Biocompatibility of glass-crystalline materials obtained by the sol-gel method: effect on macrophage function.

The aim of this work was to confirm in vitro biocompatibility of a new gel-derived glass-crystalline material containing hydroxyapatite and wollastonite phases. For the purpose of comparison, studies were also carried out for a material of the same chemical composition obtained by the traditional melting method. We examined the behaviour and response of cells cultured in the presence of the studied materials. The level of activation of macrophages in culture was determined using three different methods: measurement of respiratory burst by chemiluminescence, nitrite assay and by bioassay of secreted cytokines after immunoelectrophoresis of acute phase proteins from hepatoma cells. All our results show a relatively low, close to control level, activation of macrophages exposed to the studied materials. This indicates a good biocompatibility of both the gel-derived material and the material obtained by the traditional melting method.

Comparison of biocompatibility of gel-derived bioactive ceramics in macrophage culture conditions.

The behaviour of rat alveolar macrophages cultured in the presence of three new gel-derived ceramic biomaterials (CaO-P2O5-SiO2 system) with slightly different chemical compositions was examined. The abilities of these three materials to stimulate alveolar macrophages were compared. Non-treated and lipopolysaccharide-treated macrophages were used as control. The level of macrophage activation was determined by nitrite and prostaglandin E2 assay and respiratory burst measurement by chemiluminescence. The results of these studies showed different macrophage responses to these three relatively similar stimuli. Two of the studied materials were shown to be potent activators of respiratory burst and prostaglandin E2 secretion without any significant release of nitric oxide. On the contrary, the material characterized as the most surface reactive strongly affected only nitric oxide generation by the cells.

Evaluation of biocompatibility of apatite-wollastonite ceramics in fibroblast cultures.

The aim of this work has been to test the biocompatibility of four bioactive, gel derived glass-ceramic materials of CaO-PO2-SiO2 system, modified by addition of boron, aluminum and magnesium compounds. We have examined the growth, collagen synthesis, adhesion and morphology of NRK rat fibroblasts cultured in direct and indirect contact with biomaterials. The growth of cells cultures has been quantified by two methods: [3H]thymidine incorporation and direct counting of cells. The level of collagen synthesis has been used as a parameter describing metabolic activity of cells. Cellular morphology has been assessed following 24 h and 4 days of culturing cells on biomaterials by using SEM and confocal microscopy, respectively. Additionally, in order to obtain information about the attachment of cells to substratum the presence of focal contacts has been examined. The results of all the experiments have demonstrated that none of the materials under study significantly altered cellular functions that were tested. This indicates that additions of MgO, Al2O3 and B2O3 have not induced cytotoxicity of the materials under study. This qualifies them for further in vivo experiments.

Incorporation of sol-gel bioactive glass into PLGA improves mechanical properties and bioactivity of composite scaffolds and results in their osteoinductive properties.

In this study, 3D porous bioactive composite scaffolds were produced and evaluated for their physico-chemical and biological properties. Polymer poly-L-lactide-co-glycolide (PLGA) matrix scaffolds were modified with sol-gel-derived bioactive glasses (SBGs) of CaO-SiO2-P2O5 systems. We hypothesized that SBG incorporation into PLGA matrix would improve the chemical and biological activity of composite materials as well as their mechanical properties. We applied two bioactive glasses, designated as S2 or A2, differing in the content of SiO2 and CaO (i.e. 80 mol% SiO2, 16 mol% CaO for S2 and 40 mol% SiO2, 52 mol% CaO for A2). The composites were characterized for their porosity, bioactivity, microstructure and mechanical properties. The osteoinductive properties of these composites were evaluated in human bone marrow stromal cell (hBMSC) cultures grown in either standard growth medium or treated with recombinant human bone morphogenetic protein-2 (rhBMP-2) or dexamethasone (Dex). After incubation in simulated body fluid, calcium phosphate precipitates formed inside the pores of both A2-PLGA and S2-PLGA scaffolds. The compressive strength of the latter was increased slightly compared to PLGA. Both composites promoted superior hBMSC attachment to the material surface and stimulated the expression of several osteogenic markers in hBMSC compared to cells grown on unmodified PLGA. There were also marked differences in the response of hBMSC to composite scaffolds, depending on chemical compositions of the scaffolds and culture treatments. Compared to silica-rich S2-PLGA, hBMSC grown on calcium-rich A2-PLGA were overall less responsive to rhBMP-2 or Dex and the osteoinductive properties of these A2-PLGA scaffolds seemed partially dependent on their ability to induce BMP signaling in untreated hBMSC. Thus, beyond the ability of currently studied composites to enhance hBMSC osteogenesis, it may become possible to modulate the osteogenic response of hBMSC, depending on the chemistry of SBGs incorporated into polymer matrix.

Behavior of bone marrow cells cultured on three different coatings of gel-derived bioactive glass-ceramics at early stages of cell differentiation.

Previous studies have shown different macrophage responses to three compact pellets (with slightly different chemical composition) of gel-derived bioactive glass-ceramics of the CaO-P2O5-SiO2 system. In the present study primary bone marrow cells directed in vitro to form osteoblastic or osteoclastic cells were cultivated on glass slides coated by these three glass-ceramics. Glass slides were used as controls. In osteoblastic cultures alkaline phosphatase activity varied, depending on the type of coatings. Northern analysis showed high mRNA expressions of bone-related proteins on both the glass-ceramics and control glass. Mineralized nodules were not formed on the control glass, but coating glass slides with the glass-ceramics promoted mineralization without any substantial differences between the types of coatings. In osteoclastic cultures, the normal morphology of tartrate resistant acid phosphatase-positive multinucleated cells on standard culture plates was altered on the control glass and the glass-ceramics. The number of these cells differed, depending on the type of coatings, with no particular differences in the arrangement of F-actin by these cells. These analyses demonstrate complete biocompatibility of the glass-ceramic coatings but not the control glass, on which the cells failed to form mineralized nodules. The phenotype expression of the cells appeared to be influenced by microstructure, surface roughness, and the general character of the coatings rather than their surface reactivity.

Gel-derived materials of a CaO-P(2)O(5)-SiO(2) system modified by boron, sodium, magnesium, aluminum, and fluorine compounds.

Bioactive glass-ceramic materials of the CaO-P(2)O(5)-SiO(2) system modified by adding boron, magnesium, sodium, fluorine, and aluminum were obtained using the sol-gel method. Gel-derived materials were produced in the pellet form obtained by compression of powders as well as in coatings on glass slides. The materials obtained were examined in vitro with regard to the ability of calcium phosphate layer to form on the material surface as the result of contact with simulated body fluid (SBF). SBF pH changes and calcium solubility in this solution were determined and scanning electron microscopy, energy-dispersive X-ray analysis, and infrared spectroscopy studies were conducted before and after contact of the materials with SBF. The gels modified by aluminum were amorphous, whereas the sodium and fluorine additives promoted the bulk crystallization of gel-derived materials. The ability of calcium phosphates to crystallize on the surface of gel-derived materials depended only slightly on the types of additives applied, and the character of this dependence was different from that observed in melted glasses. Moreover, to estimate the biocompatibility of gel-derived coatings, we examined the proliferation, collagen synthesis, adhesion, and morphology of fibroblasts (NRK cells) cultured in the presence of gel-derived materials. The results of these experiments showed that none of the tested materials significantly reduced any cell function.