T3 cells proliferated on the samples pre-treated in SBF, compared with the untreated specimens. Moreover a decrease of confluence was observed at 48 and 72h for U2OS cells exposed to the untreated glass-ceramic, while the powder suspensions of glass-ceramic pre-treated in SBF did not influence the cell morphology up to 72h of exposition. The untreated glass-ceramic exhibited Fenton-like reactivity, as well as reactivity towards formate molecule. After pre-treatment with SBF the reactivity towards formate was completely suppressed. The concentration of iron released into the SBF solution was below 0.1ppm at 37°C, during one month of soaking. The different in vitro behaviour of the samples before and after SBF treatment has been correlated to the bioactive glass-ceramic surface modifications as detected by morphological, structural and compositional analyses.

Three-dimensional printing of porous load-bearing bioceramic scaffolds.

This article reports on the use of the binder jetting three-dimensional printing process combined with sintering to process bioceramic materials to form micro- and macroporous three-dimensional structures. Three different glass-ceramic formulations, apatite-wollastonite and two silicate-based glasses, have been processed using this route to create porous structures which have Young's modulus equivalent to cortical bone and average bending strengths in the range 24-36 MPa. It is demonstrated that a range of macroporous geometries can be created with accuracies of ±0.25 mm over length scales up to 40 mm. Hot-stage microscopy is a valuable tool in the definition of processing parameters for the sintering step of the process. Overall, it is concluded that binder jetting followed by sintering offers a versatile process for the manufacture of load-bearing bioceramic components for bone replacement applications.

Synthesis and Characterisation of Reduced Graphene Oxide/Bismuth Composite for Electrodes in Electrochemical Energy Storage Devices.

A reduced graphene oxide/bismuth (rGO/Bi) composite was synthesized for the first time using a polyol process at a low reaction temperature and with a short reaction time (60 °C and 3 hours, respectively). The as-prepared sample is structured with 20-50 nm diameter bismuth particles distributed on the rGO sheets. The rGO/Bi composite displays a combination of capacitive and battery-like charge storage, achieving a specific capacity value of 773 C g-1 at a current density of 0.2 A g-1 when charged to 1 V. The material not only has good power density but also shows moderate stability in cycling tests with current densities as high as 5 A g-1 . The relatively high abundance and low price of bismuth make this rGO/Bi material a promising candidate for use in electrode materials in future energy storage devices.

Composite bone cements loaded with a bioactive and ferrimagnetic glass-ceramic. Part I: Morphological, mechanical and calorimetric characterization.

Hyperthermia is a technique for destroying cancer cells which involves the exposition of body's tissue to a controlled heat, normally between 41℃ and 46℃. It has been reported that ferro- or ferrimagnetic materials can heat locally, if they are placed (after being implanted) under an alternating magnetic field, damaging only tumoral cells and not the healthy ones. The power loss produced by the magnetic materials can be dissipated in the form of heat. This phenomenon has to be regulated in order to obtain a controlled temperature inside the tissues. The material that was produced and characterized in this work is composed of two phases: a polymethylmethacrylate (PMMA) matrix in which a ferrimagnetic biocompatible/bioactive glass ceramic is dispersed. This composite material is intended to be applied as bone filler for the hyperthermic treatment of bone tumors. The ferrimagnetic bioactive glass-ceramic belongs to the system SiO2-Na2O-CaO-P2O5-FeO-Fe2O3 and contains magnetite (FeO*Fe2O3) inside an amorphous bioactive residual phase. The composite material possesses structural, magnetic and bioactivity properties. The structural ones are conferred by PMMA which acts as filler for the bone defect or its damaged area. Bioactivity is conferred by the composition of the residual amorphous phase of the glass-ceramic and magnetic properties are conferred by magnetite crystals embedded in the bioactive glass-ceramic. The characterization involved the following tests: morphological and chemical characterization (scanning electron microscopy-energy dispersion spectrometry-micro computed tomography analysis), calorimetric tests and mechanical test (compression and flexural four point test). In vitro assessment of biological behavior will be the object of the part II of this work.

Novel resorbable glass-ceramic scaffolds for hard tissue engineering: from the parent phosphate glass to its bone-like macroporous derivatives.

One of the major challenges of hard tissue engineering research focuses on the development of scaffolds that can match the mechanical properties of the host bone and resorb at the same rate as the bone is repaired. The aim of this work was the synthesis and characterization of a resorbable phosphate glass, as well as its application for the fabrication of three dimensional (3-D) scaffolds for bone regeneration. The glass microstructure and behaviour upon heating were analysed by X-ray diffraction, differential scanning calorimetry and hot stage microscopy. The glass solubility was investigated according to relevant ISO standards using distilled water, simulated body fluid (SBF) and Tris-HCl as testing media. The glass underwent progressive dissolution over time in all three media but the formation of a hydroxyapatite-like layer was also observed on the samples soaked in SBF and Tris-HCl, which demonstrated the bioactivity of the material. The glass powder was used to fabricate 3-D macroporous bone-like glass-ceramic scaffolds by adopting polyethylene particles as pore formers: during thermal treatment, the polymer additive was removed and the sintering of glass particles was allowed. The obtained scaffolds exhibited high porosity (87 vol.%) and compressive strength around 1.5 MPa. After soaking for 4 months in SBF, the scaffolds mass loss was 76 wt.% and the pH of the solution did not exceed the 7.55 value, thereby remaining in a physiological range. The produced scaffolds, being resorbable, bioactive, architecturally similar to trabecular bone and exhibiting interesting mechanical properties, can be proposed as promising candidates for bone repair applications.

Optimization of composition, structure and mechanical strength of bioactive 3-D glass-ceramic scaffolds for bone substitution.

Fabrication of 3-D highly porous, bioactive, and mechanically competent scaffolds represents a significant challenge of bone tissue engineering. In this work, Bioglass®-derived glass-ceramic scaffolds actually fulfilling this complex set of requirements were successfully produced through the sponge replication method. Scaffold processing parameters and sintering treatment were carefully designed in order to obtain final porous bodies with pore content (porosity above 70 %vol), trabecular architecture and mechanical properties (compressive strength up to 3 MPa) analogous to those of the cancellous bone. Influence of the Bioglass® particles size on the structural and mechanical features of the sintered scaffolds was considered and discussed. Relationship between porosity and mechanical strength was investigated and modeled. Three-dimensional architecture, porosity, mechanical strength and in vitro bioactivity of the optimized Bioglass®-derived scaffolds were also compared to those of CEL2-based glass-ceramic scaffolds (CEL2 is an experimental bioactive glass originally developed by the authors at Politecnico di Torino) fabricated by the same processing technique, in an attempt at understanding the role of different bioactive glass composition on the major features of scaffolds prepared by the same method.

Resorbable glass-ceramic phosphate-based scaffolds for bone tissue engineering: synthesis, properties, and in vitro effects on human marrow stromal cells.

Highly porous bioresorbable glass-ceramic scaffolds were prepared via sponge replication method by using an open-cell polyurethane foam as a template and phosphate-based glass powders. The glass, belonging to the P2O5-SiO2-CaO-MgO-Na2O-K2O system, was synthesized by a melting-quenching route, ground, and sieved to obtain powders with a grain size of less than 30 µm. A slurry containing glass powders, polyvinyl alcohol, and water was prepared to coat the polymeric template. The removal of the polymer and the sintering of the glass powders were performed by a thermal treatment, in order to obtain an inorganic replica of the template structure. The structure and properties of the scaffold were investigated from structural, morphological, and mechanical viewpoints by means of X-ray diffraction, scanning electron microscopy, density measurements, image analysis, and compressive tests. The scaffolds exhibited a trabecular architecture that closely mimics the structure of a natural spongy bone. The solubility of the porous structures was assessed by soaking the samples in acellular simulated body fluid (SBF) and Tris-HCl for different time frames and then by assessing the scaffold weight loss. As far as the test in SBF is concerned, the nucleation of hydroxyapatite on the scaffold trabeculae demonstrates the bioactivity of the material. Biological tests were carried out using human bone marrow stromal cells to test the osteoconductivity of the material. The cells adhered to the scaffold struts and were metabolically active; it was found that cell differentiation over proliferation occurred. Therefore, the produced scaffolds, being biocompatible, bioactive, resorbable, and structurally similar to a spongy bone, can be proposed as interesting candidates for bone grafting.