Porous and strong bioactive glass (13–93) scaffolds fabricated by freeze extrusion technique

Scaffolds fabricated by current methods often lack the combination of high strength and high porosity for skeletal substitution of load-bearing bones. In this work, freeze extrusion fabrication (FEF), a solid freeform fabrication technique, was investigated for the creation of porous and strong bioactive glass (13–93) scaffolds for potential applications in the repair of loaded bone. The process parameters for forming three-dimensional (3D) scaffolds with a pre-designed, grid-like microstructure by FEF were determined. Following thermal treatment of the as-formed constructs at temperatures up to 700 °C, scaffolds consisting of dense glass struts and interconnecting pores (porosity ≈ 50%; pore width ≈ 300 μm) were obtained. These scaffolds showed an elastic mechanical response in compression, with a compressive strength of 140 ± 70 MPa and an elastic modulus of 5.5 ± 0.5 GPa, comparable to the values for human cortical bone. The scaffolds supported the proliferation of osteogenic cells in vitro, showing their biocompatibility. These results indicate that 13–93 bioactive glass scaffolds created by the FEF method could have potential application in the repair and regeneration of load-bearing bones.     

Sol–gel-derived manganese-releasing bioactive glass as a therapeutic approach for bone tissue engineering

Sol–gel processing allows the production of bioactive glasses (BG) with flexible compositions and the incorporation of different metallic ions with therapeutic benefits into the glass network. Manganese is among several previously studied therapeutically beneficial ions and has been shown to favour osteogenic differentiation, in addition to playing an important role in cell adhesion. The incorporation of Mn into bioactive glasses for tissue engineering has been previously conducted using the conventional melting route, whereas the sol–gel route has not yet been explored. Sol–gel technology has great versatility, allowing the preparation of BG with various compositions, sizes, morphologies and a large surface area that could provide improved cellular responses and enhanced bioactivity when compared to melt-derived glasses. In this context, this work developed new compositions of sol–gel bioactive glasses (on the SiO₂–P₂O₅–CaO–MnO system) and explored the effects of incorporating MnO on the structure, texture, in vitro bioactivity and cytocompatibility of these materials. Our results show that Mn-containing bioactive glasses present an amorphous character, high surface area and mesoporous structure. The formation of a hydroxycarbonate apatite (HCA) layer after immersion in simulated body fluid (SBF) revealed the high bioactivity of the glasses. Ion release evaluation indicated that the Si, Ca, P and Mn release levels could be adjusted within therapeutic limits, and cytotoxic analysis demonstrated that the ionic products of all samples generated a cell-friendly environment. Therefore, Mn incorporation into the bioactive glass network appears to be a potential strategy to develop superior materials with sustained ion release for tissue engineering.     

3-D high-strength glass–ceramic scaffolds containing fluoroapatite for load-bearing bone portions replacement

This study aimed to fabricate and investigate the structure, mechanical properties and bioactivity of three-dimensional (3-D) glass–ceramic scaffolds for bone tissue engineering. The scaffold material was a fluoroapatite-containing glass–ceramic synthesized by a melting–quenching route. Glass–ceramic powders were mixed with polyethylene particles acting as pore formers; the blend was pressed to obtain “green” compacts that were thermally treated to remove the organic phase and to sinter the inorganic one. The structure and morphology of the resulting scaffolds were characterized by X-ray diffraction, scanning electron microscopy, density measurements and capillarity tests. Crushing tests were carried out to investigate the mechanical properties of the scaffolds. The in vitro bioactivity was assessed by soaking the scaffolds in simulated body fluid for different time frames and by analyzing the modifications that occurred on the sample surface. The scaffolds had an interconnected macroporous structure with pores up to 50% vol. and they showed an orthotropic mechanical behaviour and strength well above 20 MPa. In addition, in vitro tests put into evidence the excellent bioactivity of the material. Therefore, the prepared scaffolds can be used in bone reconstructive surgery as effective load-bearing grafts thanks to their ease of tailoring, bioactive properties and high mechanical strength.     

Dissolution characteristics of extrusion freeformed hydroxyapatite–tricalcium phosphate scaffolds

The dissolution behaviour of calcium phosphate filaments made by extrusion freeforming for hard tissue scaffolds was measured. The solubility of filaments with different HA/beta-TCP ratios sintered at temperatures from 1,100 to 1,300 degrees C was measured under simulated physiological conditions (tris buffer solution: tris(hydroxyl) methyl-aminomethane-HCl), pH 7.4, 37 degrees C). Calcium and phosphate concentrations were measured separately by inductively coupled plasma (ICP) atomic emission spectroscopy. Surface morphologies and composition before and after immersion were analyzed by SEM and EDS. The results clearly show that as the beta-TCP content increased, the dissolution increased. Higher sintering temperatures, with consequent closure of surface pores, resulted in lower dissolution. Examination of the surface suggested dissolution on preferred sites by pitting.     

Biocompatible glass–ceramic materials for bone substitution

A new bioactive glass composition (CEL2) in the SiO₂–P₂O₅–CaO–MgO–K₂O–Na₂O system was tailored to control pH variations due to ion leaching phenomena when the glass is in contact with physiological fluids. CEL2 was prepared by a traditional melting-quenching process obtaining slices that were heat-treated to obtain a glass-ceramic material (CEL2GC) that was characterized thorough SEM analysis. Pre-treatment of CEL2GC with SBF was found to enhance its biocompatibility, as assessed by in vitro tests. CEL2 powder was then used to synthesize macroporous glass–ceramic scaffolds. To this end, CEL2 powders were mixed with polyethylene particles within the 300–600 μm size-range and then pressed to obtain crack-free compacted powders (green). This was heat-treated to remove the organic phase and to sinter the inorganic phase, leaving a porous structure. The biomaterial thus obtained was characterized by X-ray diffraction, SEM equipped with EDS, density measurement, image analysis, mechanical testing and in vitro evaluation, and found to be a glass–ceramic macroporous scaffold with uniformly distributed and highly interconnected porosity. The extent and size-range of the porosity can be tailored by varying the amount and size of the polyethylene particles.     

Characterisation of the bioactive behaviour of sol–gel hydroxyapatite–CaO and hydroxyapatite–CaO–bioactive glass composites

The fabrication and characterization of sol–gel derived hydroxyapatite–calcium oxide (HAp–CaO) material is investigated focusing on the effect of the addition of a bioactive glass on the material bioactive behaviour through the fabrication of a novel HAp–CaO (70 wt.%)–bioactive glass (30 wt.%) composite material. The bioactive behaviour of the materials was assessed by immersion studies in Simulated Body Fluid (SBF) and the alterations of the materials surfaces after soaking periods in SBF were characterized by Scanning Electron Microscopy (SEM) and Fourier Transform Infrared Spectroscopy (FTIR). A brittle and weakly crystalline carbonate hydroxyapatite (HCAp) layer was found to develop on the surface of all samples, few hours after immersion in SBF, confirming the high bioactivity of the material. Alterations of the morphology of the developed HCAp layer, which led to a more compact structure, were observed on the surface of composite samples after 7 days of immersion in SBF. The presence of the CaO phase seems to accelerate the formation of HCAp, while the bioactive glass affects both the morphology and cohesion of the developed layer.     

Physical and cytocompatibility properties of bioactive glass–polyvinyl alcohol–sodium alginate biocomposite foams prepared via sol–gel processing for trabecular bone regeneration

In the present work, biocomposite foams of bioactive glass along with polyvinyl alcohol and sodium alginate are designed and developed as a potential biomaterial for bone regeneration. These biocomposite foams have a low density of 0.92 g/cm³, providing desired property for bone tissue engineering applications. Biocomposite foams were prepared via surfactant foaming. Scanning electron microscopic characterization revealed pore size of 200–500 μm of the biocomposite foams. When these materials were incubated in simulated body fluid, hydroxyapatite layer formation was observed on the material surface. To confirm the cell viability and proliferation on these materials, MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay was performed with NIH 3T3 fibroblast cells and the results revealed good biocompatibility with the biocomposite foams. Cell adhesion studies further confirmed the biocompatibility of the scaffolds via cell attachment and ECM production. The optimally synthesized biocomposite foams had a good combination of physical properties with compressive strength of 1.64 MPa and elastic modulus of 18 MPa. In view of the favorable combination of physical and biological properties, the newly developed materials are considered to be suitable for regeneration of trabecular bone.     

Evaluation of 3D nano–macro porous bioactive glass scaffold for hard tissue engineering

Recently, nano–macro dual-porous, three-dimensional (3D) glass structures were developed for use as bioscaffolds for hard tissue regeneration, but there have been concerns regarding the interconnectivity and homogeneity of nanopores in the scaffolds, as well as the cytotoxicity of the environment deep inside due to limited fluid access. Therefore, mercury porosimetry, nitrogen absorption, and TEM have been used to characterize nanopore network of the scaffolds. In parallel, viability of MG 63 human osteosarcoma cells seeded on scaffold surface was investigated by fluorescence, confocal and electron microscopy methods. The results show that cells attach, migrate and penetrate inside the glass scaffold with high proliferation and viability rate. Additionally, scaffolds were implanted under the skin of a male New Zealand rabbit for in vivo animal test. Initial observations show the formation of new tissue with blood vessels and collagen fibers deep inside the implanted scaffolds with no obvious inflammatory reaction. Thus, the new nano–macro dual-porous glass structure could be a promising bioscaffold for use in regenerative medicine and tissue engineering for bone regeneration.     

Hybrid composite scaffolds prepared by sol–gel method for bone regeneration

This study reports the synthesis and characterization of hydroxyapatite (HA)/polycaprolactone (PCL) hybrid composite materials synthesized by sol–gel method. The fabrication of scaffolds was performed by salt-leaching technique using NaCl as porogen agent. In the first step, the physico-chemical characterization of composite material was performed to evaluate the composition and the interaction between the organic/inorganic phases. In the second step, optimized scaffolds were bioactivated on the surface. The combined effect of scaffold morphology and surface bioactivity is ideal for bone tissue engineering, supporting bone cells adhesion, proliferation and differentiation. Here, a combined strategy involving biomimetic approach, using a supersaturated Simulated Body Fluid (SBF), and salt-leaching technique has been developed to grow hydroxyapatite in composite scaffolds able to regenerate the natural bone.     

Glass–ceramic scaffolds containing silica mesophases for bone grafting and drug delivery

Glass–ceramic macroporous scaffolds were prepared using glass powders and polyethylene (PE) particles of two different sizes. The starting glass, named as Fa-GC, belongs to the system SiO₂–P₂O₅–CaO–MgO–Na₂O–K₂O–CaF₂ and was synthesized by a traditional melting-quenching route. The glass was ground and sieved to obtain powders of specific size which were mixed with PE particles and then uniaxially pressed in order to obtain crack-free green samples. The compact of powders underwent a thermal treatment to remove the organic phase and to sinter the Fa-GC powders. Fa-GC scaffolds were characterized by means of X-Ray Diffraction, morphological observations, density measurements, image analysis, mechanical tests and in vitro tests. Composite systems were then prepared combining the drug uptake-delivery properties of MCM-41 silica micro/nanospheres with the Fa-GC scaffold. The system was prepared by soaking the scaffold into the MCM-41 synthesis batch. The composite scaffolds were characterized by means of X-Ray Diffraction, morphological observations, mechanical tests and in vitro tests. Ibuprofen was used as model drug for the uptake and delivery analysis of the composite system. In comparison with the MCM-41-free scaffold, both the adsorption capacity and the drug delivery behaviour were deeply affected by the presence of MCM-41 spheres inside the scaffold.     

Development,characterization, and in vitro bioactivity studies of sol–gel bioactive glass/poly(l-lactide) nanocomposite scaffolds

Developing materials combining the advantages of synthetic polymers and bioactive glass nanoparticles can provide an efficient bone engineering scaffold. In this study, sol–gel bioactive glass (SG) nanoparticles were synthesized by quick alkali-mediation; sol–gel derived bioactive glass/poly(l-lactide) nanocomposite scaffolds were then developed. The influence of the glass content on the porosity of nanocomposite scaffolds was evaluated by SEM. The results showed that the neat polymer scaffold (PLA) has a highly interconnected porous structure with a maximum pore size of about 250 μm. For the composite scaffold containing 25 wt.% glass (SGP25), the decrease in the maximum pore size, (to about 200 μm) was not significant while for the SGP50 composite scaffold containing 50 wt.% glass it was a significant decrease (to about 100 μm). The apparent porosity of the scaffolds was 56.56% ± 7.15, 54.14% ± 3.84, and 53.11% ± 3.99 for PLA, SGP25, and, SGP50 respectively. FT-IR, TGA, and XRD results revealed some interaction of the glass filler with the polymeric matrix in the scaffolds. The degradation study showed that, by increasing the glass content in the scaffolds, the water absorption decreased, the weight loss increased, and the cumulative ion concentrations released from them also increased. This indicates the possibility of modulating the degradation rate by varying the glass/polymer ratio. At the end of the incubation period, the weight losses were around 5.44% ± 0.96, 32.50% ± 2.73, and 41.47% ± 3.02 for the PLA, SGP25, and SGP50, respectively. Moreover, the water uptake reached 119.65% ± 18.88 and 93.39% ± 13.01 for SGP25 and SGP50, respectively. The addition of the SG to the scaffolds was found to enhance their in vitro bioactivity. Therefore, these nanocomposite scaffolds have a potential to be applied in bone engineering. All data are expressed as mean ± standard deviation (n = 3).     

Composite bone substitute materials based on β-tricalcium phosphate and magnesium-containing sol–gel derived bioactive glass

In the present study, bioceramic composites with improved mechanical and biological properties were synthesized by sintering mixtures of β-tricalcium phosphate and SiO₂–CaO–MgO–P₂O₅ sol–gel derived bioactive glass at 1000–1200°C. The physical, mechanical, structural and biological properties of the composites were evaluated by appropriate experiments such as microhardness, bending strength, XRD, SEM and MTT. The results showed that 1000 and 1100°C were not appropriate temperatures for sintering the composites and in contrast, the microhardness, bending strength and bulk density significantly increased by increasing in quantity of bioglass phase when the samples were sintered at 1200°C. No significant difference was found between the fracture toughness of the composites and pure β-tricalcium phosphate. β-tricalcium phosphate was structurally stable up to 1200°C and did not transform to its alpha form even in the presence of the bioglass phase but migration of magnesium cations from the glass composition into its lattice structure was found by right-shift in XRD patterns, especially when the composite contained higher amount of bioglass component. Calcium silicate was also crystallized in the composition of the composites, which was more detectable in higher sintering temperatures. The results of the MTT test showed that proliferation of human osteosarcoma cells on the composites was considerably better than that of pure β-TCP.     

Sol–gel derived composite from bioactive glass–polyvinyl alcohol

Investigation of novel biomaterials for bone engineering is based on the development of porous scaffolds, which should match the properties of the tissue that is to be replaced. These materials need to be biocompatible, ideally osteoinductive, osteoconductive, and mechanically well-matched. In the present paper, we report the preparation and characterization of hybrid macroporous scaffold of polyvinyl alcohol (PVA)/bioactive glass through the sol–gel route. Hybrids containing PVA (80, 70 and 60 wt%) and bioactive glass with composition 58SiO₂–33CaO–9P₂O₅ were synthesized by foaming a mixture of polymer solution and bioactive glass via sol–gel precursor solution. PVA with two different degree of hydrolysis (DH), 98.5% (high degree) and 80% (low degree) were also investigated, in order to evaluate the influence of residual acetate group present in polymer chain on the final structure and properties of 3D porous composite produced. The microstructure, morphology and crystallinity of the hybrid porous scaffolds were characterized by X-ray diffraction (XRD), Infrared Fourier Transform spectrometry (FTIR) and Scanning electron microscopy (SEM/EDX) analysis. In addition, specific surface area was assessed by B.E.T. nitrogen adsorption method and mechanical behavior was evaluated by compression tests. Preliminary cytotoxicity and cell viability were also performed by the MTT assay. VERO cell monolayers were grown in 96-well microtiter plates. The results have clearly showed that hybrid foams of polyvinyl alcohol/bioactive glass (PVA/BG) with interconnected macroporous 3D structure were successfully produced. All the tested hybrids of PVA/BG have showed adequate cell viability properties for potential biological applications.     

Use of extrusion aids for successful production of Kollidon® CL-SF pellets by extrusion–spheronization

Objective: Fine particle ethylcellulose (FPEC) or poly(ethylene oxide) (PEO) addition to a Kollidon CL-SF was investigated to address low yield and poor sphericity in extruded-spheronized pellets.

Significance: The success of crospovidone as a diluent in extrusion–spheronization was dependent on a small particle size of the polymer. FPEC aided production of rugged and spherical pellets using a large particle size grade, Polyplasdone^® XL. PEO acted as an extrusion–spheronization aid when ethylcellulose was the diluent. These extrusion–spheronization aids could serve in this role when Kollidon^® CL-SF (K CL-SF) is the diluent.

Methods: The influence of formulation and process variables on pellet properties was investigated using design of experiments. A planetary mixer was used to prepare powder blends and the wetted mass after addition of water. An EXD 60 extruder produced extrudate that was spheronized in a Q230 marumerizer. Wet pellets were dried in a forced-air oven.

Results: FPEC improved rounding up but reduced pellet yield. Poly(ethylene oxide) imparted desired characteristics to the wetted mass, the extrudate, and the spheronized pellets. Pellet average diameter, yield, sphericity, aspect ratio, friability, and dissolution profile were assessed. Equations for pellet characteristics facilitated discussion of the influences of factors and their interactions. Optimization was performed on pellets that included PEO.

Conclusions: PEO proved to be an exceptional extrusion–spheronization aid in the preparation of pellets using K CL-SF. It facilitated wetted mass extrusion with minimal mass loss to the extruder, and markedly improved the sphericity of the pellets produced by marumerization. Immediate release pellets were obtained.     

Sol–gel derived nanoscale bioactive glass (NBG) particles reinforced poly(ε-caprolactone) composites for bone tissue engineering

This study investigated the effect of the addition of sol–gel derived nanoscale bioactive glass (NBG) particles on the mechanical properties and biological performances of PCL polymer, in order to evaluate the potential applications of PCL/NBG composites for bone tissue regeneration. Regardless of the NBG contents (10, 20, and 30 wt.%), the NBG particles, which were synthesized through the sol–gel process using polyethylene glycol (PEG) polymer as a template, could be uniformly dispersed in the PCL matrix, while generating pores in the PCL/NBG composites. The elastic modulus of the PCL/NBG composites increased remarkably from 89 ± 11 MPa to 383 ± 50 MPa with increasing NBG content from 0 to 30 wt.%, while still showing good ultimate tensile strength in the range of 15–19 MPa. The hydrophilicity, water absorption and degradation behavior of the PCL/NBG composites were also enhanced by the addition of the NBG particles. Furthermore, the PCL/NBG composite with a NBG content of 30 wt.% showed significantly enhanced in vitro bioactivity and cellular response compared to those of the pure PCL.     

Biomimetic design and fabrication of porous chitosan–gelatin liver scaffolds with hierarchical channel network

The presence of a hierarchical channel network in tissue engineering scaffold is essential to construct metabolically demanding liver tissue with thick and complex structures. In this research, chitosan–gelatin (C/G) scaffolds with fine three-dimensional channels were fabricated using indirect solid freeform fabrication and freeze-drying techniques. Fabrication processes were studied to create predesigned hierarchical channel network inside C/G scaffolds and achieve desired porous structure. Static in-vitro cell culture test showed that HepG2 cells attached on both micro-pores and micro-channels in C/G scaffolds successfully. HepG2 proliferated at much higher rates on C/G scaffolds with channel network, compared with those without channels. This approach demonstrated a promising way to engineer liver scaffolds with hierarchical channel network, and may lead to the development of thick and complex liver tissue equivalent in the future.     

Biofunctionalization of porous titanium coatings through sol–gel impregnation with a bioactive glass–ceramic

In implant technology, open porous Ti coatings are applied as functional surface layers on prosthetic devices to improve osseointegration. Since a successful clinical performance strongly depends on the (initial) quality of bone ingrowth in the porous structure, surface functionalization of the porous Ti to incorporate an additional osteoconductive capacity is recommended. In this paper, a bioactive glass–ceramic coating is applied into the open porous network of Ti coatings with a pore throat size of 1–20 μm through a sol–gel process. Using an all-alkoxide precursor route, homogeneous amorphous powders of three- (SiO₂–CaO–P₂O₅) and four-component (SiO₂–CaO–Na₂O–P₂O₅) bioactive glass compositions are prepared. By sol impregnation followed by a heat treatment, it is possible to deposit a micrometer thin bioactive glass–ceramic layer on the walls of the internal pore surface, while the original porosity and the open pore structure of the Ti coatings are maintained. The tensile adhesion strength of the Ti/bioactive glass–ceramic composite coatings is 22 to 29 MPa, suggesting a good mechanical adhesion.     

Rheological evaluations and in vitro studies of injectable bioactive glass–polycaprolactone–sodium alginate composites

Composite pastes composed of various amounts of melt-derived bioactive glass 52S4 (MG5) and polycaprolactone (PCL) microspheres in sodium alginate solution were prepared. Rheological properties in both rotatory and oscillatory modes were evaluated. Injectability was measured as injection force versus piston displacement. In vitro calcium phosphate precipitation was also studied in simulated body fluid (SBF) and tracked using scanning electron microscopy, X-ray diffraction and FTIR analyses. All composite pastes were thixotropic in nature and exhibited shear thinning behavior. The magnitude of thixotropy decreased by adding 10–30 wt% PCL, while further amounts of PCL increased it again. Moreover, the composites were viscoelastic materials in which the elastic modulus was higher than viscous term. The pastes which were just made of MG5 or PCL had poor injectability, whereas the composites containing both of these constituents exhibited reasonable injectability. All pastes revealed adequate structural stability in contact with SBF solution. In vitro calcium phosphate precipitation was well observed on the paste made of MG5 and somewhat on the pastes with 10–40 wt% PCL, however the precipitated layer was amorphous in nature. Overall, the produced composites may be appropriate as injectable biomaterials for non-invasive surgeries but more biological evaluations are essential.     

Highly porous 45S5 bioglass-derived glass–ceramic scaffolds by gelcasting of foams

Several types of alloplastic (artificial) grafts, known as scaffolds, have been developed for the treatment of bone defects caused by trauma and/or infection. Among the materials used to manufacture scaffolds, 45S5 Bioglass is a bioceramic that arouses significant interest due to ease preparation and excellent bioactive response. Among the various processing methods cited in the literature for the production of bioactive glass scaffolds, gelcasting is a method that produces macroporous structures, with interconnected and spherical pores and high mechanical strength. However, in the literature there are few reports about bioactive glass scaffolds produced by gelcasting method. In this work, 45S5-BG scaffolds were produced by gelcasting of foams varying the amount of foaming agent in order to optimize the desirable characteristics of the scaffold. The scaffolds show porosity between 70 and 86% and compressive strength of 1.22?±?0.7 and 0.78?±?0.4 MPa. In the biological studies, all 45S5-BG scaffolds showed cytocompatibility towards human osteoblastic cells and bioactive properties using SBF assay.     

Evaluation of physical–chemical properties and biocompatibility of a microrough and smooth bioactive glass particles

The purpose of this study was to evaluate physical-chemical and biocompatibility characteristics of a simple synthesis and low cost experimental bioactive glass. Physical and chemical properties were analyzed using scanning electron microscopy (SEM), X-ray energy dispersive (EDX), X-ray fluorescence (XRF) and X-ray diffraction (XRD). The biomaterials were subcutaneously implanted into rats, according to the following groups: G1, PerioGlastrade mark; G2, Biograntrade mark, G3, Experimental Bioactive Glass U (BGU) and G4, Control (Sham). After 7, 15, 21, 45, and 60 days, 5 animals/group/period were sacrificed and the subcutaneous tissue was dissected for histological and histometric analysis, considering inflammatory reaction and granulation area, presence of polymorphonuclear (PMN), monuclear (MN) and fibroblast (F) cells. SEM analysis of biomaterials showed irregular particles with different surface characteristics. EDX showed calcium, oxygen, sodium, phosphorus and silicon; XRF revealed silica oxide (SiO(2)), sodium oxide (Na(2)O), calcium oxide (CaO) and phosphorus oxide (P(2)O(5)). XRD indicated non crystalline phase. Measurement of tissue reaction showed similar results among the experimental groups at 45 and 60 days. No difference was found for PMN, MN and F cell counts. All biomaterials exhibited partial resorption. In conclusion, the experimental bioactive glass analyzed showed physical and chemical characteristics similar to the commercially available biomaterials, and was considered biocompatible, being partially reabsorbed in the subcutaneous tissue.