Eric Wers

Physical and biological characteristics of nanohydroxyapatite and bioactive glasses used for bone tissue engineering

Abstract Critical-sized bone defects have, in many cases, posed challenges to the current gold standard treatments. Bioactive glasses are reported to be able to stimulate more bone regeneration than other bioactive ceramics; however, the difficulty in producing porous scaffolds made of bioactive glasses has limited their extensive use in bone regeneration. On the other hand, calcium phosphate ceramics such as synthetic hydroxyapatite and tricalcium phosphate are widely used in the clinic, but they stimulate less bone regeneration. This paper gives an overview of the recent developments in the field of bioactive nanoparticles, with a focus on nanohydroxyapatite and bioactive glasses for bone repair and regeneration. First, a brief overview of the chemical structure and common methods used to produce synthetic nanohydroxyapatite and bioactive glasses has been presented. The main body of the paper covers the physical and biological properties of these biomaterials, as well as their composites with biodegradable polymers used in bone regeneration. A summary of existing challenges and some recommendations for future directions have been brought in the concluding section of this paper.

Excess entropy and thermal behavior of Cu- and Ti-doped bioactive glasses

Bioactive glasses belong to the ceramic family. They are good materials for implantation due to their excellent capacities to create an intimate bond with bones. Copper is known for its anti-inflammatory, antibacterial, and antifungal properties. Titanium is biocompatible and resistant to corrosion. These chemical elements can be introduced in bioactive glasses to provide a wide variety of uses and to enhance the physiological properties of implanted biomaterials. In this work, bioactive glasses doped with different contents of copper and titanium were synthesized by the melting method. The purpose is to study the effect of doping metal element on the thermal characteristics (Tg, Tc, and Tf). The results revealed that the increase of the content of copper and titanium in the glass matrix decreases the melting temperature and induces an increase of the thermal stability. The excess entropies of pure and doped glasses were calculated. Obtained results highlighted the decrease of the excess entropy with the increase of metal elements contents.

Effect of Copper and Zinc on the Bioactivity and Cells Viability of Bioactive Glasses

Bioactive glasses, doped with traces of copper (Cu) and zinc (Zn) were synthesized by fusion method. Cu and Zn present interesting functions for the biological metabolism through their antibacterial, anti-inflammatory and antifungal properties. Several physical methods were employed to characterize the bioactive glasses before and after immersion in a Simulated Body Fluid (SBF). The “in vitro” experiments showed that after soaking in SBF, the behavior of bioactive doped glasses are different compared to pure glass 46S6. Obtained results show that glass matrix undergoes some changes after 15 days of immersion. The non toxic character of doped glasses was confirmed after 24 hours of incubation. The kinetic of release of Cu and Zn was carried out. It highlights that Cu is more released than Zn. By SEM, the morphology of hydroxyapatite obtained with Zn-doped glass show a better crystallization compared to Cu-doped glass.

Chitosan effects on glass matrices evaluated by biomaterial. MAS-NMR and biological investigations

Bioactive glass 46S6 and biodegradable therapeutic polymer (Chitosan: CH) have been elaborated to form 46S6-CH composite by freeze-drying process. The kinetics of chemical reactivity and bioactivity at the surface were investigated by using physicochemical techniques, particularly solid-state MAS-NMR. Immortalized cell line used to construct multicellular spheroids was employed as three-dimensional (3D) cell cultures for in vitro studies. Obtained results showed a novel structure of the composite; the chemical treatment (ultrasound, magnetic stirring, freeze drying process and lyophilization) led the bioactive glass particles to be loaded in the chitosan-based materials. 29Si and 31P MAS-NMR results showed the emergence of two new species, QSi3(OH) and QSi4, which are characteristic of the vitreous network dissolution in simulated body fluid (SBF). MAS-NMR also confirmed the formation of amorphous calcium phosphate (ACP) at the surface of the initial 46S6-CH. Three-dimensional (3D) cell cultures highlighted the effect of chitosan, where the cell viability reached up to 78% in 46S6-CH composite and up to 67% in 46S6. The association of (CH) and bioactive glass (BG) matrix promotes a highly significant bioactivity, demonstrating surface bone formation and satisfactory behavior in biological environment.

Comparative Study of Nanobioactive Glass Quaternary System 46S6

Different bioactive glass systems have been prepared by sol-gel. However, the production of Na2O-containing bioactive glasses by sol–gel methods has proved to be difficult as the sodium nitrate used in the preparation could be lost from the glass structure during filtration and washing. The aim of this study was to prepare the quaternary system 46S6 of bioactive glass by modified sol-gel techniques with a decrease in the time of gelation. In addition, compare the behaviour of the prepared sol-gel bioactive glass system by its corresponding prepared by melting. The obtained glasses were characterized by using several physicochemical techniques; XRD, FTIR, TEM and SEM beside the effect of the glass particles on the viability of osteoblast like cells (Saos-2). Results show that nanopowders 40-60 nm of 46S6 glass system had been prepared by modified sol-gel (acid-base reaction) method at 600°C in just three days at 600°C. Cell viability by MTT assay confirmed the effectiveness of the prepared nanobioactive glass.

New method of synthesis and in vitro studies of a porous biomaterial.

Biomaterials for bone reconstruction represent a widely studied area. In this paper, a new method of synthesis of a porous glass-ceramic obtained by thermal treatment is presented. The prepared biomaterial was characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), and induced couple plasma-optical emission spectroscopy (ICP-OES), mercury porosimetry and by the Archimedes method. In vitro evaluations in a simulated body fluid (SBF) and in contact with SaOS2 human osteoblasts were also carried out. The porous glass-ceramic is composed of a total porous network of 60% suitable for body fluid and cell infiltration, with pore sizes varying from 60 nm to 143 μm. The presence of two crystalline phases decreases the kinetic of bioactivity compared to an amorphous biomaterial (bioactive glass). A hydroxyapatite layer appears from 15 days of immersion on the surface and inside the pores, showing a biodegradation and a bioactivity in four steps. Cytotoxicity assessments present an increase of the cellular viability after 72 h proving the non-cytotoxic effect of the glass-ceramic. Thus, the results of these different studies indicate that the porous biomaterial may have a potential application for the bone regeneration. This paper also presents the novelty of this method. It is a rapid synthesis which combines simplicity and low cost. This represents an advantage for an eventual industrialization.

In vitro chemical and biological effects of Ag, Cu and Cu + Zn adjunction in 46S6 bioactive glasses

Three bioactive glasses belonging to the system SiO2-CaO- Na2O-P2O5 elaborated by conventional melt-quenching techniques were doped with silver, copper and copper + zinc. They were characterized using the usual physical methods. Human osteoblast cells Saos-2 and human endothelial cells EAhy926 were used for viability assays and to assess the metallic ions, self toxicity. Human monocyte cells THP-1 were used to measure interleukins IL1β and IL6 release. Glass chemical structures did not vary much on introduction of metal ions. A layer of hydroxyapatite was observed on every glass after 30 days of SBF immersion. A proliferative action was seen on Saos-2 after 24 h of incubation, EAhy926 growth was not affected. For both cell lines, a moderate cytotoxicity was found after 72 h. Dose-dependent toxic effects of Ag, Cu and Zn ions were observed on Saos-2 and EAhy926 cells. Measured CD50 of silver against these two cell lines were 8 to 20 fold lower than copper and zincs. Except undoped control glass, all doped glasses tested showed anti-inflammatory properties by preventing IL1β and IL6 excretion by differentiated THP-1. In conclusion, strictly monitored adjunction of metal ions to bioglasses ensures good anti-inflammatory properties without altering their biocompatibility.

New hybrid agarose/Cu-Bioglass® biomaterials for antibacterial coatings

Agarose hydrogels, combined with 45S5 Bioglass®, were elaborated to serve as copper delivery systems. Our aim was to study the antibacterial properties of these hydrogels. The results show that the amount of glass does not influence the stiffness properties, but it improves the hydrophilicity and the swelling profile of agarose hydrogel. Two bacterial strains, Bacillus sp. 4J6 and Pseudomonas aeruginosa sp. PAO1, were chosen. Their retention on the substrates was analyzed by confocal laser scanning microscopy. The mechanical characteristics and the release of copper have an effect on the bacterial adhesion and the biofilm formation. All the obtained results indicate that these hydrogels could be adapted to a potential application to the antibacterial coatings