Surface Multifunctionalization of Inert Ceramic Implants by Calcium Phosphate Biomimetic Coating Doped with Nanoparticles Encapsulating Antibiotics.

Aseptic loosening and periprosthetic infections are complications that can occur at the interface between inert ceramic implants and natural body tissues. Therefore, the need for novel materials with antibacterial properties to prevent implant-related infection is evident. This study proposes multifunctionalizing the inert ceramic implant surface by biomimetic calcium phosphate (CaP) coating decorated with antibiotic-loaded nanoparticles for bioactivity enhancement and antibacterial effect. This study aimed to coat zirconium dioxide (ZrO2) substrates with a bioactive CaP-layer containing drug-loaded degradable polymer nanoparticles (NPs). The NPs were loaded with two antibiotics, gentamicin or bacitracin. The immobilization of NPs happened by two deposition methods: coprecipitation and drop-casting. X-ray diffraction (XRD), scanning electron microscopy (SEM), and cross-section analyses were used to characterize the coatings. MG-63 osteoblast-like cells and human mesenchymal stem cells (hMSC) were chosen for in vitro tests. Antibacterial activity was assessed with S. aureus and E. coli. The coprecipitation method allowed for a favorable homogeneous distribution of the NPs within the CaP coating. The CaP coating was constituted of hydroxyapatite and octacalcium phosphate; its thickness was 3.8 ± 1 µm with cavities of around 1 µm suitable for hosting NPs with a size of 200 nm. Antibiotics were released from the coatings in a controlled manner for 1 month. The cell culture study has confirmed the excellent behavior of the coprecipitated coating, showing cytocompatibility and a homogeneous distribution of the cells on the coated surfaces. The increase in alkaline phosphatase activity showed osteogenic differentiation. The materials were found to inhibit the growth of bacteria. Newly developed coatings with antibacterial and bioactive properties are promising candidates to prevent peri-implant infectious bone diseases.

Measurements of Density of Liquid Oxides with an Aero-Acoustic Levitator.

Densities of liquid oxide melts with melting temperatures above 2000 °C are required to establish mixing models in the liquid state for thermodynamic modeling and advanced additive manufacturing and laser welding of ceramics. Accurate measurements of molten rare earth oxide density were recently reported from experiments with an electrostatic levitator on board the International Space Station. In this work, we present an approach to terrestrial measurements of density and thermal expansion of liquid oxides from high-speed videography using an aero-acoustic levitator with laser heating and machine vision algorithms. The following density values for liquid oxides at melting temperature were obtained: Y2O3 4.6 ± 0.15; Yb2O3 8.4 ± 0.2; Zr0.9Y0.1O1.95 4.7 ± 0.2; Zr0.95Y0.05O1.975 4.9 ± 0.2; HfO2 8.2 ± 0.3 g/cm3. The accuracy of density and thermal expansion measurements can be improved by employing backlight illumination, spectropyrometry and a multi-emitter acoustic levitator.

Biomimetic in situ precipitation of calcium phosphate containing silver nanoparticles on zirconia ceramic materials for surface functionalization in terms of antimicrobial and osteoconductive properties.

OBJECTIVE: Zirconia is commonly used for manufacturing of dental implants thanks to its excellent mechanical, biological and aesthetic properties. However, its bioinertness inhibits bonding with the surrounding hard tissue and other surface interactions. In our study, we present a method for multifunctionalization of zirconia surface to improve its osseointegration and to minimize the infection risks. METHODS: For this reason, we introduced antibacterial and bioactive properties to zirconia surfaces by calcium phosphate biomimetic coating. The samples were incubated in vials in horizontal and vertical position in concentrated simulated body fluid (SBF) containing 0.1, 0.5, and 3 g/L of silver nanoparticles (Ag-NPs) and then were tested for their structure, surface properties, cytocompatibility and antibacterial properties. RESULTS AND SIGNIFICANCE: The results demonstrated that our method is suitable to introduce Ag-NPs at different concentrations into the calcium phosphate layer, i.e. from 0.05-26.6 atom% as shown by EDX. According to the results of CFU-assay these coatings exhibited antibacterial properties against S. aureus and E. coli in correlation with the concentration of Ag-NP. The potential cytotoxicity of the coated samples was determined by AlamarBlue® assay and live/dead staining of MG63 osteoblast-like cells in direct contact and by testing the extracts from the materials. Only samples containing 0.05 atom% Ag-NPs, i.e. incubated in vertical position at SBF with 0.01 g/L Ag-NPs, were found cytocompatible in direct contact with MG63 cells. On the contrary in the indirect tests, the extracts from all the materials were found cytocompatible. This method could allow developing the completely new material group, exhibiting not only one but several biological properties, which can improve osseointegration and minimize infection risks.

Toward Innovative Hemocompatible Surfaces: Crystallographic Plane Impact on Platelet Activation.

The anticoagulation treatment of cardiovascular patients, which is mandatory after implantation of heart valves or stents, has significantly adverse effects on life quality. This treatment can be reduced or even circumvented by developing novel antithrombogenic surfaces of blood-contacting implants. Thus, we aim to discover materials exhibiting outstanding hemocompatibility compared to other available synthetic materials. We present promising surficial characteristics of single crystalline alumina in terms of platelet activation inhibition. In order to elucidate the relation between its crystallographic properties including the plane orientation and blood cell behavior, we examined endothelialization, cytocompatibility, and platelet activation at the blood-alumina interfaces in a controlled experimental setup. We observed that the cell response is highly sensitive to the plane orientation and differs significantly for (0001) and (11-20) planes of Al2O3. Our results reveal for the first time the dependence of platelet activation on crystallographic orientation, which is assumed to be a critical condition controlling the thrombogenicity. Additionally, we used an endothelial cell monolayer as an internal control since endothelial cells have an impact on vessel integrity and implant acceptance. We successfully demonstrate that Al2O3(11-20) exhibits enhanced hemocompatibility in contrast to Al2O3(0001) and is comparable to the physiological endothelial monolayer in vitro.

Inkjet printed periodical micropatterns made of inert alumina ceramics induce contact guidance and stimulate osteogenic differentiation of mesenchymal stromal cells.

Bioinert high performance ceramics exhibit detrimental features for implant components with direct bone contact because of their low osseointegrating capability. We hypothesized that periodical microstructures made of inert alumina ceramics can influence the osteogenic differentiation of human mesenchymal stromal cells (hMSC). In this study, we manufactured pillared arrays made of alumina ceramics with periodicities as low as 100µm and pillar heights of 40µm employing direct inkjet printing (DIP) technique. The response of hMSC to the microstructured surfaces was monitored by measuring cell morphology, viability and formation of focal adhesion complexes. Osteogenic differentiation of hMSCs was investigated by alkaline phosphatase activity, mineralization assays and expression analysis of respective markers. We demonstrated that MSCs react to the pillars with contact guidance. Subsequently, cells grow onto and form connections between the microstructures, and at the same time are directly attached to the pillars as shown by focal adhesion stainings. Cells build up tissue-like constructs with heights up to the micropillars resulting in increased cell viability and osteogenic differentiating properties. We conclude that periodical micropatterns on the micrometer scale made of inert alumina ceramics can mediate focal adhesion dependent cell adhesion and stimulate osteogenic differentiation of hMSCs.

Calcium phosphate based three-dimensional cold plotted bone scaffolds for critical size bone defects.

Bone substitutes, like calcium phosphate, are implemented more frequently in orthopaedic surgery to reconstruct critical size defects, since autograft often results in donor site morbidity and allograft can transmit diseases. A novel bone cement, based on ß-tricalcium phosphate, polyethylene glycol, and trisodium citrate, was developed to allow the rapid manufacturing of scaffolds, by extrusion freeform fabrication, at room temperature. The cement composition exhibits good resorption properties and serves as a basis for customised (e.g., drug or growth factor loaded) scaffolds for critical size bone defects. In vitro toxicity tests confirmed proliferation and differentiation of ATDC5 cells in scaffold-conditioned culture medium. Implantation of scaffolds in the iliac wing of sheep showed bone remodelling throughout the defects, outperforming the empty defects on both mineral volume and density present in the defect after 12 weeks. Both scaffolds outperformed the autograft filled defects on mineral density, while the mineral volume present in the scaffold treated defects was at least equal to the mineral volume present in the autograft treated defects. We conclude that the formulated bone cement composition is suitable for scaffold production at room temperature and that the established scaffold material can serve as a basis for future bone substitutes to enhance de novo bone formation in critical size defects.

Calcium phosphate scaffolds mimicking the gradient architecture of native long bones.

The synthesis of beta-tricalcium phosphate (ß-TCP) scaffolds offering both the macroporous inner structure required for proper in vivo degradation and a non-macroporous outer structure for the enhancement of mechanical properties continues to be a challenge. The hypothesis of this study was to realize biomimetic ß-TCP scaffolds with a macroporous inner structure and a compact outer structure using a lost wax casting technique. The porosity, macropore size, interconnectivity of the inner porous structure, and diameter of the outer compact structure were adjusted to specific values using a three-dimensional wax printer to manufacture the wax molds for the casting process. After the slip casting, the wax was pyrolyzed and the specimens were sintered. The resulting graded ß-TCP scaffolds (porous + compact) were characterized and compared with ß-TCP scaffolds with overall apparent macropores (only porous) and samples without macropores (only compact). The porosity and the compressive strength of the only compact, porous + compact, and only porous ß-TCP samples were 31.4 ± 0.4 vol %, 55.6 ± 0.9 vol %, and 66.9 ± 0.4 vol % and 192 ± 7 MPa, 36 ± 2 MPa, and 9 ± 1 MPa, respectively. The macropore size was 500 µm and the micropore size was up to 10 µm, both featuring a completely open porous structure. From these results, we conclude that the lost wax casting technique offers an excellent method for the fabrication of ß-TCP scaffolds with an inner macroporous structure and compact outer structure which mimics the cancellous and cortical structure of natural bone.

A levitation instrument for containerless study of molten materials.

A new aero-acoustic levitation instrument (AAL) has been installed at the Institute for Mineral Engineering at RWTH University in Aachen, Germany. The AAL employs acoustically stabilized gas jet levitation with laser-beam heating and melting to create a contact-free containerless environment for high temperature materials research. Contamination-free study of liquids is possible at temperatures in excess of 3000 °C and of undercooled liquids at temperatures far below the melting point. Digital control technology advances the art of containerless experiments to obtain long-term levitation stability, allowing new experiments in extreme temperature materials research and to study operation of the levitation instrument itself. Experiments with liquid Al(2)O(3) at temperatures more than 3200 °C, 1200 °C above the melting point, and with liquid Y(3)Al(5)O(12) far below the melting point are reported. Fast pyrometry and video recording instruments yield crystallization rates in undercooled liquid Al(2)O(3) as a function of temperature. Levitation of dense liquid HfO(2) at temperatures above 2900 °C is demonstrated. Capabilities are described for resonant frequency matching in the three-axis acoustic positioning system, acoustic control of sample spin, and position control of standing wave nodes to stabilize levitation under changing experimental conditions. Further development and application of the levitation technology is discussed based on the results of experiments and modeling of instrument operations.

Manufacturing of individual biodegradable bone substitute implants using selective laser melting technique.

The additive manufacturing technique selective laser melting (SLM) has been successfully proved to be suitable for applications in implant manufacturing. SLM is well known for metal parts and offers direct manufacturing of three-dimensional (3D) parts with high bulk density on the base of individual 3D data, including computer tomography models of anatomical structures. Furthermore, an interconnecting porous structure with defined and reproducible pore size can be integrated during the design of the 3D virtual model of the implant. The objective of this study was to develop the SLM processes for a biodegradable composite material made of ß-tricalcium phosphate (ß-TCP) and poly(D, L)-lactide (PDLLA). The development of a powder composite material (ß-TCP/PDLLA) suitable for the SLM process was successfully performed. The microstructure of the manufactured samples exhibit a homogeneous arrangement of ceramic and polymer. The four-point bending strength was up to 23 MPa. The X-ray diffraction (XRD) analysis of the samples confirmed ß-TCP as the only present crystalline phase and the gel permeations chromatography (GPC) analysis documented a degradation of the polymer caused by the laser process less than conventional manufacturing processes. We conclude that SLM presents a new possibility to manufacture individual biodegradable implants made of ß-TCP/PDLLA.

Synthesis of novel tricalcium phosphate-bioactive glass composite and functionalization with rhBMP-2.

A functionalization is required for calcium phosphate-based bone substitute materials to achieve an entire bone remodeling. In this study it was hypothesized that a tailored composite of tricalcium phosphate and a bioactive glass can be loaded sufficiently with rhBMP-2 for functionalization. A composite of 40 wt% tricalcium phosphate and 60 wt% bioactive glass resulted in two crystalline phases, wollastonite and rhenanite after sintering. SEM analysis of the composite's surface revealed a spongious bone-like morphology after treatment with different acids. RhBMP-2 was immobilized non-covalently by treating with chrome sulfuric acid (CSA) and 3-aminopropyltriethoxysilane (APS) and covalently by treating with CSA/APS, and additionally with 1,1'-carbonyldiimidazole. It was proved that samples containing non-covalently immobilized rhBMP-2 on the surface exhibit significant biological activity in contrast to the samples with covalently bound protein on the surface. We conclude that a tailored composite of tricalcium phosphate and bioactive glass can be loaded sufficiently with BMP-2.

Subcritical crack growth behavior of dispersion oxide ceramics.

Zirconia (Y-TZP) is used as material for components of implants and prostheses because of its high short-term strength. The mechanical long-term reliability, however, is limited for Y-TZP because of hydrothermal aging effects and a pronounced tendency for subcritical crack growth. The hypothesis of this study was that a substantial amount of alumina in a zirconia matrix can help to significantly suppress subcritical crack growth and thereby improve the mechanical long-term reliability. The Weibull parameters as well as the parameters of the subcritical crack growth were determined for Alumina, Y-TZP, and two dispersion ceramics, that is Alumina Toughened Zirconia (ATZ, 20% alumina/80% Y-TZP), and Zirconia Toughened Alumina (ZTA, 75% alumina/25% Y-TZP). The long-term failure probability as a function of service time was predicted for the four ceramics. The parameter n of the subcritical crack growth was approx. 80% higher for ATZ compared to Y-TZP. In consequence, the estimated lifetime revealed a significant better mechanical long-term reliability for ATZ. It can be concluded that tailored dispersion oxide ceramics can address the aging problem of monolithic zirconia. This makes ATZ very interesting for components of joint replacement as well as for dental prostheses and implants.

Cytocompatibility of high strength non-oxide ceramics.

Oxide ceramic materials like alumina (Al(2)O(3)) and zirconia (ZrO(2)) are frequently used for medical applications like implants and prostheses because of their excellent biocompatibility and high wear resistance. Unfortunately, oxide ceramics cannot be used for minimal invasive thin-walled implants like resurfacing hip prostheses because of their limited strength. The hypothesis of this study is that non-oxide ceramics like silicon nitride (Si(3)N(4)) and silicon carbide (SiC)-not previously used in the medical field-are not only high strength and mechanically reliable ceramic materials due to their high amount of covalent bonds, but also exhibit a suitable biocompatibility for use as medical implants and prostheses. Mechanical investigations and cell culture tests with mouse fibroblast cells (L929) and human mesenchymal stem cells (hMSC) were performed on the ceramics. An excellent cytocompatibility was demonstrated by live/dead stainings for both L929 cells and hMSC. HMSC were able to differentiate towards osteoblasts on all tested ceramics. The determined strength of silicon nitride and silicon carbide was shown as significantly higher than that of oxide ceramics. Our results indicate that the high strength non-oxide ceramics are material candidates in the future especially for highly loaded, thin-walled implants like ceramic resurfacing hip prostheses.

Chemical strengthening of a dental lithium disilicate glass-ceramic material.

Chemical strengthening of dental ceramics by ion exchange has hitherto only been confirmed for feldspathic porcelains. The objective of this study was to examine whether the strength of lithium disilicate glass-ceramics can be increased by ion exchange as well. A lithium disilicate glass-ceramic material was treated in different molten salts. The concentration gradients of the relevant ions in the surface layer were investigated by means of electron probe microanalysis and secondary ion mass spectroscopy. Characteristic strength and Weibull modulus data were determined. An increase in strength of 25% was achieved by treatment in potassium nitrate. The chemical analyses revealed that the increase in strength resulted from an exchange of potassium for lithium ions. We conclude that ion-exchange treatments can increase the strength of lithium disilicate glass-ceramics. The improved material could be used for highly stressed applications, such as posterior crowns or inlay-retained bridges, with higher mechanical reliability.

Hemocompatibility of high strength oxide ceramic materials: an in vitro study.

High strength oxide ceramic materials like alumina and zirconia are frequently used for artificial joints because of their biocompatibility and high wear resistance. Their suitability as materials for implants and biomedical devices with direct blood contact, such as cardiovascular implants or components for blood pumps and dialyzers, has not been confirmed to date. The objective of this study was to investigate whether oxide ceramics show sufficient hemocompatibility. Dense specimens were made out of alumina, zirconia, titanium oxide, and aluminum titanate. Polyvinylchloride and silicone were additionally tested as reference materials. Interactions of human blood with the surfaces were studied by investigating partial thromboplastin time (PTT), thrombin antithrombin III complex (TAT), free plasma hemoglobin concentration, complete blood count, complement factor 5a, and protein adsorption. The results from the PTT and TAT tests clearly indicated higher blood activation by the ceramic materials when compared to the two polymer materials. However, alumina and zirconia showed lower C5a concentrations and less protein adsorption than the reference materials. Our results revealed that oxide ceramic materials alone cannot be used for implants in direct blood contact without modification of the ceramic surface, for example, by made-to-measure inert nanocoatings.

Bioactivation of inert alumina ceramics by hydroxylation.

Alumina ceramics (Al(2)O(3)) are frequently used for medical implants and prostheses because of the excellent biocompatibility, and the high mechanical reliability of the material. Inauspiciously alumina is not suitable for implant components with bone contact, because the material is bioinert and thereby no bony ongrowth, and subsequently loosening of the implant occurs. Here, we present a new method to bioactivate the surface of the material. Specimens made of high purity alumina were treated in sodium hydroxide. Cell culture tests with osteoblast-like cells as well as spectroscopical and mechanical tests were performed. Aluminium hydroxide groups were detected on the surface of the treated specimens. Enhanced cell adhesion, proliferation and secretion of osteocalcin were determined after hydroxylation. The bioactivating treatment had no deteriorating effect on the short- and long-term strength behaviour. Our results indicate that the described surface technique could be used to develop a new class of osseointegrative high-strength ceramic implants.