Polymer-Based Honeycomb Films on Bioactive Glass: Toward a Biphasic Material for Bone Tissue Engineering Applications.

The development of innovative materials for bone tissue engineering to promote bone regeneration while avoiding fibrous tissue infiltration is of paramount importance. Here, we combined the known osteopromotive properties of bioactive glasses (BaGs) with the biodegradability, biocompatibility, and ease to shape/handle of poly-l-co-d,l-lactic acid (PLDLA) into a single biphasic material. The aim of this work was to unravel the role of the surface chemistry and topography of BaG surfaces on the stability of a PLDLA honeycomb membrane, in dry and wet conditions. The PLDLA honeycomb membrane was deposited using the breath figure method (BFM) on the surface of untreated BaG discs (S53P4 and 13-93B20), silanized with 3-aminopropyltriethoxysilane (APTES) or conditioned (immersed for 24 h in TRIS buffer solution). The PLDLA membranes deposited onto the BaG discs, regardless of their composition or surface treatments, exhibited a honeycomb-like structure with pore diameter ranging from 1 to 5 µm. The presence of positively charged amine groups (APTES grafting) or the precipitation of a CaP layer (conditioned) significantly improved the membrane resistance to shear as well as its stability upon immersion in the TRIS buffer solution. The obtained results demonstrated that the careful control of the substrate surface chemistry enabled the deposition of a stable honeycomb membrane at their surface. This constitutes a first step toward the development of new biphasic materials enabling osteostimulation (BaG) while preventing migration of fibrous tissue inside the bone defect (honeycomb polymer membrane).

Phosphate/oxyfluorophosphate glass crystallization and its impact on dissolution and cytotoxicity.

The role of fluorine in bioactive glasses is of interest due to the potential of precipitating fluorapatite, a phase with higher chemical resistance than the typical hydroxyapatite precipitated from oxide bioactive glasses. However, the introduction of fluorine in silicate bioactive glasses was found deleterious to the bioactivity of the glass. Here, phosphate glasses with the composition 75NaPO3-(25-x) CaO-xCaF2 (in mol%), with x = 0-20 and glass-ceramics were investigated to evaluate their potential as substitutes to the traditional silicate bioactive glass. An increase in CaF2 substitution for CaO led to an increase in the glass solubility, due to an increase in highly soluble F(M)n species (where M is a cation) and to an increased polymerization of the phosphate network. Structural analysis reveals the formation of FP bonds, in addition to the F(M)n species, in the glass with the higher CaF2 content. Furthermore, with heat treatment, CaF2 crystals precipitate within the bulk in the newly developed glass, when x = 20. This bulk crystallization reduces the glass dissolution without compromising the precipitation of a reactive layer at the glass surface. Finally, in vitro cell tests were performed using MC3T3 pre-osteoblastic cells. While the substitution of CaF2 for CaO led to an increased cytotoxicity, the controlled crystallization of the fluorine containing glasses decreased such cytotoxicity to similar values than traditional bioactive phosphate glass (x0). This study reports on new oxyfluorophosphate glass and glass-ceramics able, not only, to precipitate a Ca-P reactive layer but also to be processed into glass-ceramics with controlled crystal size, density and cellular activity. STATEMENT OF SIGNIFICANCE: Uncontrolled crystallization of bioactive glasses has negative effect on the materials' bioactivity. While in silicate glass the bioactivity is solely reduced, in phosphate glasses it is often completely suppressed. Furthermore, the need for fluorine containing bioactive glasses, not only for use in bone reconstruction but also in toothpaste as emerged. The addition of F in both silicate and phosphate has led to challenges due the lack of Si-F or P-F bonds, generally leading to a decrease in bioactivity. Here, we developed a bioactive invert phosphate glass where up to 20 mol% of CaO was replaced with CaF2. In the new developed glasses, NMR demonstrated formation of P-F bonds. The content of fluorine was tailored to induce CaF2 bulk crystallization. Overall an increase in F was associated with an increase network connectivity. In turns it led to an increased dissolution rate which was linked to a higher cytotoxicity. Upon (partial to full) surface crystallization of the F-free glass, the bioactivity (ability to form a reactive layer) was loss and the cytotoxicity again increased due to the rapid dissolution of one crystal phase and of the remaining amorphous phase. On another hand, the controlled bulk precipitation of CaF2 crystals, in the F-containing glass, was associated with a reduced cytotoxicity. The new oxyfluorophosphate glass-ceramic developed is promising for application in the biomedical field.

Dissolution, bioactivity and osteogenic properties of composites based on polymer and silicate or borosilicate bioactive glass.

Bioactive glass (BAG)/Poly (Lactic Acid) (PLA) composites have great potential for bone tissue engineering. The interest in these materials is to obtain a scaffold with tailorable properties bringing together the advantages of the composites' constituents such as the biodegradability, bioactivity and osteoinduction. The materials studied are PLA/13-93 and PLA/13-93B20 (20% of SiO2 is replaced with B2O3 in the 13-93 composition). To characterize them, they were dissolved in TRIS buffer and Simulated Body Fluid (SBF) in vitro. Over the 10 weeks of immersion in TRIS, the ion release from the composites was constant. Following immersion in SBF for 2 weeks, the hydroxyapatite (HA) layer was found to precipitate at the composites surface. By adding Boron, both these reactions were accelerated, as the borosilicate glass dissolves faster than pure silicate glass alone. Polymer degradation was studied and showed that during immersion, the pure PLA rods maintained their molecular weight whereby the composites decreased with time, but despite this the mechanical properties remained stable for at least 10 weeks. Their ability to induce osteogenic differentiation of myoblastic cells was also demonstrated with cell experiments showing that C2C12 cells were able to proliferate and spread on the composites. The Myosin Heavy Chain and Osteopontin were tracked by immunostaining the cells and showed a suppression of the myosin signal and the presence of osteopontin, when seeded onto the composites. This proves osteoinduction occurred. In studying the mineralization of the cells, it was found that BAG presence conditions the synthesizing of mineral matter in the cells. The results show that these composites have a potential for bone tissue engineering.

Fibrin-polyethylene oxide interpenetrating polymer networks: new self-supported biomaterials combining the properties of both protein gel and synthetic polymer.

Interpenetrating polymer network (IPN) architectures were conceived to improve the mechanical properties of a fibrin gel. Conditions allowing an enzymatic reaction to create one of the two networks in IPN architecture were included in the synthesis pathway. Two IPN series were carried out, starting from two polyethylene oxide (PEO) network precursors leading to different cross-linking densities of the PEO phase. The fibrin concentration varied from 5 to 20 wt.% in each series. The behavior of these materials during dehydration/hydration cycles was also studied. The mechanical properties of the resulting IPN were characterized in the wet and dry states. These self-supported biomaterials combine the properties of both a protein gel and a synthetic polymer. Finally, cells were grown on PEO/fibrin IPN, indicating that they are non-cytotoxic.

Pillaring effects in macroporous carrageenan-silica composite microspheres.

The impregnation of a carrageenan gel by a silica sol is an efficient method to form a composite material which can be conveniently activated by CO2 supercritical drying. The textural properties of the solids have been characterized by nitrogen adsorption-desorption at 77 K and their composition by thermogravimetric analysis and EDX microprobe. Morphology was examined by SEM. The silica-carrageenan composites present an open macroporous structure. Silica particles retained inside the gel behaved as pillars between the polysaccharide fibrils and form a stick-and-ball network. The stiffening of the carrageenan gel by silica prevented its shrinkage upon drying. The nature of the alkali cations affected the retention of silica particles inside the gel. In the absence of silica, carrageenan fibrils rearrange under supercritical drying and form an aerogel with cavities in the mesopore range.

Relationship between upper gastrointestinal symptoms and lesions on CAPD patients.

In order to establish a relationship between upper GI symptoms and CAPD therapy, 34 CAPD patients underwent UGI endoscopy and X-ray studies. Endoscopy is more specific than upper GI X-ray series in the diagnosis of gastrointestinal lesions.