Fluoroapatite glass-ceramic coating on alumina: surface behavior with biological fluids

The results of a surface analysis performed on a fluoroapatite-based glass ceramic (SAF) also coating a full-density alpha-alumina substrate (SAF-alumina coating) are presented. These two materials have also been evaluated after soaking in simulated body fluid to understand their ability to induce hydroxyapatite growth on them. Aiming to understand the fluoroapatite glass-ceramic interaction with some plasma proteins, in the second part of this study, fibronectin, albumin, immunoglobulin G, IgA, and complement factor C3c SAF binding have been evaluated; surface activity on complement activation has also been quantified. SAF-alumina coating provides good sites for the nucleation and growth of an apatite layer, equivalent to the mineral component of bone and binds preferentially plasma fibronectin, which is well known to enhance cell adhesion and spreading. Moreover, SAF-alumina coating reduces alumina complement activation directly or via reduced IgA binding. Alumina was shown to bind the same C3 fragments as Zymosan, used as complement activating control, and to induce increased levels of serum soluble iC3b and Bb. A mechanical resistant material with enhanced bioactivity, bone integration, and reduced inflammatory potential respect to alumina has been obtained.     

Bioactive polymethyl methacrylate-based bone cement: comparison of glass beads, apatite- and wollastonite-containing glass-ceramic, and hydroxyapatite fillers on mechanical and biological properties

A new bioactive bone cement (designated GBC) consisting of polymethyl methacrylate (PMMA) as an organic matrix and bioactive glass beads as an inorganic filler has been developed. The bioactive beads, consisting of MgO-CaO-SiO(2)-P(2)O(5)-CaF(2) glass, have been newly designed, and a novel PMMA powder was selected. The purpose of the present study was to compare this new bone cement GBC's mechanical properties in vitro and its osteoconductivity in vivo with cements consisting of the same matrix as GBC and either apatite- and wollastonite-containing glass-ceramic (AW-GC) powder (designated AWC) or sintered hydroxyapatite (HA) powder (HAC). Each filler added to the cements amounted to 70 wt %. The bending strength of GBC was significantly higher than that of AWC and HAC (p < 0.0001). Cements were packed into intramedullar canals of rat tibiae in order to evaluate osteoconductivity as determined by an affinity index. Rats were sacrificed at 2, 4, and 8 weeks after operation. An affinity index, which equaled the length of bone in direct contact with the cement expressed as a percentage of the total length of the cement surface, was calculated for each cement. At each time interval studied, GBC showed a significantly higher affinity index than AWC or HAC up to 8 weeks after implantation (p < 0.03). The value for GBC increased significantly with time up to 8 weeks (p < 0.006). The handling property of GBC was comparable with that of PMMA bone cement. Our study revealed that the higher osteoconductivity of GBC was due to the higher bioactivity of the bioactive glass beads at the cement surface and the lower solubility of the new PMMA powder to MMA monomer. In addition, it was found that the smaller spherical shape and glassy phase of the glass beads gave GBC strong enough mechanical properties to be useful under weight-bearing conditions. GBC shows promise as an alternative with improved properties to the conventionally used PMMA bone cement.     

Development of glass-ceramic scaffolds for bone tissue engineering: characterisation, proliferation of human osteoblasts and nodule formation

Glass-ceramic macroporous scaffolds for tissue engineering have been developed using a polyurethane sponge template and bioactive glass powders. The starting glass (CEL2) belongs to the system SiO(2)-P(2)O(5)-CaO-MgO-Na(2)O-K(2)O and has been synthesised by a conventional melting-quenching route. A slurry of CEL2 powder, polyvinyl alcohol and water has been prepared in order to coat, by impregnation, the polymeric template. An optimised thermal treatment was then use to remove the sponge and to sinter the glass powders, leading to a glass-ceramic replica of the template. Morphological observations, image analyses, mechanical tests and in vitro tests showed that the obtained devices are good candidates as scaffolds for bone-tissue engineering, in terms of pore-size distribution, pore interconnection, surface roughness, and both bioactivity and biocompatibility. In particular, a human osteoblast cell line (MG-63) seeded onto the scaffold after a standardised preconditioning route in simulated body fluid showed a high degree of cell proliferation and a good ability to produce calcium nodules. The obtained results were enhanced by the addition of bone morphogenetic proteins after cell seeding.     

Synchrotron radiation techniques for structural characterisation of biological entities: an example with renal stone analysis

This paper describes the opportunities given by the synchrotron radiation techniques regarding the structural characterisation of biological entities. After a short recall on the characteristics of the synchrotron radiation, are described the experimental devices based on fluorescence X, wide angle X-ray scattering and X-ray absorption spectroscopy, which may applied for biological samples, especially in the field of stone analysis. Recent progresses in medical research using synchrotron radiation will be also discussed.     

The effect of different powder particle size on mechanical properties of sintered alumina, resin- and glass-infused alumina

In this study, the compaction and sintering behavior of fine alumina powders of different particle sizes and the effect of matrix particle size on biaxial strength and fracture toughness of infused matrices were investigated. Three different alumina powders, In-Ceram alumina, A16SG, and RC172 were selected, representing a range of particle size and shape. RC172 and A16SG were dry-pressed. In-Ceram alumina was slip-cast following manufacturer's recommendations. Dry-pressed ceramic blocks were sectioned into disks with a thickness of 1.5-mm. Uninfused disks were sintered at four temperatures between 1250 degrees C and 1400 degrees C. For glass or resin infused specimens, alumina disks were sintered at 1250 degrees C for 2 h and separated into two groups for glass infusion and resin (UDMA/TEGDMA) infusion. Disks were tested for biaxial flexural strength with a universal testing machine (Instron) at 0.5-mm/min crosshead speeds. One-way ANOVA and Duncan's multiple range tests revealed that alumina disks with different smaller particle sizes have significantly higher biaxial strength (p < 0.05). The strength of the alumina matrix was greatly increased by glass and resin infusion. The biaxial strength of resin-infused alumina increased as particle size decreased, whereas strength of glass-infused alumina was constant.     

Composites consisting of poly(methyl methacrylate) and alumina powder: an evaluation of their mechanical and biological properties

We developed new composites consisting of comparatively high molecular weight poly(methyl methacrylate) (hPMMA) and delta-alumina powder or alpha-alumina powder (designated delta-APC and alpha-APC, respectively) that allowed direct bone formation on their surfaces in vivo. delta-Alumina powder was manufactured by the fusing and quenching of pulverized alumina powder. It was composed mainly of delta-crystal phases of alumina. The purpose of this study was to evaluate the static mechanical properties and biological properties of these composites. The hPMMA itself was used as a reference material. The bending strength and Young's modulus of both delta-APC and alpha-APC were significantly higher than those of hPMMA, and the alumina composites are believed to be strong enough for use under weight-bearing conditions. The three types of composites were packed into the intramedullary canals of rat tibiae to evaluate osteoconductivity, as determined by an affinity index. Rats were sacrificed 4 and 8 weeks after surgery. The affinity index, equal to the length of bone in direct contact with the composite surface and expressed as a percentage of the total length of the composite surface, was calculated for each composite at each interval. Histologically, new bone had formed along the surfaces of both delta-APC and alpha-APC within 4 weeks. The affinity indices for both delta-APC and alpha-APC increased significantly with time up to 8 weeks. At 8 weeks, the affinity index for delta-APC was significantly higher than the indices for alpha-APC and hPMMA. This study revealed that the excellent osteoconductivity of delta-APC was due to the delta-crystal phases of alumina and the high molecular weight of hPMMA. delta-APC shows promise as a base for developing a highly osteoconductive and mechanically strong biomaterial.     

The structural characteristics and mechanical behaviors of nonstoichiometric apatite coatings sintered in air atmosphere.

Two types of nonstoichiometric apatite coatings on Ti6Al4V substrates were prepared in our laboratory by electrodeposition and hydrothermal synthesis. One was composed of highly pure needle-like calcium-deficient hydroxyapatite, and the other consisted of needle-like calcium-deficient hydroxyapatite and plate-like brushite. In this paper, the morphology, phase transition, and bonding strength of these coatings sintered at 450( degrees ) approximately 980 degrees C in an air atmosphere were studied. In addition, the residual stresses in pure calcium-deficient hydroxyapatite coatings and coatings sintered at 800 degrees C were measured by a hole-drilling method. The results show that after being sintered at a higher temperature, the coating/Ti6Al4V samples exhibited structural characteristics such as biphasic [hydroxyapatite + beta-Ca3(PO4)2] coatings and dense TiO2 film/Ti6Al4V in which the residual stresses in the coatings slightly increased. In addition, the results indicate that bonding strength was improved.     

Sol-gel derived aluminosilicate coatings on alumina as substrate for osteoblasts

Rat bone marrow stromal cell differentiation on aluminosilicate 3Al₂O₃–2SiO₂ coatings was investigated. Thin ceramic coatings were prepared on -alumina substrates by the sol–gel process and calcined in order to establish an amorphous aluminosilicate ceramic phase with and without nanosized transitional mullite crystals. In addition, coatings of thermally sprayed aluminosilicate and diphasic γ-alumina–silica nanosized colloids were prepared. Cell culture testing by rat osteoblasts showed good biocompatibility for aluminosilicates with sustained normal osteoblast functions. Despite mutual disparities in physical and chemical nanostructures, the culture findings suggested fairly similar osteoblast response to all tested coatings. The results suggest that topographical frequency parameters and chemical uniformity are important parameters in determining the best conditions for osteoblasts on sol–gel derived aluminosilicate materials.     

Effect of heat pressing on the mechanical properties of a mica-based glass-ceramic

Previous work has shown that heat pressing of mica-based glass-ceramics can lead to crystal alignment along the direction of pressing. The purpose of this study was to evaluate the effect of heat pressing on the fracture toughness of mica-based glass-ceramics. Glass rods (12 x 60 mm) were prepared by melting the glass composition at 1400 degrees C for 2 h. Ingots (12 x 12 mm; n = 5) and discs (12 x 1.5 mm; n = 10) were cut from the rods. The discs were heat treated to simulate heat pressing and served as controls. A machinable mica-based glass- ceramic (Dicor MGC) also served as control. Bar-shaped wax patterns (2 x 4 x 22 mm) were invested and heat pressed at 875 degrees C. The elastic constants were determined with the use of the pulsed ultrasonic velocity method. The fracture toughness was measured by the indentation strength technique. The degree of texture was assessed qualitatively by X-ray diffraction and quantitatively on digital SEM micrographs. The results showed that the mean fracture toughness of the heat-pressed specimens (1.96 +/- 0.19 MPa. m(0.5)) was significantly higher than that of the heat-treated controls (1.51 +/- 0.21 MPa. m(0.5)) or the proprietary mica glass-ceramic (Dicor MGC; 1.66 +/- 0.04 MPa. m(0.5)) (p <.001). It was concluded that heat pressing led to a significant increase in fracture toughness in mica glass-ceramics because of crystal alignment along the direction of pressing.     

Hydroxyapatite and titania sol-gel composite coatings on titanium for hard tissue implants; mechanical and in vitro biological performance

Hydroxyapatite (HA) composites with titania (TiO2) up to 30 mol % were coated on a titanium (Ti) substrate by a sol-gel route, and the mechanical and biological properties of the coating systems were evaluated. Using polymeric precursors, highly stable HA and TiO2 sols were prepared prior to making composite sols and coatings. Coatings were produced under a controlled spinning and heat treatment process. Pure phases of HA and TiO2 were well developed on the composites after heat treatment above 450 degrees C. The HA-TiO2 composite coating layers were homogeneous and highly dense with a thickness of about 800-900 nm. The adhesion strength of the coating layers with respect to Ti substrate increased with increasing the TiO2 addition. The highest strength obtained was as high as 56 MPa, with an improvement of about 50% when compared to pure HA (37 MPa). The osteoblast-like cells grew and spread actively on all the composite coatings. The proliferation and alkaline phosphatase (ALP) activity of the cells grown on the composite coatings were much higher than those on bare Ti, and even comparable to those on pure HA coating. Notably, the HA-20% TiO2 composite coating showed a significantly higher proliferation and ALP expression compared to bare Ti (p < 0.05). These findings suggest that the sol-gel-derived HA-TiO2 composite coatings possess excellent properties for hard tissue applications from the mechanical and biological perspective.     

Functional, molecular and structural characterisation of five anti-Brucella LPS mAb

The O-antigen of the gram negative bacteria Brucella is composed of an homopolymer of 4,6-dideoxy-4-formamido-alpha-D-mannopyranosyl (or perosamine). Several mAb interact specifically with only the O-antigen of certain Brucella species. Although, many studies show that this specific recognition results mainly from the ratios of alpha 1-2 and alpha 1-3 link between the different Brucella strain perosamine residues, little is known about the mAb recognising this O-antigen. In this paper, we describe the binding profile of five anti-Brucella O-antigen mAb to the LPS of two Brucella strains and a bacteria possessing a nearly identical O-antigen: Yersinia enterocolitica 0:9. We show that the specificity of these five mAb can be correlated to their germ line gene usage. Besides, their relative affinity to the different LPS is correlated to their ability to protect against Brucella infection by passive transfer in a mouse model. The analysis of their 3D structure gives new hypothesis of the epitopes recognised.     

DLC coatings: effects of physical and chemical properties on biological response

Recent trials on diamond-like carbon (DLC) coated medical devices have indicated promise for blood interfacing applications. The literature is sparse regarding structural and compositional effects of DLC on cellular response. An important goal in optimizing blood-interfacing implants is minimal macrophage attachment, and maximal albumin:fibrinogen adsorption ratio. DLC coatings deposited by PACVD and FAD, were analysed with respect to sp3 content (EELS), hydrogen content (ERDA), surface composition (XPS), surface roughness (AFM), surface energy, albumin:fibrinogen adsorption ratio, and macrophage viability and attachment. We found that increasing surface roughness and surface energy enhanced the macrophage viability and the albumin:fibrinogen adsorption ratio. We also found that the higher the hydrogen content for a-C:Hs deposited by PACVD, the lower the albumin:fibrinogen adsorption ratio, and macrophage attachment. This suggests that hydrogen content may be an important factor for influencing the biological response of DLC surfaces. Macrophage cells spread well on all DLC surfaces, and the surface results indicated the non-toxic nature of the surfaces on the cells at the time points tested.     

Inhibition of Ni release from NiTi alloy by hydroxyapatite,alumina, and titanium sputtered coatings

To alleviate the effects of Ni allergy from NiTi alloy implants, hydroxyapatite (Ca10(PO4)6(OH)2; HA), alumina (Al2O3), or titanium (Ti) was coated onto NiTi alloy plates to form 1-microm thick films using radio frequency magnetron sputtering. The coatings on the plates were characterized using XRD. After the plates had been immersed in physiological saline for periods of one, four, or eight weeks, the concentration of Ni ions released in each solution was detected using a microwave induced plasma mass spectrometer. After eight weeks, the concentration of Ni ions released from the non-coated, the Ti-coated, the HA-coated, and the alumina-coated plates were 238, 19.7, 183, and 106 ppb, respectively. The bonding strength of the Ti film, the HA film, and the alumina film to the NiTi substrate were 3.8 +/- 1.2, 2.6 +/- 0.7, and 3.1 +/- 1.2 MPa, respectively.The non-coated, the HA-coated, the alumina-coated, and the Ti-coated plates were implanted into the femurs of a dog for four weeks for histological observation. In case of the non-coated plates, connective tissue more than 300 microm thick was observed, whereas for the coated plates the thickness of the connective tissue was around 100 microm.     

Composite lipid polyelectrolyte capsules templated on red blood cells: fabrication and structural characterisation

Charged phospholipids and mixtures of charged phospholipids with zwitterionic lipids were adsorbed onto polyelectrolyte capsules templated on erythrocytes. The assembly was proved by means of electrophoretic mobility measurements, confocal laser scanning microscopy and flow cytometry. Freezefracture electron microscopy proved that the phospholipids assemble as bilayers or multilayers. Single particle light scattering showed that bilayers composed of anionic lipids can be intercalated between subsequent polyelectrolyte inter-layers in a regular manner. Neutral lipids can form multilayers. A pronounced decrease in capsule permeability for small polar dyes upon lipid adsorption was followed by confocal laser scanning microscopy.     

The effects of collagen concentration and crosslink density on the biological,structural and mechanical properties of collagen-GAG scaffolds for bone tissue engineering

In this study, we examined the effects of varying collagen concentration and crosslink density on the biological, structural and mechanical properties of collagen-GAG scaffolds for bone tissue engineering. Three different collagen contents (0.25%, 0.5% and 1% collagen) and two different dehydrothermal (DHT) crosslinking processes [1] 105 ^°C for 24 h and [2] 150 ^°C for 48 h were investigated. These scaffolds were assessed for (1) pore size, (2) permeability (3) compressive strength and (4) cell viability. The largest pore size, permeability rate, compressive modulus, cell number and cell metabolic activity was all found to occur on the 1% collagen scaffold due to its increased collagen composition and the DHT treatment at 150 ^°C was found to significantly improve the mechanical properties and not to affect cellular number or metabolic activity. These results indicate that doubling the collagen content to 1% and dehydrothermally crosslinking the scaffold at 150 ^°C for 48 h has enhanced mechanical and biological properties of the scaffold making it highly attractive for use in bone tissue engineering.     

Calcium phosphate invert glass-ceramic coatings joined by self-development of compositionally gradient layers on a titanium alloy

A glass-ceramic layer containing beta-Ca3(PO4)2 crystals could be joined easily with a new type of titanium alloy (Ti-29Nb-13Ta-4.6Zr) consisting of a beta-titanium phase by heating the metal, on which glass powders with a composition of 60CaO x 30P2O5 7Na2O x 3TiO2 were placed, at 800 degrees C in air. Measurement of tensile bonding strength revealed that the joining between the coating layer and the substrate is very strong. Even after the large deformation (e.g., approximately 90 degrees in bending angle) of the titanium alloy, the coating layer was not peeled off from the substrate. A compositionally gradient layer in the TiO2-P2O5-Na2O-CaO system is developed automatically on the titanium alloy during the heating, resulting in the formation of the strong joining. By soaking in simulated body fluid at 37 degrees C, hydroxyapatite phase was formed newly on the surface of the coating layer.     

A new reconstituted connective tissue matrix: preparation, biochemical, structural and mechanical studies

A fibrinogen derivative generated by thrombin was reacted with elastin to yield a new addition product or adduct between the two proteins. Addition of fibronectin, and then of collagen, did not interfere with the basic elastin-fibrinogen reaction and conferred the qualities of an artificial connective tissue to the product. Biochemical, structural and biomechanical aspects of the new matrix were studied. Aprotinin, heparin, thiomersal, and thiourea did not inhibit the main reaction; indeed, some of these ingredients improved the matrix cohesion. Scanning electron microscopy showed the genesis of a true network whose meshes were more reticulated by the addition of thiourea. Biomechanical studies, i.e., strength and elasticity showed the thiourea matrix to be the strongest. These intrinsic properties suggest the product could have biological and clinical applications.     

Strengthening of bone-implant interface by the use of granule coatings on alumina ceramics

Three different granule coatings (a granular alumina ceramic coating, a granular hydroxyapatite coating, and a polished granular hydroxyapatite coating) applied to alumina ceramic substrate were evaluated for their strengthening effects of the bone-implant interface in rabbit tibiae. For a comparison, noncoated alumina ceramics, and dense hydroxyapatite were assessed in the same way. The granular alumina ceramic coating, creating a bioinert, porous surface, was most effective due to a strong mechanical bond between the bone and implant. The interface strength was even higher than that of the dense hydroxyapatite. The granular hydroxyapatite coating, creating a bioactive, porous surface, was less effective than the granular alumina ceramic coating because of the brittleness of the hydroxyapatite granules, although it formed a direct and mechanical bond with bone tissue. The polished granular hydroxyapatite coating, creating a bioactive, smooth surface, was least effective because of the brittleness of the hydroxyapatite granules, though it presented an improved interface strength compared with that of the noncoated alumina ceramics due to a direct bond between the bone and hydroxyapatite granules.     

Fabrication, characterization, and biological assessment of multilayer DNA coatings on sandblasted-dual acid etched titanium surface

As local gene therapy has received attention, immobilizing functional gene onto irregular oral implant surface has become an advanced challenge. Electrostatic layer-by-layer (LBL) assembly technique could achieve this goal and allow local and efficient administration of genes to the target cells. In this study, multilayers of cationic lipid/plasmid DNA (pEGFP-C1) complex (LDc) and anionic hyaluronic acid were assembled onto sandblasted-dual acid etched titanium disks by the LBL technique. Surface characteristics of the coatings were performed by x-ray photospectroscopy (XPS), contact angle measurements, and scanning electron microscopy (SEM). The cell biological characteristics of the coatings were evaluated by in vitro experiments. SEM results demonstrated that the porous titanium surface was gradually flattened with the increase of the multilayer. The XPS survey indicated that the N element was found from the coating. The coating degradation and pEGFP-C1 releasing kinetics showed that the more assembled layer numbers were, the larger the amount of DNA released in the first 30 h. MC3T3-E1 cells were cultured directly on the DNA-loaded surface. Higher enhanced green fluorescent protein (EGFP) expression efficiency was achieved by increasing the number of layers when cells were cultured after 24 or 72 h. The MC3T3-E1 cell viability on the surface of multilayer DNA coatings was significantly higher than that on control porous titanium surface. It was concluded that the approach established by the LBL technique had great potential in immobilizing gene coatings onto the porous titanium surface and subsequently influenced the function of the cultured cell.