Improving bone defect healing using magnesium phosphate granules with tailored degradation characteristics.

OBJECTIVES: Dental implant placement frequently requires preceding bone augmentation, for example, with hydroxyapatite (HA) or ß-tricalcium phosphate (ß-TCP) granules. However, HA is degraded very slowly in vivo and for ß-TCP inconsistent degradation profiles from too rapid to rather slow are reported. To shorten the healing time before implant placement, rapidly resorbing synthetic materials are of great interest. In this study, we investigated the potential of magnesium phosphates in granular form as bone replacement materials. METHODS: Spherical granules of four different materials were prepared via an emulsion process and investigated in trabecular bone defects in sheep: struvite (MgNH4PO4·6H2O), K-struvite (MgKPO4·6H2O), farringtonite (Mg3(PO4)2) and ß-TCP. RESULTS: All materials except K-struvite exhibited promising support of bone regeneration, biomechanical properties and degradation. Struvite and ß-TCP granules degraded at a similar rate, with a relative granules area of 29% and 30% of the defect area 4 months after implantation, respectively, whereas 18% was found for farringtonite. Only the K-struvite granules degraded too rapidly, with a relative granules area of 2% remaining, resulting in initial fibrous tissue formation and intermediate impairment of biomechanical properties. SIGNIFICANCE: We demonstrated that the magnesium phosphates struvite and farringtonite have a comparable or even improved degradation behavior in vivo compared to ß-TCP. This emphasizes that magnesium phosphates may be a promising alternative to established calcium phosphate bone substitute materials.

Exploring the potential of magnesium oxychloride, an amorphous magnesium phosphate, and newberyite as possible bone cement candidates.

Magnesium phosphate-based bone cements, particularly struvite (MgNH4PO4∙6H2O)-forming cements, have attracted increased scientific interest in recent years because they exhibit similar biocompatibility to hydroxyapatite while degrading much more rapidly in vivo. However, other magnesium-based minerals which might be promising are, to date, little studied. Therefore, in this study, we investigated three magnesium-based bone cements: a magnesium oxychloride cement (Mg3(OH)5Cl∙4H2O), an amorphous magnesium phosphate cement based on Mg3(PO4)2, MgO, and NaH2PO4, and a newberyite cement (MgHPO4·3H2O). Because it is not sufficiently clear from the literature to what extent these cements are suitable for clinical use, all of them were characterized and optimized regarding setting time, setting temperature, compressive strength and passive degradation in phosphate-buffered saline. Because the in vitro properties of the newberyite cement were most promising, it was orthotopically implanted into a partially weight-bearing tibial bone defect in sheep. The cement exhibited excellent biocompatibility and degraded more rapidly compared to a hydroxyapatite reference cement; after 4 months, 18% of the cement was degraded. We conclude that the newberyite cement was the most promising candidate of the investigated cements and has clear advantages over calcium phosphate cements, especially in terms of setting time and degradation behavior.

Loss of zinc transporters ZIP1 and ZIP3 augments platelet reactivity in response to thrombin and accelerates thrombus formation in vivo.

Zinc (Zn2+) is considered as important mediator of immune cell function, thrombosis and haemostasis. However, our understanding of the transport mechanisms that regulate Zn2+ homeostasis in platelets is limited. Zn2+ transporters, ZIPs and ZnTs, are widely expressed in eukaryotic cells. Using mice globally lacking ZIP1 and ZIP3 (ZIP1/3 DKO), our aim was to explore the potential role of these Zn2+ transporters in maintaining platelet Zn2+ homeostasis and in the regulation of platelet function. While ICP-MS measurements indicated unaltered overall Zn2+ concentrations in platelets of ZIP1/3 DKO mice, we observed a significantly increased content of FluoZin3-stainable free Zn2+, which, however, appears to be released less efficiently upon thrombin-stimulated platelet activation. On the functional level, ZIP1/3 DKO platelets exhibited a hyperactive response towards threshold concentrations of G protein-coupled receptor (GPCR) agonists, while immunoreceptor tyrosine-based activation motif (ITAM)-coupled receptor agonist signalling was unaffected. This resulted in enhanced platelet aggregation towards thrombin, bigger thrombus volume under flow ex vivo and faster in vivo thrombus formation in ZIP1/3 DKO mice. Molecularly, augmented GPCR responses were accompanied by enhanced Ca2+ and PKC, CamKII and ERK1/2 signalling. The current study thereby identifies ZIP1 and ZIP3 as important regulators for the maintenance of platelet Zn2+ homeostasis and function.

Ready-To-Use and Rapidly Biodegradable Magnesium Phosphate Bone Cement: In Vivo Evaluation in Sheep.

In clinical practice, hydroxyapatite (HA) cements for bone defect treatment are frequently prepared by mixing a powder component and a liquid component shortly before implantation in the operation theater, which is time-consuming and error-prone. In addition, HA cements are only slightly resorbed, that is, cement residues can still be found in the bone years after implantation. Here, these challenges are addressed by a prefabricated magnesium phosphate cement paste based on glycerol, which is ready-to-use and can be directly applied during surgery. By using a trimodal particle size distribution (PSD), the paste is readily injectable and exhibits a compressive strength of 9-14 MPa after setting. Struvite (MgNH4 PO4 ·6H2 O), dittmarite (MgNH4 PO4 ·H2 O), farringtonite (Mg3 (PO4 )2 ), and newberyite (MgHPO4 ·3H2 O) are the mineral phases present in the set cement. The paste developed here features a promising degradation of 37% after four months in an ovine implantation model, with 25% of the implant area being newly formed bone. It is concluded that the novel prefabricated paste improves application during surgery, has a suitable degradation rate, and supports bone regeneration.

Oral Use of Therapeutic Carbon Monoxide for Anyone, Anywhere, and Anytime.

Carbon monoxide (CO) is a therapeutic gas with therapeutic potential in intestinal bowel disease. Therapeutic efficacy in the gastrointestinal tract (GIT) must be paired with safe and convenient use. Therefore, we designed an oral CO releasing system (OCORS) pairing tunable CO release into the GIT while preventing the release of any other molecule from within the device, causing safety concerns. The dimensions of the device, which is manufactured from 3D printed components, are within compendial limits. This is achieved by controlling CO decarbonylation from a molybdenum complex with a FeCl3 solution. OCORS' surrounding silicon membranes control release rates, as does the loading with carbonylated molybdenum complex and FeCl3 solution. Herein we describe the development of the system, the characterization of the CO releasing molecule (CORM), and the CO release kinetics of the overall system. Neither the CORM nor isocyanoacetate as a potential reaction byproduct were cytotoxic. Finally, we demonstrated by design validation in an in vivo porcine model that, except for the release of the therapeutic CO, OCORS isolates all components during transit through the stomach. We could show that OCORS generated and released CO locally into the stomach of the animals without systemic exposure, measured as the carboxyhemoglobin content in the blood of the pigs. In conclusion, OCORS derisks oral development by limiting patient exposure to (desirable) CO while preventing contact with any further (undesirable) chemical, by-, or degradation products. CO generating devices come in reach, which now can be used by anyone, anywhere, and anytime.

Accelerated bone regeneration through rational design of magnesium phosphate cements.

Results of several studies during past years suggested that magnesium phosphate cements (MPCs) not only show excellent biocompatibility and osteoconductivity, but they also provide improved regeneration capacity due to higher solubility compared to calcium phosphates. These findings also highlighted that chemical similarity of bone substitutes to the natural bone tissue is not a determinant factor in the success of regenerative strategies. The aim of this study was to further improve the degradation speed of MPCs for a fast bone ingrowth within a few months. We confirmed our hypothesis, that decreasing the powder-liquid ratio (PLR) of cement results in an increased content of highly soluble phases such as struvite (MgNH4PO4⋅6H2O) as well as K-struvite (MgKPO4⋅6H2O). Promising compositions with a low PLR of 1 g ml-1 were implanted in partially-loaded tibia defects in sheep. Both cements were partially degraded and replaced by bone tissue after 4 months. The degradation speed of the K-struvite cement was significantly higher compared to the struvite cement, initially resulting in the formation of a cell-rich resorption zone at the surface of some implants, as determined by histology. Overall, both MPCs investigated in this study seem to be promising as an alternative to the clinically well-established, but slowly degrading calcium phosphate cements, depending on defect size and desired degradation rate. Whereas the K-struvite cement might require further modification towards a slower resorption and reduced inflammatory response in vivo, the struvite cement appears promising for the treatment of bone defects due to its continuous degradation with simultaneous new bone formation. STATEMENT OF SIGNIFICANCE: Cold setting bone cements are used for the treatment of bone defects that exceed a critical size and cannot heal on their own. They are applied pasty into the bone defect and harden afterwards so that the shape adapts to the individual defect. Magnesium phosphates such as magnesium ammonium phosphate hexahydrate (struvite) belong to a new class of these cold setting bone cements. They degrade much faster than the clinically established calcium phosphates. In this study, a magnesium phosphate that has hardly been investigated so far was implanted into partially-loaded defects in sheeps: Potassium magnesium phosphate hexahydrate. This showed even faster resorption compared to the struvite cement: after 4 months, 63% of the cement was already degraded.

Biological and mechanical performance and degradation characteristics of calcium phosphate cements in large animals and humans.

Calcium phosphate cements (CPCs) have been used to treat bone defects and support bone regeneration because of their good biocompatibility and osteointegrative behavior. Since their introduction in the 1980s, remarkable clinical success has been achieved with these biomaterials, because they offer the unique feature of being moldable and even injectable into implant sites, where they harden through a low-temperature setting reaction. However, despite decades of research efforts, two major limitations concerning their biological and mechanical performance hamper a broader clinical use. Firstly, achieving a degradation rate that is well adjusted to the dynamics of bone formation remains a challenging issue. While apatite-forming CPCs frequently remain for years at the implant site without major signs of degradation, brushite-forming CPCs are considered to degrade to a greater extent. However, the latter tend to convert into lower soluble phases under physiological conditions, which makes their degradation behavior rather unpredictable. Secondly, CPCs exhibit insufficient mechanical properties for load bearing applications because of their inherent brittleness. This review places an emphasis on these limitations and provides an overview of studies that have investigated the biological and biomechanical performance as well as the degradation characteristics of different CPCs after implantation into trabecular bone. We reviewed studies performed in large animals, because they mimic human bone physiology more closely in terms of bone metabolism and mechanical loading conditions compared with small laboratory animals. We compared the results of these studies with clinical trials that have dealt with the degradation behavior of CPCs after vertebroplasty and kyphoplasty.

Success and side effects of different treatment options in the low current attack of bacterial biofilms on titanium implants.

The long-term success of peri-implantitis treatments is generally insufficient. Attacking the bacteria on the titanium implant surface using electrochemical polarization could be an alternative approach. In this study an E. coli biofilm in phosphate buffered saline was treated with low current densities (0.25 to 2 mA/cm2) using anodic, cathodic, or combined polarization regimes, either alone or with the antiseptic chlorhexidine. The antibacterial effect was assessed using LIVE/DEAD® staining and through quantification of viable bacteria, sample surfaces were characterized pre- and post-treatment with electrochemical impedance spectroscopy. All polarization treatments had an antibacterial effect that increased with current density, with at least 1 mA/cm2 necessary to reduce colony forming units by four orders of magnitude. Cathodic treatment was slightly superior to anodic treatment, and there was no difference between alternating polarization and single-type polarization. Neither treatment resulted in a significant detachment of bacteria, but combination with chlorhexidine improved the antibacterial effect synergistically. The use of chloride containing electrolytes is not recommended in this context. The low current densities used here were not sufficient to generate adequate bactericidal chlorine reactive species, but first signs of pitting corrosion were already detected for anodic polarization at 1 mA/cm2.

Translational control of small heat shock genes in mesophilic and thermophilic cyanobacteria by RNA thermometers.

Cyanobacteria constitute a heterogeneous phylum of oxygen-producing, photosynthetic prokaryotes. They are susceptible to various stress conditions like heat, salt, or light stress, all inducing the cyanobacterial heat shock response (HSR). Cyanobacterial small heat shock proteins (sHsps) are known to preserve thylakoid membrane integrity under stress conditions, thereby protecting the photosynthesis machinery. In Synechocystis sp PCC 6803, synthesis of the sHsp Hsp17 is regulated by an RNA thermometer (RNAT) in the 5'-untranslated region (5'-UTR) of the hsp17 mRNA. RNATs are direct temperature sensors that control expression of many bacterial heat shock and virulence genes. They hinder translation at low temperatures by base pairing, thus blocking ribosome access to the mRNA.   To explore the temperature range in which RNATs act, we studied various RNAT candidates upstream of sHsp genes from mesophilic and thermophilic cyanobacteria. The mesophilic cyanobacteria Anabaena variabilis and Nostoc sp chromosomally encode two sHsps each. Reporter gene studies suggested RNAT-mediated post-transcriptional regulation of shsp expression in both organisms. Detailed structural analysis of the two A. variabilis candidates revealed two novel RNAT types. The first, avashort, regulates translation primarily by masking of the AUG translational start codon. The second, featuring an extended initial hairpin, thus named avalong, presumably makes use of complex tertiary interaction. The 5'-UTR of the small heat shock gene hspA in the thermophile Thermosynechococcus elongatus is predicted to adopt an extended secondary structure. Structure probing revealed that the ribosome binding site was blocked at temperatures below 55 °C. The results of this study demonstrate that cyanobacteria commonly use RNATs to control expression of their small heat shock genes.