In Vivo Degradation Behavior of the Magnesium Alloy LANd442 in Rabbit Tibiae

In former studies the magnesium alloy LAE442 showed promising in vivo degradation behavior and biocompatibility. However, reproducibility might be enhanced by replacement of the rare earth composition metal "E" by only a single rare earth element. Therefore, it was the aim of this study to examine whether the substitution of "E" by neodymium ("Nd") had an influence on the in vivo degradation rate. LANd442 implants were inserted into rabbit tibiae and rabbits were euthanized after 4, 8, 13 and 26 weeks postoperatively. In vivo μCT was performed to evaluate the in vivo implant degradation behaviour by calculation of implant volume, density true 3-D thickness and corrosion rates. Additionally, weight loss, type of corrosion and mechanical stability were appraised by SEM/EDS-analysis and three-point bending tests. Implant volume, density and true 3-D thickness decreased over time, whereas the variance of the maximum diameters within an implant as well as the corrosion rate and weight loss increased. SEM examination revealed mainly pitting corrosion after 26 weeks. The maximum bending forces decreased over time. In comparison to LAE442, the new alloy showed a slower, but more uneven degradation behavior and less mechanical stability. To summarize, LANd442 appeared suitable for low weight bearing bones but is inferior to LAE442 regarding its degradation morphology and strength

Influence of Heat Treatment on the Degradation Behaviour of Degradable Magnesium Based Implants

Aim of the study was to characterise the influence of heat treatment on the degradation behaviour and stability of degradable magnesium based implants. For this purpose two groups (untreated/ heat treated) of LAE442 pins were separately analysed in an in vitro and in vivo study. The corrosion behaviour was evaluated during 8 weeks degradation in SBF (in vitro) and 48 weeks degradation intramedullary in the rabbit tibia (in vivo). The analyses were made by using μ-computed tomography and three-point-bending tests. Heat treatment led to altered mechanical and corrosion properties of LAE442.While the initial stability declined significantly a reduction of the degradation rate over either in vitro and in vivo evaluation period is determined. If these alterations are still reasonable for osteosynthesis implants remains to be investigated in further projects.DFG/SFB/59

In vitro impact of platinum nanoparticles on inner ear related cell culture models

So far, it was supposed that the increase of electrical impedance following cochlear implant (CI) insertion was due to technical defects of the electrode, inflammatory and/or formation of scar tissue along the electrode. However, it was recently reported that corrosion of the platinum electrode contacts may be the reason for high impedances. It could be shown that platinum particles were stripped from the electrode surfaces. Its potential cytotoxic effects within the inner ear remains to be examined. In this study in vitro cell culture models of the mouse organ of Corti cell line (HEI-OC1) and the spiral ganglion (SG) cells derived from the cochleae neonatal rats were used to investigate the effects of the polyvinylpyrrolidone coated platinum nanoparticles (Pt-NPPVP, 3 nm) on cell metabolism, neuronal survival and neurite outgrowth. Our data revealed no decrease of the metabolic activity of the HEI-OC1 cells at Pt-NPPVP concentrations between 50-150 μg/ml. Also, staining with Calcein AM/ EthD demonstrated prevalent presence of vital cells. As shown by transmission electron microscopy no Pt-NPPVP could be found at the cell surface or in the cytosol of the HEI-OC1 cells. Similarly, the SG cells exposed to 20-100 μg/ml Pt-NPPVP did not show any reduced survival rate and neurite outgrowth following staining of the neurofilament antigen even at the highest Pt-NPPVP concentration. Although the SG cells were exposed to Pt-NPPVP for further 72 h and 96 h immunocytochemical staining of the glial cells and fibroblasts presented normal cell morphology and growth independently of the cultivation period. Our data indicates that the used Pt-NPPVP do not trigger the cellular uptake and, thus, presumable do not initiate apoptotic pathways in cells of the organ of Corti cell line or the auditory nerve. The protection mechanisms to the Pt-NPPVP interactions remain to be clarified

Pressure sensing mat as an objective and sensitive tool for the evaluation of lameness in rabbits

In orthopaedic research, the analysis of the gait pattern is an often-used evaluation method. It allows an assessment of changes in motion sequence and pain level during postoperative follow up periods. Visual assessments are highly subjective and dependent on the circumstances. Particular challenge in rabbits is their hopping gait pattern. The aim of the present study was to establish a more objective and sensitive lameness evaluation using a pressure sensing mat. Twelve NZW rabbits were implemented in the study. They got an artificial anterior cruciate ligament transection of the right knee in connection with an experimental study, which investigated PTOA treatment. Rabbits were examined by a visual lameness score. Additionally, load of the hindlimbs was measured by the use of a pressure sensing mat and a video was recorded. Peak pressure and time force integral, defined as cumulated integral of all sensors associated to a hind paw, were evaluated. Preoperative data were collected on three independent days. As postoperative measurement time points, week 1 and week 12 after surgery were chosen. The subjective visual scoring was compared to the objective data of the pressure sensing mat. Following the visual score, lameness in week one was mild to moderate. In week twelve, rabbits were evaluated as lame free bar one. Contrary, following the values of the sensor mat, lameness in week one appeared to be more pronounced and almost all rabbits still showed low-grade lameness in week twelve. Consequently, the pressure sensing mat is more sensitive than the visual score and captures the grade of lameness much more accurately. For specific orthopaedic issues, where subtle differences in lameness are important to detect, the used system is a good supplementary evaluation method

Influence of PEG Chain Length of Functionalized Magnetic Nanoparticles on the Cytocompatibility and Immune Competence of Primary Murine Macrophages and Dendritic Cells

A major drawback of nanoparticles (NPs) for biomedical applications is their preferential phagocytosis in immune cells, which can be avoided by surface modifications like PEGylation. Nevertheless, examinations of different polyethylene glycol (PEG) chain lengths on the competence of immune cells as well as possible immunotoxic effects are still sparse. Therefore, primary murine macrophages and dendritic cells were generated and incubated with magnetic nanoporous silica nanoparticles (MNPSNPs) modified with different mPEG chains (2 kDa, 5 kDa, and 10 kDa). Cytotoxicity, cytokine release, and the formation of reactive oxygen species (ROS) were determined. Immune competence of both cell types was examined and uptake of MNPSNPs into macrophages was visualized. Concentrations up to 150 µg/mL MNPSNPs showed no effects on the metabolic activity or immune competence of both cell types. However, ROS significantly increased in macrophages incubated with larger PEG chains, while the concentration of cytokines (TNF-α and IL-6) did not indicate a proinflammatory process. Investigations on the uptake of MNPSNPs revealed no differences in the onset of internalization and the intensity of intracellular fluorescence. The study gives no indication for an immunotoxic effect of PEGylated MNPSNPs. Nevertheless, there is still a need for optimization regarding their internalization to ensure an efficient drug delivery

Comparison of morphological changes in efferent lymph nodes after implantation of resorbable and non-resorbable implants in rabbits

<p>Abstract</p> <p>Background</p> <p>Magnesium alloys as biodegradable implant materials received much interest in recent years. It is known that products of implant degradation can induce several types of immune response. Hence, the aim of this study was to examine the morphological changes of efferent lymph nodes after implantation of different resorbable magnesium alloys (MgCa0.8, LAE442) in comparison to commercially available resorbable (PLA) and non-resorbable (titanium) implant materials as well as control groups without implant material.</p> <p>Methods</p> <p>The different implant materials were inserted intramedullary into the rabbit tibia. After postoperative observation periods of three and six months, popliteal lymph nodes were examined histologically and immunhistologically and compared to lymph nodes of sham operated animals and animals without surgery. Haematoxylin and eosin staining was performed for cell differentiation. Mouse anti-CD79α and rat anti-CD3 monoclonal primary antibodies were used for B- and T-lymphocyte detection, mouse anti-CD68 primary antibodies for macrophage detection. Evaluation of all sections was performed applying a semi quantitative score.</p> <p>Results</p> <p>The histological evaluation demonstrated low and moderate levels of morphological changes for both magnesium alloys (LAE442 and MgCa0.8). Higher than moderate values were reached for titanium in sinus histiocytosis and histiocytic apoptosis (3 months) and for PLA in histiocytic apoptosis (3 and 6 months). The immune response to all investigated implants had a non-specific character and predominantly was a foreign-body reaction. LAE442 provoked the lowest changes which might be due to a lower degradation rate in comparison to MgCa0.8. Therewith it is a promising candidate for implants with low immunogenic potential.</p> <p>Conclusion</p> <p>Both examined magnesium alloys did not cause significantly increased morphological changes in efferent lymph nodes in comparison to the widely used implant materials titanium and PLA. LAE442 induced even lower immunological reactions. Therewith MgCa0.8 and especially LAE442 are appropriate candidates for biomedical use.</p

Evaluation of the soft tissue biocompatibility of MgCa0.8 and surgical steel 316L in vivo: a comparative study in rabbits

<p>Abstract</p> <p>Background</p> <p>Recent studies have shown the potential suitability of magnesium alloys as biodegradable implants. The aim of the present study was to compare the soft tissue biocompatibility of MgCa0.8 and commonly used surgical steel <it>in vivo</it>.</p> <p>Methods</p> <p>A biodegradable magnesium calcium alloy (MgCa0.8) and surgical steel (S316L), as a control, were investigated. Screws of identical geometrical conformation were implanted into the tibiae of 40 rabbits for a postoperative follow up of two, four, six and eight weeks. The tibialis cranialis muscle was in direct vicinity of the screw head and thus embedded in paraffin and histologically and immunohistochemically assessed. Haematoxylin and eosin staining was performed to identify macrophages, giant cells and heterophil granulocytes as well as the extent of tissue fibrosis and necrosis. Mouse anti-CD79α and rat anti-CD3 monoclonal primary antibodies were used for B- and T-lymphocyte detection. Evaluation of all sections was performed by applying a semi-quantitative score.</p> <p>Results</p> <p>Clinically, both implant materials were tolerated well. Histology revealed that a layer of fibrous tissue had formed between implant and overlying muscle in MgCa0.8 and S316L, which was demarcated by a layer of synoviocyte-like cells at its interface to the implant. In MgCa0.8 implants cavities were detected within the fibrous tissue, which were surrounded by the same kind of cell type. The thickness of the fibrous layer and the amount of tissue necrosis and cellular infiltrations gradually decreased in S316L. In contrast, a decrease could only be noted in the first weeks of implantation in MgCa0.8, whereas parameters were increasing again at the end of the observation period. B-lymphocytes were found more often in MgCa0.8 indicating humoral immunity and the presence of soluble antigens. Conversely, S316L displayed a higher quantity of T-lymphocytes.</p> <p>Conclusions</p> <p>Moderate inflammation was detected in both implant materials and resolved to a minimum during the first weeks indicating comparable biocompatibility for MgCa0.8 and S316L. Thus, the application of MgCa0.8 as biodegradable implant material seems conceivable. Since the inflammatory parameters were re-increasing at the end of the observation period in MgCa0.8 it is important to observe the development of inflammation over a longer time period in addition to the present study.</p

Increased accumulation of magnetic nanoparticles by magnetizable implant materials for the treatment of implant-associated complications

Background: In orthopaedic surgery, accumulation of agents such as anti-infectives in the bone as target tissue is difficult. The use of magnetic nanoparticles (MNPs) as carriers principally enables their accumulation via an externally applied magnetic field. Magnetizable implants are principally able to increase the strength of an externally applied magnetic field to reach also deep-seated parts in the body. Therefore, the integration of bone-addressed therapeutics in MNPs and their accumulation at a magnetic orthopaedic implant could improve the treatment of implant related infections. In this study a martensitic steel platelet as implant placeholder was used to examine its accumulation and retention capacity of MNPs in an in vitro experimental set up considering different experimental frame conditions as magnet quantity and distance to each other, implant thickness and flow velocity.Results: The magnetic field strength increased to approximately 112% when a martensitic stainless steel platelet was located between the magnet poles. Therewith a significantly higher amount of magnetic nanoparticles could be accumulated in the area of the platelet compared to the sole magnetic field. During flushing of the tube system mimicking the in vivo blood flow, the magnetized platelet was able to retain a higher amount of MNPs without an external magnetic field compared to the set up with no mounted platelet during flushing of the system. Generally, a higher flow velocity led to lower amounts of accumulated MNPs. A higher quantity of magnets and a lower distance between magnets led to a higher magnetic field strength. Albeit not significantly the magnetic field strength tended to increase with thicker platelets.Conclusion: A martensitic steel platelet significantly improved the attachment of magnetic nanoparticles in an in vitro flow system and therewith indicates the potential of magnetic implant materials in orthopaedic surgery. The use of a remanent magnetic implant material could improve the efficiency of capturing MNPs especially when the external magnetic field is turned off thus facilitating and prolonging the effect. In this way higher drug levels in the target area might be attained resulting in lower inconveniences for the patient