An experimental and theoretical approach on stability towards hydrolysis of triethyl phosphate and its effects on the microstructure of sol-gel-derived bioactive silicate glass.

The sol-gel method is versatile and one of the well-established synthetic approaches for preparing bioactive glass with improved microstructure. In a successful approach, alkoxide precursors undergo rapid hydrolysis, followed by immediate condensation leading to the formation of three-dimensional gels. On the other hand, a slow kinetics rate for hydrolysis of one or more alkoxide precursors generates a mismatch in the progression of the consecutive reactions of the sol-gel process, which makes it difficult to form homogeneous multicomponent glass products. The amorphous phase separation (APS) into the gel is thermodynamically unstable and tends to transform into a crystalline form during the calcination step of xerogel. In the present study, we report a combined experimental and theoretical method to investigate the stability towards hydrolysis of triethyl phosphate (TEP) and its effects on the mechanism leading to phase separation in 58S bioactive glass obtained via sol-gel route. A multitechnical approach for the experimental characterization combined with calculations of functional density theory (DFT) suggest that TEP should not undergo hydrolysis by water under acidic conditions during the formation of the sol or even in the gel phase. The activation energy barrier (ΔG‡) showed a height of about 20 kcal·mol-1 for the three stages of hydrolysis and the reaction rates calculated for each stage of TEP hydrolysis were kFHR = 7.0 × 10-3s-1, kSHR = 6.8 × 10-3s-1 and kTHR = 3.5 × 10-3s-1. These results show that TEP remains in the non-hydrolyzed form segregated within the xerogel matrix until its thermal decomposition in the calcination step, when P species preferentially associate with calcium ions (labile species) and other phosphate groups present nearby, forming crystalline domains of calcium pyrophosphates permeated by the silica-rich glass matrix. Together, our data expand the knowledge about the synthesis by the sol-gel method of bioactive glass and establishes a mechanism that explains the role played by the precursor source of phosphorus (TEP) in the phase separation, an event commonly observed for these biomaterials.

Could petroleum work as lubricant oil on slippery lubricated surfaces to prevent inorganic scaling?

The use of nucleation and growth inhibitors at offshore oil industry to avoid inorganic scaling could be replaced by both physical and chemical modifications at surfaces to prevent the scaling. In that way, the slippery lubricated surfaces have been showing promising results as scaling preventers, notably when the lubricants are perfluorinated oils, which are immiscible with almost every kind of compound. Nonetheless, the requirement of periodically refilling these lubricant oils is disadvantageous, especially when dealing with deep sea facilities. Using petroleum as the lubricant oil could skip the refilling step, since it is always present in the extraction medium. So, this work tests the effectiveness of petroleum as the lubricant oil in functionalized textured polyaniline applied as anti-scaling material in conditions that simulate the medium of offshore operation. As result, petroleum as lubricant oil presents effective anti-scaling capacity and works perfectly in slippery lubricated surfaces.

Investigation of citric acid-assisted sol-gel synthesis coupled to the self-propagating combustion method for preparing bioactive glass with high structural homogeneity.

In this research, the mechanism of an efficient strategy for the synthesis of 58S bioglass with high structural homogeneity by a citric acid assisted sol-gel route was investigated. This is an interesting approach to prepare bioactive glass via the sol-gel method with application potential in bone tissue engineering and also for the development of new biomedical devices. Herein, 58S bioglass was synthesized by two routes: conventional sol-gel method (CSG) and citric acid assisted sol-gel route coupled to the self-propagating combustion method (SPC). The effects of citric acid on the temperature required for 58S vitreous consolidation, long- and short-range ordering were investigated by several analysis techniques. Results suggested that citric acid molecules serve as an effective molecular template formed by molecular network raised from intermolecular forces, especially the hydrogen bonds, resulting from the chemical interactions between the COOH and hydroxyl groups (water, ethanol, POH, SiOH). In this scenario, citric acid controls the phase segregation during the drying and combustion steps of the gel in the SPC method by establishing chemical interactions (hydrogen bonds) with the superficial silanol groups present on the small-sized silica nanoparticles present in the sol governing their growth. Besides these mentioned features, the self-propagating combustion behavior exhibited by the nitrate-citrate in the SPC xerogel during the combustion step allowed the removal of the organic load and the consolidation of the vitreous structure at a temperature considerably lower than the sample obtained by the CSG method. Consequently, the SPC method leads to the formation of a glass structure with high homogeneity for the 58S, whereas the conventional sol-gel method produces a matrix enriched with calcium phosphate crystalline nuclei - glass-ceramic.

Comprehensive in vitro and in vivo studies of novel melt-derived Nb-substituted 45S5 bioglass reveal its enhanced bioactive properties for bone healing.

The present work presents and discusses the results of a comprehensive study on the bioactive properties of Nb-substituted silicate glass derived from 45S5 bioglass. In vitro and in vivo experiments were performed. We undertook three different types of in vitro analyses: (i) investigation of the kinetics of chemical reactivity and the bioactivity of Nb-substituted glass in simulated body fluid (SBF) by 31P MASNMR spectroscopy, (ii) determination of ionic leaching profiles in buffered solution by inductively coupled plasma optical emission spectrometry (ICP-OES), and (iii) assessment of the compatibility and osteogenic differentiation of human embryonic stem cells (hESCs) treated with dissolution products of different compositions of Nb-substituted glass. The results revealed that Nb-substituted glass is not toxic to hESCs. Moreover, adding up to 1.3 mol% of Nb2O5 to 45S5 bioglass significantly enhanced its osteogenic capacity. For the in vivo experiments, trial glass rods were implanted into circular defects in rat tibia in order to evaluate their biocompatibility and bioactivity. Results showed all Nb-containing glass was biocompatible and that the addition of 1.3 mol% of Nb2O5, replacing phosphorous, increases the osteostimulation of bioglass. Therefore, these results support the assertion that Nb-substituted glass is suitable for biomedical applications.

Addition of Wollastonite Fibers to Calcium Phosphate Cement Increases Cell Viability and Stimulates Differentiation of Osteoblast-Like Cells.

Calcium phosphate cement (CPC) that is based on α-tricalcium phosphate (α-TCP) is considered desirable for bone tissue engineering because of its relatively rapid degradation properties. However, such cement is relatively weak, restricting its use to areas of low mechanical stress. Wollastonite fibers (WF) have been used to improve the mechanical strength of biomaterials. However, the biological properties of WF remain poorly understood. Here, we tested the response of osteoblast-like cells to being cultured on CPC reinforced with 5% of WF (CPC-WF). We found that both types of cement studied achieved an ion balance for calcium and phosphate after 3 days of immersion in culture medium and this allowed subsequent long-term cell culture. CPC-WF increased cell viability and stimulated cell differentiation, compared to nonreinforced CPC. We hypothesize that late silicon release by CPC-WF induces increased cell proliferation and differentiation. Based on our findings, we propose that CPC-WF is a promising material for bone tissue engineering applications.

Facile and innovative method for bioglass surface modification: Optimization studies.

In this work it is presented a facile and novel method for modification of bioglass surface based on (Camolten salt bath2+|Naglass+) ion exchange by immersion in molten salt bath. This method allows changing selectively the chemical composition of a surface layer of glass, creating a new and more reactive bioglass in a shell that surrounds the unchanged bulk of the original BG45S5 bioglass (core-shell type system). The modified bioglass conserves the non-crystalline structure of BG45S5 bioglass and presents a significant increase of surface reactivity in comparison with BG45S5. Melt-derived bioactive glasses BG45S5 with the nominal composition of 46.1mol% SiO2, 24.4mol% Na2O, 26.9mol% CaO, and 2.6mol% P2O5 have been subjected to ion exchange at 480°C in molten mixture of Ca(NO3)2 and NaNO3 with molar ratio of 70:30 for different time periods ranging from 0 to 60min. The optimization studies by using XRF and XRD showed that ion exchange time of 30min is enough to achieve higher changes on the glass surface without alters its non-crystalline structure. The chemical composition, morphology and structure of BG45S5 and bioglass with modified surface were studied by using several analytical techniques. FTIR and O1s XPS results showed that the modification of glass surface favors the formation of Si-ONBO groups at the expense of SiOBOSi bonds. 29Si MAS-NMR studies showed that the connectivity of SiQn species decreases from cross-linked SiQ3 units to chain-like SiQ2 units and finally to depolymerized SiQ1 and SiQ0 units after ion exchange. This result is consistent with the chemical model based on the enrichment with calcium ions of the bioglass surface such that the excess of positive charges is balanced by depolymerization of silicate network. The pH changes in the early steps of reaction of bioactive glasses BG45S5 and BG45Ca30, in deionized water or solutions buffered with HEPES were investigated. BG45Ca30 bioactive glass exhibited a significant increase in the pH during the early steps of the reaction compared to BG45S5.

Hierarchical structures of ß-TCP/45S5 bioglass hybrid scaffolds prepared by gelcasting.

This paper investigates the microstructure and the mechanical properties of ß-tricalcium phosphate (ß-TCP) three-dimensional (3D) porous materials reinforced with 45S5 bioactive glass (BG). ß-TCP and ß-TCP/x%-BG scaffolds with interconnected pores networks, suitable for bone regeneration, were fabricated by gel-casting method. Mechanical properties, porosity, and morphological characteristics were evaluated by compressive strength test, scanning electron microscopy (SEM) and X-ray microtomography analysis, whereas the structures were fully explored by XRD, and Raman spectroscopy. To the best of our knowledge, this is the first time where the mechanism for understanding the effect of bioglass on the mechanical properties and microstruture of ß-TCP/45S5-BG scaffolds has been systematically studied. The findings showed that ionic product lixiviated from 45S5 bioactive glass, rich in silicon species and sodium ion, catalyzes a phase transition from ß-TCP to Si-TCP by replacement of phosphorus for silicon and contributes to the improvement of scaffolds mechanical properties. The compressive strength of ß-TCP/5%-BG and ß-TCP/7.5%-BG was improved around 200% in comparison to pure ß-TCP. Osteoblast-like cells (MG 63) were exposed to the materials for 24h through the use of medium conditioned by ß-tricalcium phosphate/bioactive glass. Cell viability was measured by MTT assay in the cells and the data obtained were submitted to ANOVA, Tukey׳s multiple comparison (p<0.05). The ß-TCP/7.5-BG promoted an increase of cell proliferation. The results suggest that compositions and processing method studied may provide appropriate materials for tissue engineering.

Mechanical, biochemical and morphometric alterations in the femur of mdx mice.

The bone tissue abnormalities observed in patients with Duchenne muscular dystrophy are frequently attributed to muscle weakness. In this condition, bones receive fewer mechanical stimuli, compromising the process of bone modeling. In the present study we hypothesize that other factors inherent to the disease might be associated with bone tissue impairment, irrespective of the presence of muscle impairment. Mdx mice lack dystrophin and present cycles of muscle degeneration/regeneration that become more intense in the third week of life. As observed in humans with muscular dystrophy, bone tissue abnormalities were found in mdx mice during more intense muscle degeneration due to age. Under these circumstances, muscle deficit is probably one of the factors promoting these changes. To test our hypothesis, we investigated the changes that occur in the femur of mdx mice at 21 days of age when muscle damage is still not significant. The mechanical (structural and material) and biochemical properties and morphometric characteristics of the femur of mdx and control animals were evaluated. The results demonstrated a lower strength, stiffness and energy absorption capacity in mdx femurs. Higher values for structural (load and stiffness) and material (stress, elastic modulus and toughness) properties were observed in the control group. Mdx femurs were shorter and were characterized by a smaller cortical area and thickness and a smaller area of epiphyseal trabecular bone. The hydroxyproline content was similar in the two groups, but there was a significant difference in the Ca/P ratios. Thermogravimetry showed a higher mineral matrix content in cortical bone of control animals. In conclusion, femurs of mdx mice presented impaired mechanical and biochemical properties as well as changes in collagen organization in the extracellular matrix. Thus, mdx mice developed femoral osteopenia even in the absence of significant muscle fiber degeneration. This weakness of the mdx femur is probably due to genetic factors that are directly or indirectly related to dystrophin deficiency.

Repair of cranial bone defects with calcium phosphate ceramic implant or autogenous bone graft.

Autogenous bone grafts have frequently been used in the treatment of bone defects; however, this procedure can cause clinical complications after surgery. Besides, the amount of available bone is sometimes insufficient. Therefore, synthetic biomaterials have been researched as an alternative to autogenous bone graft implants. The objective of this study was to evaluate the repair of bone defects treated with compact autogenous bone graft or porous calcium phosphate ceramics. Three defects 3 mm in diameter were produced in the skull of 21 rats. One the defects was produced in the frontal bone, which remained empty, while the others were produced in the right and left parietal bones, which were filled respectively with ceramics and autogenous bone graft. The animals were sacrificed 1, 2, 4, and 24 weeks after surgery and analyzed by light microscopy and radiography. In the twenty-fourth week, the defects filled with autogenous bone graft and ceramics had similar volumes of newly formed bone tissue. The ceramics offered favorable conditions to bone tissue growth. Thus, we concluded that the calcium phosphate ceramic implant proved to be effective in repairing defects produced in the skull of rats.

Subperiosteal hydroxyapatite implants in rats submitted to ethanol ingestion.

It has been reported that excessive alcohol consumption can contribute to failures on the osteointegration process in the site of implantation. Hydroxyapatite blocks were implanted under the periosteum of the femur and skull of 40 rats divided into two groups of 20 animals, one of them received 25% ethanol diluted in water and the other did not. Bone formation close to the hydroxyapatite implant was observed in the femur of all animals 2 weeks after surgery, however the bone volume was lower in ethanol-treated animals. It was observed in the skulls of the ethanol-treated animals a delay in new bone formation process, as a lower bone volume, too. After 4 weeks of the implantation, just one ethanol-treated animal showed no new bone formation in the femur, while no bone formation was observed in the skulls of two other rats. On the 8th and 16th weeks, bone formation was observed in both femur and skull from both groups, although always with less volume in ethanol-treated rats. We concluded that ethanol consumption did not impair osteointegration of ceramic implants, but it might have reduced the osteogenic capacity of periosteal cells in the femur and parietal bone of the rats.