Elena Landi

Densification behaviour and mechanisms of synthetic hydroxyapatites

Starting from Ca(OH)2 and H3PO4 hydroxyapatite powders with three different crystallinity degrees have been prepared and characterized. Densification extent and mechanisms were studied through dilatometric measurements in isothermal regime in the range of temperature 750–1250°C: the influence of different powder features (including the effect of calcination treatment) have been evaluated. Powder characterized by the lowest crystallinity degree has the highest densification extent; overlapping phenomena occurring during the sintering treatments are responsible for unexpectedly low values found for the shrinkage rate, which can easily lead to a misidentification of the rate controlling mechanism. An interpretation of the densification mechanism, consistent with all experimental findings, is proposed.

Carbonated hydroxyapatite as bone substitute

Abstract B-carbonateapatite (CHA) powder was synthesized starting from calcium nitrate tetrahydrate, diammonium hydrogen phosphate and sodium hydrogen carbonate. The powder was fully characterized in terms of phase purity, stoichiometry, morphology, specific surface area and particle size distribution. The thermal stability of the powder in air and CO 2 atmosphere also was evaluated by thermal analysis. Electroacoustic analysis of the water based suspension of the CHA powder was used to determine the stability of the slurry. Porous bodies of CHA were prepared by impregnation of cellulose sponges with a proper slurry of the powder and optimizing the subsequent sintering. The fired samples were characterized in terms of phase purity and carbonate content, microstructure and pore size distribution. The compressive strength also was evaluated, resulting in 6.0±0.5 MPa. First results of in vivo tests on New Zealand White rabbits showed good biocompatibility and osteointegration of the CHA implant, with higher osteoconductive properties and earlier bioresorption, compared to HA samples, used as control.

A novel route in bone tissue engineering: Magnetic biomimetic scaffolds

In recent years, interest in tissue engineering and its solutions has increased considerably. In particular, scaffolds have become fundamental tools in bone graft substitution and are used in combination with a variety of bio-agents. However, a long-standing problem in the use of these conventional scaffolds lies in the impossibility of re-loading the scaffold with the bio-agents after implantation. This work introduces the magnetic scaffold as a conceptually new solution. The magnetic scaffold is able, via magnetic driving, to attract and take up in vivo growth factors, stem cells or other bio-agents bound to magnetic particles. The authors succeeded in developing a simple and inexpensive technique able to transform standard commercial scaffolds made of hydroxyapatite and collagen in magnetic scaffolds. This innovative process involves dip-coating of the scaffolds in aqueous ferrofluids containing iron oxide nanoparticles coated with various biopolymers. After dip-coating, the nanoparticles are integrated into the structure of the scaffolds, providing the latter with magnetization values as high as 15 emu g(-)(1) at 10 kOe. These values are suitable for generating magnetic gradients, enabling magnetic guiding in the vicinity and inside the scaffold. The magnetic scaffolds do not suffer from any structural damage during the process, maintaining their specific porosity and shape. Moreover, they do not release magnetic particles under a constant flow of simulated body fluids over a period of 8 days. Finally, preliminary studies indicate the ability of the magnetic scaffolds to support adhesion and proliferation of human bone marrow stem cells in vitro. Hence, this new type of scaffold is a valuable candidate for tissue engineering applications, featuring a novel magnetic guiding option.

Design of graded biomimetic osteochondral composite scaffolds

With the ultimate goal to generate suitable materials for the repair of osteochondral defects, in this work we aimed at developing composite osteochondral scaffolds organized in different integrated layers, with features which are biomimetic for articular cartilage and subchondral bone and can differentially support formation of such tissues. A biologically inspired mineralization process was first developed to nucleate Mg-doped hydroxyapatite crystals on type I collagen fibers during their self-assembling. The resulting mineral phase was non-stoichiometric and amorphous, resembling chemico-physical features of newly deposited, natural bone matrix. A graded material was then generated, consisting of (i) a lower layer of the developed biomineralized collagen, corresponding to the subchondral bone, (ii) an upper layer of hyaluronic acid-charged collagen, mimicking the cartilaginous region, and (iii) an intermediate layer of the same nature as the biomineralized collagen, but with a lower extent of mineral, resembling the tidemark. The layers were stacked and freeze-dried to obtain an integrated monolithic composite. Culture of the material for 2 weeks after loading with articular chondrocytes yielded cartilaginous tissue formation selectively in the upper layer. Conversely, ectopic implantation in nude mice of the material after loading with bone marrow stromal cells resulted in bone formation which remained confined within the lower layer. In conclusion, we developed a composite material with cues which are biomimetic of an osteochondral tissue and with the capacity to differentially support cartilage and bone tissue generation. The results warrant testing of the material as a substitute for the repair of osteochondral lesions in orthotopic animal models.

Intrinsic magnetism and hyperthermia in bioactive Fe-doped hydroxyapatite

The use of magnetic activation has been proposed to answer the growing need for assisted bone and vascular remodeling during template/scaffold regeneration. With this in mind, a synthesis procedure was developed to prepare bioactive (Fe2+/Fe3+)-doped hydroxyapatite (Fe-HA), endowed with superparamagnetic-like properties. This new class of magnetic hydroxyapatites can be potentially employed to develop new magnetic ceramic scaffolds with enhanced regenerative properties for bone surgery; in addition, magnetic Fe-HA can find application in anticancer therapies, to replace the widely used magnetic iron oxide nanoparticles, whose long-term cytotoxicity was recently found to reach harmful levels. An extensive physicochemical, microstructural and magnetic characterization was performed on the obtained Fe-HA powders, and demonstrated that the simultaneous addition of Fe2+ and Fe3+ ions during apatite nucleation under controlled synthesis conditions induces intrinsic magnetization in the final product, minimizing the formation of magnetite as secondary phase. This result potentially opens new perspectives for biodevices aimed at bone regeneration and for anti-cancer therapies based on hyperthermia.

Biologically inspired growth of hydroxyapatite nanocrystals inside self-assembled collagen fibers

Abstract Bone defects are generally filled using autologous implants because artificial bone materials have low bioaffinity. However, natural bone can induce infections and antigenic reaction, therefore, the preparation of artificial material with composition, structure and biological feature comparable to those of bone is a goal to be pursued. The aim of this work was to follow a biologically inspired approach performing a direct nucleation of hydroxyapatite (HA) on self-assembled collagen fibers to set up a collagen–hydroxyapatite nanocrystals composite as a new particularly attractive material for bone repair and reconstruction. X-ray diffractometric technique, thermogravimetric (TG–DTG), spectroscopic (FT-IR, ICP), microscopic (SEM, TEM) analyses have been used to highlight the likeness of the artificial biomimetic HA/Col composite with natural bone tissue.

Porous hydroxyapatite/gelatine scaffolds with ice-designed channel-like porosity for biomedical applications

A cryogenic process, including freeze-casting and drying has been performed to obtain hydroxyapatite (HA) scaffolds (approx. diameter 10 mm, height 20 mm) with completely lamellar morphology due to preferentially aligned channel-like pores. Changing the process parameters that influence the cold transmission efficiency from the bottom to the top of the poured HA slurry, lamellar ice crystals with different thickness grew throughout the samples. After sintering, scaffolds with porosity features nearly resembling the ice ones were obtained. The interconnection of pores and the ability of the scaffolds to be rapidly penetrated by synthetic body fluid has been proven. Biohybrid HA/gel composites were prepared, infiltrating HA lamellar scaffolds (45-55 vol.% of porosity) with a 10wt.% solution of gelatine. Colouring genipine was used to cross-link gelatine and clearly show the distribution of the protein in the composite. The compressive mechanical properties of lamellar scaffolds improved with the addition of gelatine: the strength increased up to 5-6 times, while the elastic modulus and strain approximately doubled. The effectiveness of the cross-linkage has been preliminarily verified following scaffold degradation in synthetic body fluid.

Sintering and characterization of HA and TCP bioceramics with control of their strength and phase purity

HA and β-TCP-based ceramics were prepared using commercial powders. Powder characteristics were defined and the processing parameters studied, aimed at the production of samples with improved microstructural and mechanical properties. The behaviour of HA powder subjected to various thermal treatments was investigated in order to control the formation of secondary phases (α- and β-TCP) during sintering. The optimal thermal treatment required to prepare pure β-TCP powder from the precursors (HA and DCP) was determined and the sintering method required to prepare fully dense β-TCP completely free from α-form, was identified. Translucent hot-pressed β-TCP ceramics with potential applications in aesthetic restorative prostheses were prepared and characterized. The interval of existence of α-TCP and α-TCP as secondary products was also defined. Crystallographic analysis was carried out on the imperfectly known low-temperature α-TCP phase, and a proper monoclinic unit cell determined.

A conceptually new type of bio-hybrid scaffold for bone regeneration

Magnetic bio-hybrid porous scaffolds have been synthesized, nucleating nano-apatite in situ on self-assembling collagen, in the presence of magnetite nano-particles. The magnetic phase acted as a sort of cross-linking agent for the collagen, inducing a chemico-physical-mechanical stabilization of the material and allowing us to control the porosity network of the scaffold. Gradients of bio-mineralization and magnetization were also developed for osteochondral application. The good potentiality of the material as a biomedical device, able to offer assistance to bone regeneration through scaffold reloading with specific factors guided by an external magnetic field, has been preliminarily investigated. Up to now the proof of this concept has been realized through in vitro assessments.

Oxidation behavior of aluminum nitride

The evaluation of the thermal stability of three different fully dense AlN materials in the temperature range of 600 °C to 1400 °C in air indicates the strong effect of the starting composition on the oxidation process. The oxidation resistance of pure AlN and Y 2 O 3 -doped AlN was found to be good up to ≍1350 °C. The kinetics are linear (1100 ≤ T ≤ 1400 °C) and the process is governed by a surface reaction that gives rise to the formation of a porous, nonprotective oxide scale, where Al 2 O 3 and Y-aluminates (i.e., AlN–Y 2 O 3 ) have been found as crystalline reaction products. For AlN-CaC 2 , higher oxidation rates indicate that the outward migration of Ca modifies the reaction mechanisms. Linear kinetics in the range 1100 ≤ T ≤ 1200 °C are followed by parabolic kinetics at higher temperatures ( T > 1250 °C); with regard to the latter behavior, an activation energy of 160 kJ/mole could indicate the diffusion of oxygen through the oxidation scale as the rate-controlling mechanism.