Phosphate glass fibre scaffolds: Tailoring of the properties and enhancement of the bioactivity through mesoporous glass particles.

Novel bone glass fibre scaffolds were developed by thermally bonding phosphate glass fibres belonging to the P2O5-CaO-Na2O-SiO2-MgO-K2O-TiO2 system (TiPS2.5 glass). Scaffolds with fibres of 85 or 110µm diameter were fabricated, showing compressive strength in the range of 2-3.5MPa, comparable to that of the trabecular bone. The effect of different thermal treatments and fibre diameters and length on the final scaffold structure was investigated by means of micro-CT analysis. The change of the sintering time from 30 to 60min led to a decrease in the scaffold overall porosity from 58 to 21vol.% for the 85µm fibre scaffold and from 50 to 40vol.% when increasing the sintering temperature from 490 to 500°C for the 110µm fibre scaffold. The 85µm fibres resulted in an increase of the scaffold overall porosity, increased pore size and lower trabecular thickness; the use of different fibre diameters allowed the fabrication of a scaffold showing a porosity gradient. In order to impart bioactive properties to the scaffold, for the first time in the literature the introduction in these fibre scaffolds of a bioactive phase, a melt-derived bioactive glass (CEL2) powder or spray-dried mesoporous bioactive glass particles (SD-MBG) was investigated. The scaffold bioactivity was assessed through soaking in simulated body fluid. CEL2/glass fibre scaffold did not show promising results due to particle detachment from the fibres during soaking in simulated body fluid. Instead the use of mesoporous bioactive powders showed to be an effective way to impart bioactivity to the scaffold and could be further exploited in the future through the ability of mesoporous particles to act as systems for the controlled release of drugs.

Bioactivity and Mechanical Stability of 45S5 Bioactive Glass Scaffolds Based on Natural Marine Sponges.

Bioactive glass (BG) based scaffolds (45S5 BG composition) were developed by the replica technique using natural marine sponges as sacrificial templates. The resulting scaffolds were characterized by superior mechanical properties (compression strength up to 4 MPa) compared to conventional BG scaffolds prepared using polyurethane (PU) packaging foam as a template. This result was ascribed to a reduction of the total scaffold porosity without affecting the pore interconnectivity (>99%). It was demonstrated that the reduction of total porosity did not affect the bioactivity of the BG-based scaffolds, tested by immersion of scaffolds in simulated body fluid (SBF). After 1 day of immersion in SBF, a homogeneous CaP deposit on the surface of the scaffolds was formed, which evolved over time into carbonate hydroxyapatite (HCA). Moreover, the enhanced mechanical properties of these scaffolds were constant over time in SBF; after an initial reduction of the maximum compressive strength upon 7 days of immersion in SBF (to 1.2 ± 0.2 MPa), the strength values remained almost constant and higher than those of BG-based scaffolds prepared using PU foam (<0.05 MPa). Preliminary cell culture tests with Saos-2 osteoblast cell line, namely direct and indirect tests, demonstrated that no toxic residues remained from the natural marine sponge templates and that cells were able to proliferate on the scaffold surfaces.

Novel systems for tailored neurotrophic factor release based on hydrogel and resorbable glass hollow fibers.

A novel system for the release of neurotrophic factor into a nerve guidance channel (NGC) based on resorbable phosphate glass hollow fibers (50P2O5-30CaO-9Na2O-3SiO2-3MgO-2.5K2O-2.5TiO2 mol%) in combination with a genipin-crosslinked agar/gelatin hydrogel (A/G_GP) is proposed. No negative effect on the growth of neonatal olfactory bulb ensheathing cell line (NOBEC) as well as on the expression of pro- and anti-apoptotic proteins was measured in vitro in the presence of fiber dissolution products in the culture medium. For the release studies, fluorescein isothiocyanate-dextran (FD-20), taken as growth factor model molecule, was solubilized in different media and introduced into the fiber lumen exploiting the capillary action. The fibers were filled with i) FD-20/phosphate buffered saline (PBS) solution, ii) FD-20/hydrogel solution before gelation and iii) hydrogel before gelation, subsequently lyophilized and then filled with the FD-20/PBS solution. The different strategies used for the loading of the FD-20 into the fibers resulted in different release kinetics. A slower release was observed with the use of A/G_GP hydrogel. At last, poly(ε-caprolactone) (PCL) nerve guides containing the hollow fibers and the hydrogel have been fabricated.

Phosphate glass fibres and their role in neuronal polarization and axonal growth direction.

Phosphate glass fibres with composition 50P(2)O(5)-30CaO-9Na(2)O-3SiO(2)-3MgO-(5-x)K(2)O-xTiO(2)mol.% (x=0, 2.5, 5, respectively coded as TiPS(0), TiPS(2.5) and TiPS(5)) were drawn following the preform drawing approach. A 20-day solubility test in bi-distilled water was carried out on glass fibres with different compositions and diameters ranging between 25 and 82 µm. The results show that the glass composition, the initial fibre diameter and the thermal treatment are the main factors influencing the dissolution kinetics and that the fibres maintain their structural integrity and composition during dissolution. Biological tests were carried out on aligned TiPS(2.5) glass fibres using Neonatal Olfactory Bulb Ensheathing Cell Line (NOBEC) and Dorsal Root Ganglia (DRG) neurons. The fibres showed to be permissive substrates for cell adhesion and proliferation. The aligned configuration of the fibres seemed to provide a directional cue for growing axons of DRG neurons, which showed to sprout and grow long neurites along the fibre axis direction. These promising findings encourages further studies to evaluate the potential use of resorbable glass fibres (e.g.in combination with a nerve guidance tube) for the enhancement of the peripheral nerve healing with the role of supporting and guiding the cells involved in the nerve regeneration.

Effects of TiO2-containing phosphate glasses on solubility and in vitro biocompatibility.

Phosphate-based glasses with different amounts of titanium dioxide (TiO(2)), having the following molar composition 50P(2)O(5)-30CaO-9Na(2)O-3SiO(2)-3MgO-(5-x)K(2)O-xTiO(2), (where x = 0, 2.5, 5 mol %), were synthesised and characterized in terms of solubility (according to ISO 10993-14), and in vitro biocompatibility using human MG-63 osteoblast cells. Dissolution tests were carried out in Tris-HCl (pH 7.4) to simulate the physiological pH and in citric acid (pH 3.0) to simulate an acidic environment. The weight loss decreased with increasing TiO(2) content, a process further enhanced in acidic medium. TiO(2) reduced the pH changes usually caused by the dissolution products released. Cellular tests showed that all the glasses studied (0-5 mol % TiO(2)) and TiCl(4), used to investigate the biocompatibility of titanium ions, did not produce cytotoxic effects on human MG-63 osteoblasts for up to 5 days in culture. On the basis of these results, we suggest that TiO(2)-containing phosphate glasses could be promising substrates for bone tissue engineering applications.

Response of human bone marrow stromal cells to a resorbable P(2)O(5)-SiO(2)-CaO-MgO-Na(2)O-K(2)O phosphate glass ceramic for tissue engineering applications.

This work focuses on the synthesis and characterization of a novel bioresorbable glass ceramic phosphate-based material (GC-ICEL). More specifically, its solubility in different aqueous media (water, Tris-HCl and acellular simulated body fluid) and the response of human stromal cells cultured on it were investigated. X-ray diffraction analysis showed the presence of two crystalline phases identified as Na(2)Mg(PO(4))(3) and Ca(2)P(2)O(7) and dissolution tests highlighted a preferential dissolution of the Na(2)Mg(PO(4))(3) phase and of the residual amorphous phase in all the chosen media. Soaking tests in simulated body fluid showed precipitation of a hydroxyapatite layer, demonstrating the bioactivity of GC-ICEL, which is partially due to the reported bioactivity of Ca(2)P(2)O(7). The effect of GC-ICEL on adhesion, proliferation and osteoblastic gene expression of human bone marrow-derived stromal cells was also studied. Combining molecular and biochemical analyses, it was found that bone marrow cell differentiation was stimulated over proliferation on GC-ICEL. Moreover, the expression of bone-related genes in cells cultured on GC-ICEL confirmed the bioactivity of this phosphate-based glass ceramic, which might have a stimulatory effect on osteogenesis.