Uwe Gbureck

Ionic modification of calcium phosphate cement viscosity. Part II: hypodermic injection and strength improvement of brushite cement.

Brushite-forming calcium phosphate cements are of great interest as bone replacement materials because they are resorbable in physiological conditions. However, their short setting times, low mechanical strengths and limited injectability limit broad clinical application. In this study, we showed that a significant improvement of these properties of brushite cement could be achieved by the use of sodium citrate or citric acid as setting retardants, such that workable cement pastes with a powder to liquid ratio of up to 5 could be manufactured. The cement used in this study consisted of an equimolar powder mixture of beta-tricalcium phosphate and monocalcium phosphate hydrate The use of 500 mM-1M retardant solutions as liquid phase enabled initial setting times of 8-12 min. Wet compressive strength were found to be in the range between 12-18 MPa after immersion of uncompacted cement samples in serum for 24 h. A further strength improvement to 32 MPa was obtained by compaction of the cement paste during samples preparation. This is significant because high-temperature processes cannot be used to fabricate hydrated calcium phosphate materials. Cement pastes were injectable through a hypodermic needle at a powder to liquid ratio of 3.3 g/ml when a 1M citric acid was used as liquid phase, thus enabling precise controlled delivery to small defects.

Antimicrobial titanium/silver PVD coatings on titanium

BackgroundBiofilm formation and deep infection of endoprostheses is a recurrent complication in implant surgery. Post-operative infections may be overcome by adjusting antimicrobial properties of the implant surface prior to implantation. In this work we described the development of an antimicrobial titanium/silver hard coating via the physical vapor deposition (PVD) process.MethodsCoatings with a thickness of approximately 2 μm were deposited on titanium surfaces by simultaneous vaporisation of both metals in an inert argon atmosphere with a silver content of approximately 0.7 – 9% as indicated by energy dispersive X-ray analysis. On these surfaces microorganisms and eukaryotic culture cells were grown.ResultsThe coatings released sufficient silver ions (0.5–2.3 ppb) when immersed in PBS and showed significant antimicrobial potency against Staphylococcus epidermis and Klebsiella pneumoniae strains. At the same time, no cytotoxic effects of the coatings on osteoblast and epithelial cells were found.ConclusionDue to similar mechanical performance when compared to pure titanium, the TiAg coatings should be suitable to provide antimicrobial activity on load-bearing implant surfaces.

Angiogenesis in Calcium Phosphate Scaffolds by Inorganic Copper Ion Release

Angiogenesis in a tissue-engineered device may be induced by incorporating growth factors (e.g., vascular endothelial growth factor [VEGF]), genetically modified cells, and=or vascular cells. It represents an important process during the formation and repair of tissue and is essential for nourishment and supply of reparative and immunological cells. Inorganic angiogenic factors, such as copper ions, are therefore of interest in the fields of regenerative medicine and tissue engineering due to their low cost, higher stability, and potentially greater safety compared with recombinant proteins or genetic engineering approaches. The purpose of this study was to compare tissue responses to 3D printed macroporous bioceramic scaffolds implanted in mice that had been loaded with either VEGF or copper sulfate. These factors were spatially localized at the end of a single macropore some 7 mm from the surface of the scaffold. Controls without angiogenic factors exhibited only poor tissue growth within the blocks; in contrast, low doses of copper sulfate led to the formation of microvessels oriented along the macropore axis. Further, wound tissue ingrowth was particularly sensitive to the quantity of copper sulfate and was enhanced at specific concentrations or in combination with VEGF. The potential to accelerate and guide angiogenesis and wound healing by copper ion release without the expense of inductive protein(s) is highly attractive in the area of tissue-engineered bone and offers significant future potential in the field of regenerative biomaterials.

Silver-doped calcium phosphate cements with antimicrobial activity.

There is a current need for the localised delivery of antibiotics in order to treat implant-based bacterial infections. Existing treatments use non-resorbable materials such as poly(methyl methacrylate) beads loaded with antibiotics; unfortunately, as they are not resorbable, these beads require secondary surgery for removal. Calcium phosphate cements have considerable potential for the localised delivery of drugs since they can be resorbed to some extent within the body, eliminating the need for a secondary surgical procedure. Therefore, in this study, the efficacy of both hydroxyapatite and brushite cements in the delivery of silver ions has been investigated. The activity of the Ag(+) released from the cements was assessed against the growth of both Staphylococcus aureus and Staphylococcus epidermidis; the brushite cement exhibited excellent antibacterial properties and also showed an increase in compressive strength of over 30%. In this study we have found that with a few changes in Ag(+) concentration it should be possible to produce a fully resorbable bone replacement material that is combined with an antibacterial scaffold with controlled release over a period of time, which is likely to inhibit bacterial infections associated with implantation procedures.

High-strength resorbable brushite bone cement with controlled drug-releasing capabilities.

Brushite cements differ from apatite-forming compositions by consuming a lot of water in their setting reaction whereas apatite-forming cements consume little or no water at all. Only such cement systems that consume water during setting can theoretically produce near-zero porosity ceramics. This study aimed to produce such a brushite ceramic and investigated whether near elimination of porosity would prevent a burst release profile of incorporated antibiotics that is common to prior calcium phosphate cement delivery matrices. Through adjustment of the powder technological properties of the powder reactants, that is particle size and particle size distribution, and by adjusting citric acid concentration of the liquid phase to 800mM, a relative porosity of as low as 11% of the brushite cement matrix could be achieved (a 60% reduction compared to previous studies), resulting in a wet unprecompacted compressive strength of 52MPa (representing a more than 100% increase to previously reported results) with a workable setting time of 4.5min of the cement paste. Up to 2wt.% of vancomycin and ciprofloxacin could be incorporated into the cement system without loss of wet compressive strength. It was found that drug release rates could be controlled by the adjustable relative porosity of the cement system and burst release could be minimized and an almost linear release achieved, but the solubility of the antibiotic (vancomycin>ciprofloxacin) appeared also to be a crucial factor.

Binary nitride and oxynitride PVD coatings on titanium for biomedical applications

Abstract The aim of this study was the development and characterization of wear resistant and adhesive (oxy-)nitride refractory metal PVD coatings for biomedical applications. The steered arc technique with a special sector target was used to vaporize two different metal components. The subjects of investigations were layers of (Ti,Zr)N, (Ti, Nb)N and (Ti, Nb)ON with an average thickness of approximately 3–4 μm. By means of the sector target, the zirconium content of the (Ti,Zr)N layers could be varied between 50% and 70%, and the niobium content of the (Ti,Nb)N layers between 30% and 40%. X-Ray diffraction analysis showed for all materials that the cubic TiN lattice structure still dominates while a part of the titanium atoms are successively substituted by the second metal component, Zr and Nb, respectively. An excessive insertion of oxygen at interstitial sites leads to an embrittlement of the resulting coatings. All investigated coatings showed a preferred growth in the (111) direction of the cubic lattice.

Encapsulation of fibroblasts causes accelerated alginate hydrogel degradation

Calcium-alginate hydrogel has been widely studied as a material for cell encapsulation for tissue engineering. At present, the effect that cells have on the degradation of alginate hydrogel is largely unknown. We have shown that fibroblasts encapsulated at a density of 7.5 x 10(5) cells ml(-1) in both 2% and 5% w/v alginate remain viable for at least 60 days. Rheological analysis was used to study how the mechanical properties exhibited by alginate hydrogel changed during 28 days in vitro culture. Alginate degradation was shown to occur throughout the study but was greatest within the first 7 days of culture for all samples, which correlated with a sharp release of calcium ions from the construct. Fibroblasts were shown to increase the rate of degradation during the first 7 days when compared with acellular samples in both 2% and 5% w/v gels, but after 28 days both acellular and cell-encapsulating samples retained disc-shaped morphologies and gel-like spectra. The results demonstrate that although at an early stage cells influence the mechanical properties of encapsulating alginate, over a longer period of culture, the hydrogels retain sufficient mechanical integrity to exhibit gel-like properties. This allows sustained immobilization of the cells at the desired location in vivo where they can produce extracellular matrix and growth factors to expedite the healing process.

Craniofacial vertical bone augmentation: a comparison between 3D printed monolithic monetite blocks and autologous onlay grafts in the rabbit.

Onlay autografting is amongst the most predictable techniques for craniofacial vertical bone augmentation, however, complications related to donor site surgery are common and synthetic alternatives to onlay autografts are desirable. Recent studies have shown that the acidic calcium phosphates, brushite and monetite, are osteoconductive, osteoinductive and resorb faster in vivo than hydroxyapatite. Moreover, they can be 3D printed allowing precise host bone-implant conformation. The objectives of this study were to confirm that craniofacial screw fixation of 3D printed monetite blocks was possible and to compare the resulting vertical bone augmentation with autograft. 3D printed monolithic monetite onlay implants were fixed with osteosynthesis screws on the calvarial bone surface of New Zealand rabbits. After 8 weeks, integration between the implant and the calvarial bone surface was observed in all cases. Histomorphometry revealed that 42% of the monetite was resorbed and that the new bone formed within the implant occupied 43% of its volume, sufficient for immediate dental implant placement. Bone tissue within the autologous onlay occupied 60% of the volume. We observed that patterns of regeneration within the implants differed throughout the material and propose that this was due to the anatomy and blood supply pattern in the region. Rapid prototyped monetite being resorbable osteoconductive and osteoinductive would appear to be a promising biomaterial for many bone regeneration strategies.

Ion adsorption behaviour of hydroxyapatite with different crystallinities

This study aimed to correlate crystallinity of hydroxyapatite (HA) with the ion adsorption behaviour of the material. Hydroxyapatite powders of various crystallinities (X(c)) and specific surface area (SSA) were prepared by precipitation following heat treatment. Adsorption experiments were carried out by using (i) multi-component ion solutions containing a broad range of light and heavy ions to study competitive adsorption and (ii) lead and zinc solutions with concentrations up to 250 ppm to determine the adsorption isotherms of the material. While as-prepared HA powders of low crystallinity (X(c)=0%) and a high SSA of 170 m(2)/g showed quantitative removal for divalent Pb, Zn, Be, U, Bi, V, Al, Cu and Ga ions, calcined powders with higher crystallinity (X(c)=65-95%) and lower SSA between 5 and 30 m(2)/g led to a quantitative removal only for a few elements (Pb, Bi, Ga). The time and concentration dependant ion removal capacity for Pb(2+) and Zn(2+) single element solutions showed quantitative removal even after short immersion times of less than 10 min for as-prepared HA powders. XRD analysis of the powders after ion adsorption revealed the presence of pyromorphite (Pb(5)(PO(4))(3)OH) and hopeite (Zn(3)(PO(4))(2)) phases, respectively.

Cytocompatibility of brushite and monetite cell culture scaffolds made by three-dimensional powder printing

This study investigated the cytocompatibility of low-temperature direct 3-D printed calcium phosphate scaffolds in vitro. The fabrication of the scaffolds was performed with a commercial 3-D powder printing system. Diluted phosphoric acid was printed into tricalcium phosphate powder, leading to the formation of dicalcium phosphate dihydrate (brushite). Hydrothermal conversion of the brushite matrices led to the formation of dicalcium phosphate anhydrous (monetite). The biocompatibility was investigated using the osteoblastic cell line MC3T3-E1. Cell viability and the expression of alkaline phosphatase served as parameters. The culture medium was analyzed for pH value, concentration of free calcium and phosphate ions and osteocalcin. Both types of scaffolds showed a considerable increase of cell proliferation and viability; the monetite matrices were a little inferior compared with the brushite ones. The activity of alkaline phosphatase showed a similar pattern. Optical and electron microscopy revealed an obvious cell growth on the surface of both materials. Analysis of the culture medium showed minor alterations of pH value within the physiological range. The concentrations of free calcium and phosphate ions were obviously different among brushite and monetite cultures, due to their different solubility. The content of osteocalcin of the culture medium was reduced by the printed scaffolds due to adsorption. We conclude that the powder printed brushite and monetite matrices have a suitable biocompatibility for their use as cell culture scaffolds. Both materials enable osteoblastic cells in vitro to proliferate and differentiate due to the expression of typical osteoblastic markers.