Bone regeneration capacity of newly developed spherical magnesium phosphate cement granules.

OBJECTIVES: Magnesium phosphate-based cements begin to catch more attention as bone substitute materials and especially as alternatives for the more commonly used calcium phosphates. In bone substitutes for augmentation purposes, atraumatic materials with good biocompatibility and resorbability are favorable. In the current study, we describe the in vivo testing of novel bone augmentation materials in form of spherical granules based on a calcium-doped magnesium phosphate (CaMgP) cement. MATERIALS AND METHODS: Granules with diameters between 500 and 710 µm were fabricated via the emulsification of CaMgP cement pastes in a lipophilic liquid. As basic material, two different CaMgP formulations were used. The obtained granules were implanted into drill hole defects at the distal femoral condyle of 27 New Zealand white rabbits for 6 and 12 weeks. After explantation, the femora were examined via X-ray diffraction analysis, histological staining, radiological examination, and EDX measurement. RESULTS: Both granule types display excellent biocompatibility without any signs of inflammation and allow for proper bone healing without the interposition of connective tissue. CaMgP granules show a fast and continuous degradation and enable fully adequate bone regeneration. CONCLUSIONS: Due to their biocompatibility, their degradation behavior, and their completely spherical morphology, these CaMgP granules present a promising bone substitute material for bone augmentation procedures, especially in sensitive areas. CLINICAL RELEVANCE: The mostly insufficient local bone supply after tooth extractions complicates prosthetic dental restoration or makes it even impossible. Therefore, bone augmentation procedures are oftentimes inevitable. Spherical CaMgP granules may represent a valuable bone replacement material in many situations.

Prefabricated and Self-Setting Cement Laminates.

Polycaprolactone (PCL) fiber mats with defined pore architecture were shown to provide sufficient support for a premixed calcium phosphate cement (CPC) paste to serve as a flat and flexible composite material for the potential application in 2-dimensional, curved cranial defects. Fiber mats were fabricated by either melt electrospinning writing (MEW) or solution electrospinning (SES) with a patterned collector. While MEW processed fiber mats led to a deterioration of the cement bending strength by approximately 50%, due to a low fiber volume content in conjunction with a weak fiber-matrix interface, fiber mats obtained by solution electrospinning resulted in a mechanical reinforcement of the cement matrix in terms of both bending strength and absorbed fracture energy. This was attributed to a higher fiber volume content and a large contact area between nanosized fibers and cement matrix. Hydrophilization of the PCL scaffolds prior to lamination further improved composite strength and preserved the comparably higher fracture energy of 1.5 to 2.0 mJ/mm². The laminate composite approach from this study was successful in demonstrating the limitations and design options of such novel composite materials. However, fiber-cement compatibility remains an issue to be addressed, since a high degree of hydrophilicity does not necessarily provoke a stronger interface.

Magnesium Phosphate Cement as Mineral Bone Adhesive.

Mineral bone cements were actually not developed for their application as bone-bonding agents, but as bone void fillers. In particular, calcium phosphate cements (CPC) are considered to be unsuitable for that application, particularly under moist conditions. Here, we showed the ex vivo ability of different magnesium phosphate cements (MPC) to adhere on bovine cortical bone substrates. The cements were obtained from a mixture of farringtonite (Mg3(PO4)2) with different amounts of phytic acid (C6H18O24P6, inositol hexaphosphate, IP6), whereas cement setting occurred by a chelation reaction between Mg2+ ions and IP6. We were able to show that cements with 25% IP6 and a powder-to-liquid ratio (PLR) of 2.0 g/mL resulted in shear strengths of 0.81 ± 0.12 MPa on bone even after 7 d storage in aqueous conditions. The samples showed a mixed adhesive-cohesive failure with cement residues on the bone surface as indicated by scanning electron microscopy and energy-dispersive X-ray analysis. The presented material demonstrated appropriate bonding characteristics, which could enable a broadening of the mineral bone cements' application field to bone adhesives.

Development and Bone Regeneration Capacity of Premixed Magnesium Phosphate Cement Pastes.

Magnesium phosphate cements (MPC) have been demonstrated to have a superior bone regeneration capacity due to their good solubility under in vivo conditions. While in the past only aqueous MPC pastes have been applied, the current study describes the fabrication and in vitro/in vivo testing of an oil-based calcium doped magnesium phosphate (CaMgP) cement paste. Premixed oil-based pastes with CaMgP chemistry combine the advantages of conventional MPC such as high mechanical strength and good resorbability with a prolonged shelf-life and an easier clinical handling. The pastes set in an aqueous environment and predominantly form struvite and achieve a compressive strength of ~8-10 MPa after setting. The implantation into a drill-hole defect at the distal femoral condyle of New Zealand white rabbits over a course of 6 and 12 weeks demonstrated good biocompatibility of the materials without the formation of soft connective tissue or any signs of inflammation. In contrast to a hydroxyapatite forming reference paste, the premixed CaMgP pastes showed subsequent degradation and bony regeneration. The CaMgP cement pastes presented herein are promising bone replacement materials with excellent material properties for an improved and facilitated clinical application.

Magnesium-based bioceramics in orthopedic applications.

Magnesium ions are directly involved in numerous biological mechanisms; for example, they play an important part in the regulation of ion channels, DNA stabilization, enzyme activation and stimulation of cell growth and proliferation. This alkaline earth metal has gained great popularity in orthopedic applications in recent years. Magnesium-based bioceramics include a large group of magnesium containing compounds such as oxides, phosphates and silicates, that are involved in orthopedic applications like bone cements, bone scaffolds or implant coatings. This article aims to give a comprehensive review on different magnesium-based bioceramics, e.g. magnesium phosphates (MgO-P2O5), calcium magnesium phosphates (CaO-MgO-P2O5), and magnesium glasses (SiO2-MgO) with a strong focus on the chemistry and properties of magnesium phosphate containing cements as the main application form. In addition, the processing of magnesium phosphate minerals into macroporous scaffolds for tissue engineering applications by either using traditional porogens or by additive manufacturing approaches are reflected. Finally, the biological in vitro and in vivo properties of magnesium phosphates for bone regeneration are summarized, which show promising results regarding the application as bone replacement material, but still lack in terms of testing in large animal models, load-bearing application sites and clinical data. STATEMENT OF SIGNIFICANCE: Though bone substitutes from calcium phosphates have been investigated for a long time, a new trend is visible in the biomaterials sector: magnesium based bioceramics from magnesium phosphates and silicates due to the special biological significance of magnesium ions in enzymatic activation, cell growth and proliferation, etc. In contrast to pure magnesium implants, such formulations do not release hydrogen during degradation. As with calcium based bioceramics, magnesium based bioceramics are used for the development of diverse applications such as cements, macroporous scaffolds and coatings. From this perspective, we present a systematic overview on diverse kinds of magnesium based bioceramics, their processing regimes for different clinical purposes and their behavior both in vitro and in vivo.

Bone regeneration capacity of magnesium phosphate cements in a large animal model.

Magnesium phosphate minerals have captured increasing attention during the past years as suitable alternatives for calcium phosphate bone replacement materials. Here, we investigated the degradation and bone regeneration capacity of experimental struvite (MgNH4PO4·6H2O) forming magnesium phosphate cements in two different orthotopic ovine implantation models. Cements formed at powder to liquid ratios (PLR) of 2.0 and 3.0 g ml-1 were implanted into trabecular bone using a non-load-bearing femoral drill-hole model and a load-bearing tibial defect model. After 4, 7 and 10 months the implants were retrieved and cement degradation and new bone formation was analyzed by micro-computed tomography (µCT) and histomorphometry. The results showed cement degradation in concert with new bone formation at both defect locations. Both cements were almost completely degraded after 10 months. The struvite cement formed with a PLR of 2.0 g ml-1 exhibited a slightly accelerated degradation kinetics compared to the cement with a PLR of 3.0 g ml-1. Tartrat-resistant acid phosphatase (TRAP) staining indicated osteoclastic resorption at the cement surface. Energy dispersive X-ray analysis (EDX) revealed that small residual cement particles were mostly accumulated in the bone marrow in between newly formed bone trabeculae. Mechanical loading did not significantly increase bone formation associated with cement degradation. Concluding, struvite-forming cements might be promising bone replacement materials due to their good degradation which is coupled with new bone formation. STATEMENT OF SIGNIFICANCE: Recently, the interest in magnesium phosphate cements (MPC) for bone substitution increased, as they exhibit high initial strength, comparably elevated degradation potential and the release of valuable magnesium ions. However, only few in vivo studies, mostly including non-load-bearing defects in small animals, have been performed to analyze the degradation and regeneration capability of MPC derived compounds. The present study examined the in vivo behavior of magnesiumammoniumphosphate hexahydrate (struvite) implants with different porosity in both mechanically loaded and non-loaded defects of merino sheep. For the first time, the effect of mechanical stimuli on the biological outcome of this clinically relevant replacement material is shown and directly compared to the conventional unloaded defect situation in a large animal model.

Mg:Ca ratio as regulating factor for osteoclastic in vitro resorption of struvite biocements.

Bioceramic degradation can occur by both passive dissolution and following active osteoclastic bone remodeling. Key parameters controlling ceramic degradation are the pH-dependent solubility product of the ceramic phase, which alters ion concentrations in physiological solution and hence regulates cell activity. This study investigated the in vitro degradation profiles of various calcium magnesium phosphate ceramics formed at low temperature. The passive resorption was measured by incubating the cement samples in cell culture medium, while active resorption was determined during a surface culture of multinuclear osteoclastic cells derived from RAW 264.7 macrophages. All surfaces showed mostly similar TRAP activities after adding RANKL-factor to stimulate osteoclastogenesis. The active degradation of the materials by osteoclasts was found to be the predominant factor for ceramic dissolution as determined by measuring the ion concentrations of cell culture medium. Here, large sized osteoclasts formed predominantly on ceramics with a Mg:Ca ratio ≥2.0 seemed to be less effective compared to smaller macrophages.

Novel bone wax based on poly(ethylene glycol)-calcium phosphate cement mixtures.

Classic bone wax is associated with drawbacks such as the risk of infection, inflammation and hindered osteogenesis. Here, we developed a novel self-setting bone wax on the basis of hydrophilic poly(ethylene glycol) (PEG) and hydroxyapatite (HA) forming calcium phosphate cement (CPC), to overcome the problems that are linked to the use of conventional beeswax systems. Amounts of up to 10 wt.% of pregelatinized starch were additionally supplemented as hemostatic agent. After exposure to a humid environment, the PEG phase dissolved and was exchanged by penetrating water that interacted with the HA precursor (tetracalcium phosphate (TTCP)/monetite) to form highly porous, nanocrystalline HA via a dissolution/precipitation reaction. Simultaneously, pregelatinized starch could gel and supply the bone wax with liquid sealing features. The novel bone wax formulation was found to be cohesive, malleable and after hardening under aqueous conditions, it had a mechanical performance (∼2.5 MPa compressive strength) that is comparable to that of cancellous bone. It withstood systolic blood pressure conditions for several days and showed antibacterial properties for almost one week, even though 60% of the incorporated drug vancomycin hydrochloride was already released after 8h of deposition by diffusion controlled processes. STATEMENT OF SIGNIFICANCE: The study investigated the development of alternative bone waxes on the basis of a hydroxyapatite (HA) forming calcium phosphate cement (CPC) system. Conventional bone waxes are composed of non-biodegradable beeswax/vaseline mixtures that are often linked to infection, inflammation and hindered osteogenesis. We combined the usage of bioresorbable polymers, the supplementation with hemostatic agents and the incorporation of a mineral component to overcome those drawbacks. Self-setting CPC precursors (tetracalcium phosphate (TTCP), monetite) were embedded in a resorbable matrix of poly(ethylene glycol) (PEG) and supplemented with pregelatinized starch. This formulation was found to be malleable and cohesive underwater. While immersion in an aqueous environment, CPC precursors formed highly porous, nanocrystalline HA via dissolution/precipitation reaction as water penetrated the novel wax formulation and PEG molecules simultaneously dissolved. The bone wax further withstood blood pressure conditions. After hardening, mechanical performance was comparable to that of cancellous bone and we also successfully provided the bone wax with antibacterial properties. In our opinion, the described bone wax formulation outmatches conventional bone waxes, as it circumvents the detriments being associated with the term "bone wax". Our wax has a novel composition and would broaden the application of CPC and besides, the general interest in bone waxes will increase, as they were long considered as a "first-line treatment" to avoid.