Effect of nitrogen on the antibacterial behavior of oxynitride glasses

The main drawback of bioglasses is their restricted use in load bearing applications and the consequent need to develop stronger glassy materials. This has led to the consideration of oxynitride glasses for numerous biomedical applications. This paper investigated two different types of glasses at a constant cationic ratio, with and without nitrogen (a N containing and a N-free glass composition) to better understand the effect of N on the biological properties of glasses. The results revealed that the addition of N increased the glass transition temperature, isoelectric point (IEP) and slightly increased wettability. Moreover, compared to N including glass, N-free glass exhibited better anti-bacterial activity against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus), two key bacteria that infect implants. In summary, these in vitro results indicated that amine functional groups existing in N containing glasses which are missing in N-free glasses, caused a slight difference in wetting behavior and a more obvious change in isoelectric point and in bacterial response. N-free glasses exhibited better inhibitory results both against E. coli and S. aureus compared to N including glass suggesting that oxygen rich glasses should be further studied for their novel antibacterial properties.     

Hydroxyapatite cages with aligned pores for bone grafting – Seeding of human osteoblast-like cells in vitro and their response in dynamic culture mode

Hydroxyapatite (HA) is a highly regarded synthetic bone graft material. Porous HA ceramics blocks are used to substitute harvested natural bone grafts. Being similar to bone mineral, HA material integrates with the host bone through surface osteointegration and slowly resorb along with the natural bone remodeling process. The blocks in use currently have random and tortuous pore structures. The present work explores the usefulness of cage-like HA ceramic design with end-to-end open pores, with the help of in vitro cell culture methods. Such a structure, on implantation, will take up the blood factors and cells and host the bone remodeling process inside the bulk of the cage, leading to early healing. In the study, HA samples with aligned through-pores were prepared and explored in vitro, with a focus on how the pores host the cells inside and to what level the cells maintain their activity. Human osteoblast-like cells (HOS) were used, at different seeding and culturing approaches. Cell seeding was done through (i) conventional large volume cell suspension, (ii) a confined mini chamber with a limited volume of cell suspension, and (iii) placing a concentrated drop of cell suspension directly on top of the scaffold. The third approach gave the best cell adhesion and proliferation, and hence used for further explorations. A dynamic culture system was designed in-house by bifurcating the cell culture wells using vertical inserts, holding the samples horizontally with their ends open to both sides, and making the media flow across using a rocker platform. The HOS cell adhesion, viability and proliferation were tested in the HA cages, in static and dynamic culture conditions, with conventional porous ceramics as the control. The cell infiltration was deeper and the cell viability over a period of 7 days was significantly higher in dynamic culture conditions in the test samples.     

Synthesis and characterization of biocompatible hydroxyapatite coated ferrite

Ferrite particles coated with biocompatible phases can be used for hyperthermia treatment of cancer. We have synthesized substituted calcium hexaferrite, which is not stable on its own but is stabilized with small substitution of La. Hexaferrite of chemical composition (CaO)_0.75(La₂0₃)_0.20(Fe₂O₃)₆ was prepared using citrate gel method. Hydroxyapatite was prepared by precipitating it from aqueous solution of Ca(NO₃)₂ and (NH₄)₂HPO₄ maintaining pH above 11. Four different methods were used for coating of hydroxyapatite on ferrite particles. SEM with EDX and X-ray diffraction analysis shows clear evidence of coating of hydroxyapatite on ferrite particles. These coated ferrite particles exhibited coercive field up to 2 kOe, which could be made useful for hysteresis heating in hyperthermia. Studies by culturing BHK-21 cells and WBC over the samples show evidence of biocompatibility. SEM micrographs and cell counts give clear indication of cell growth on the surface of the sample. Finally coated ferrite particle was implanted in Kasaulli mouse to test its biocompatibility. The magnetic properties and biocompatibility studies show that these hydroxyapatite coated ferrites could be useful for hyperthermia.     

Characterization of a novel bioactive composite using advanced X-ray computed tomography

A novel bioactive, porous silica–calcium phosphate nanocomposite (SCPC) that can be used to treat large bone defects in load-bearing positions has been tested and has shown great potential for applications in tissue engineering. Porosity is essential to the performance of the composite material as a tissue engineering scaffold, as porous scaffolds provide a physical, 3-D template to support new tissue formation. However, porosity characterization using conventional techniques such as porosimetry or scanning electron microscopy requires extensive preparation of samples and may destroy important features during preparation and analysis stage. In this work, the new composite is characterized using an advanced high resolution X-ray computed tomography, which is a non-destructive testing technique that allows construction of the 3-D topology of the microstructure. The results clearly show the effectiveness and versatility of this technique in characterizing the porous architecture of the novel composite biomaterial. The pore distribution, morphology and interconnectivity in the composite scaffolds were found to be ideal for use in tissue engineering applications.     

Properties of gas detonation ceramic coatings and their effect on the osseointegration of titanium implants for bone defect replacement

An optimal performance of bone implants with bioceramic coatings is closely related to the surface modification technology. For the first time, we have evaluated a gas detonation deposition (GDD) approach to obtain biocompatible ceramic coatings based on bioglass (BG) and calcium phosphates on Ti-based alloys as prospective materials towards their application for the development of bone implants. For the production of the coatings, hydroxyapatite (HA), HA metal-substituted (containing Ag⁺, Cu²⁺, or Zn²⁺) and tricalcium phosphate (TCP) were synthesized and characterized. Pure powders and their combination with BG were used to obtain coatings on a Ti–6Al–4V alloy using the developed automatized GDD setup. The microstructure, phase and chemical composition of the produced coatings were studied using XRD, SEM-EDS and Raman spectroscopy. The produced coated materials were evaluated in vivo in Wistar rats to analyze a reparative osteogenesis over a period of 12 weeks. The results regarding the optimization of the GDD method indicate its high productivity, as confirmed by high deposition rates. The highest deposition rate was observed for the coatings obtained from the HA metal-substituted powders. The results revealed a partial transformation of a HA phase to an α-TCP phase during the deposition, with a prevalence of the HA-phase in the coatings. According to the histological evaluation, the reparative osteogenesis occurs through the perimeter of the titanium implants, whereas the regeneration level increases from the 4th to the 12th week. The highest osteointegration level was detected for the implants coated with a biocomposite consisting of BG, HA and TCP. The results of the current study demonstrate an effectiveness of the GDD method to produce biocompatible coatings on Ti-based alloys. This provides excellent prerequisites towards the application and standardization of the GDD technology to manufacture bone implants for bone fixation and defect replacement, as well as the development of dental implants.     

Preparation and characterization of wollastonite/titanium oxide nanofiber bioceramic composite as a future implant material

In this study, novel composites consisting of electrospun titanium dioxide (TiO₂) nanofibers incorporated into high-purity wollastonite glass ceramics were prepared as materials for use in hard tissue engineering applications. These materials were characterized and investigated by means of physical, mechanical and in vitro studies. The proposed composite showed greater densification and better mechanical characteristics compared to pure wollastonite. The influence of densification temperature and TiO₂ content was investigated. Typically, TiO₂/wollastonite composites having 0, 10, 20 and 30 wt% metal oxide nanofibers were sintered at 900, 1100 and 1250 °C. The results indicated that increasing TiO₂ nanofibers content leads to increase the bulk density, compressive strength and microhardness with negligible, high and moderate influence for the densification temperature, respectively. While porosity and water adsorption capacity decreased with increasing the metal oxide nanofibers with a considerable impact for the sintering temperature in both properties. Moreover, bone-like apatite formed on the surface of wollastonite and wollastonite/TiO₂ nanofibers soaked in simulated body fluid (SBF). All these results show that the inclusion of TiO₂ nanofibers improved the characteristics of wollastonite while preserving its in vitro bioactivity; hence, the proposed composite may be used as a bone substitute in high load bearing sites.     

Direct Ink Writing of a Preceramic Polymer and Fillers to Produce Hardystonite (Ca2ZnSi2O7) Bioceramic Scaffolds

The direct ink writing of an ink composed of a preceramic polymer and fillers was used to produce hardystonite (Ca₂ZnSi₂O₇) bioceramic scaffolds. Suitable formulations were developed for the extrusion of fine filaments (350 μm diameter) through a nozzle. The preceramic polymer was employed with the double purpose of contributing to the rheology of the ink by increasing its viscosity and of forming the hardystonite phase upon heat treatment by reacting with the fillers. A control of the rheology is essential when spanning features have to be produced, and therefore the main rheological characteristics of the inks were measured (flow curves, dynamic oscillation tests, viscosity recovery tests) and compared to models reported in the literature. Highly porous scaffolds (up to 80% total porosity) were produced and heat treated in air or in nitrogen atmosphere. The influence of the heat‐treatment atmosphere on the morphology, crystalline phase assemblage, and compressive strength of the scaffolds was investigated.     

Fabrication of cancellous-bone-mimicking β-tricalcium phosphate bioceramic scaffolds with tunable architecture and mechanical strength by stereolithography 3D printing

It is highly challenging to fabricate bioceramic scaffolds mimicking architecture and mechanical strength of cancellous bone. Gyroid structure, which is based on triply periodic minimal surface, highly resembles the architecture of cancellous bone. Herein, β-tricalcium phosphate (β-TCP) bioceramic scaffolds with gyroid structure were fabricated by stereolithography (SLA) 3D printing. The SLA 3D printing ensured high precision of ceramic part. The porosity (51–87%), pore size (250 – 2400 µm), pore wall thickness (< 300 µm) and compressive strength (0.6 – 16.8 MPa) of gyroid bioceramic scaffolds were readily adjusted to match various sites of cancellous bone. The gyroid bioceramic scaffolds were more favorable for cell proliferation than the grid-like bioceramic scaffolds. The cancellous-bone-mimicking gyroid bioceramic scaffolds with tunable architecture and mechanical strength were expected to efficiently repair the target bone defects.     

Influences of gallium substitution on the phase stability,mechanical strength and cellular response of β-tricalcium phosphate bioceramics

β-tricalcium phosphate (β-TCP), a well-accepted synthetic bone grafting biomaterial, is confronted with limitations of poor phase stability and lacking the capacity to mediate the biological functions. In the current study, gallium (Ga) was substituted for calcium in the β-TCP, and the influences of Ga substitution on the phase stability, compressive strength and cellular response of β-TCP bioceramics were investigated. The results indicated that substitution of at least 2.5 mol% Ga for calcium prevented the β-TCP from transforming into α-TCP at 1250 °C. The β-TCP bioceramics substituted with 2.5 mol% Ga attained the highest compressive strength. The β-TCP bioceramics substituted with 2.5 and 5 mol% Ga showed good cytocompatibility, and suppressed in vitro osteoclastic activity as well as osteoblastic differentiation. Considering the favorable mechanical strength and the inhibitory effect on the osteoclastic activity, the β-TCP bioceramics substituted with 2.5 mol% Ga are promising for treating the bone defect in the pathological state of excessively rapid bone resorption.     

Synthesis of silicon-substituted hydroxyapatite by a hydrothermal method with two different phosphorous sources

Silicon-substituted hydroxyapatite (Si-HA) with up to 1.8 wt% Si content was prepared successfully by a hydrothermal method, using Ca(NO₃)₂, (NH₄)₃PO₄ or (NH₄)₂HPO₄ and Si(OCH₂CH₃)₄ (TEOS) as starting materials. Silicon has been incorporated in hydroxyapatite (HA) lattice by partially replacing phosphate (PO₄³⁻) groups with silicate (SiO₄⁴⁻) groups resulting in Si-HA described as Ca₁₀(PO₄)_6−x(SiO₄)_x(OH)_2−x. X-ray diffraction (XRD), Fourier transform IR spectroscopy (FTIR), inductively coupled plasma AES (ICP-AES) and scanning electron microscopy (SEM) techniques reveal that the substitution of phosphate groups by silicate groups causes some OH⁻ loss to maintain the charge balance and changes the lattice parameters of HA. The crystal shape of Si-HA has not altered compared to silicon-free reference hydroxyapatite but Si-incorporation reduces the size of Si-HA crystallites. Based on in vitro tests, soaking the specimens in simulated body fluid (SBF), and MTT assays by human osteoblast-like cells, Si-substituted hydroxyapatite is more bioactive than pure hydroxyapatite.