Julia Kuhlmann

Revolutionizing biodegradable metals

Development of biodegradable metal implants is a complex problem because it combines engineering and medical requirements for a material. This article discusses the development of sensing and corrosion control techniques that can help in the design of biodegradable metallic implants. Biodegradable metallic implants dissolve as new tissue is formed. One of the most important factors in the design of biodegradable implants is to study the active interface, which should be monitored and controlled to address the medical concern of biocompatibility. Thus miniaturized and nanotechnology-based sensors that measure the activities of the degradation process and the formation of tissue are discussed for use with in vitro and in vivo experiments. These sensors can monitor chemical components and also cell activity and can provide new knowledge about biodegradable interfaces and how to actively control the interface to provide the best bioactivity to regenerate new tissue in a short time. Development of new alloys, nano-materials, miniature sensors, corrosion control coatings, and auxiliary applications such as biodegradable drug delivery capsules is expected to open up a new era in the engineering of materials for medicine.

Fast escape of hydrogen from gas cavities around corroding magnesium implants.

Magnesium materials are of increasing interest in the development of biodegradable implants as they exhibit properties that make them promising candidates. However, the formation of gas cavities after implantation of magnesium alloys has been widely reported in the literature. The composition of the gas and the concentration of its components in these cavities are not known as only a few studies using non-specific techniques were done about 60 years ago. Currently many researchers assume that these cavities contain primarily hydrogen because it is a product of magnesium corrosion in aqueous media. In order to clearly answer this question we implanted rare earth-containing magnesium alloy disks in mice and determined the concentration of hydrogen gas for up to 10 days using an amperometric hydrogen sensor and mass spectrometric measurements. We were able to directly monitor the hydrogen concentration over a period of 10 days and show that the gas cavities contained only a low concentration of hydrogen gas, even shortly after formation of the cavities. This means that hydrogen must be exchanged very quickly after implantation. To confirm these results hydrogen gas was directly injected subcutaneously. Most of the hydrogen gas was found to exchange within 1h after injection. Overall, our results disprove the common misbelief that these cavities mainly contain hydrogen and show how quickly this gas is exchanged with the surrounding tissue.

A system for characterizing Mg corrosion in aqueous solutions using electrochemical sensors and impedance spectroscopy

Understanding Mg corrosion is important to the development of biomedical implants made from Mg alloys. Mg corrodes readily in aqueous environments, producing H2, OH- and Mg2+. The rate of formation of these corrosion products is especially important in biomedical applications where they can affect cells and tissue near the implant. We have developed a corrosion characterization system (CCS) that allows realtime monitoring of the solution soluble corrosion products OH-, Mg2+, and H2 during immersion tests commonly used to study the corrosion of Mg materials. Instrumentation was developed to allow the system to also record electrochemical impedance spectra simultaneously in the same solution to monitor changes in the Mg samples. We demonstrated application of the CCS by observing the corrosion of Mg (99.9%) in three different corrosion solutions: NaCl, HEPES buffer, and HEPES buffer with NaCl at 37°C for 48 h. The solution concentrations of the corrosion products measured by sensors correlated with the results using standard weight loss measurements to obtain corrosion rates. This novel approach gives a better understanding of the dynamics of the corrosion process in realtime during immersion tests, rather than just providing a corrosion rate at the end of the test, and goes well beyond the immersion tests that are commonly used to study the corrosion of Mg materials. The system has the potential to be useful in systematically testing and comparing the corrosion behavior of different Mg alloys, as well as protective coatings.

In vivo monitoring the biodegradation of magnesium alloys with an electrochemical H2 sensor.

UNLABELLED Monitoring the biodegradation process of magnesium and its alloys in vivo is challenging. Currently, this process is monitored by micro-CT and X-ray imaging in vivo, which require large and costly instrumentation. Here we report a simple and effective methodology to monitor the biodegradation process in vivo by sensing H2 transdermally above a magnesium sample implanted subcutaneously in a mouse. An electrochemical H2 microsensor was used to measure the biodegradation product H2 at the surface of the skin for two magnesium alloys (ZK40 and AZ31) and one high purity magnesium single crystal (Mg8H). The sensor was able to easily detect low levels of H2 (30-400μM) permeating through the skin with a response time of about 30s. H2 levels were correlated with the biodegradation rate as determined from weight loss measurements of the implants. This new method is noninvasive, fast and requires no major equipment. STATEMENT OF SIGNIFICANCE Biomedical devices such as plates and screws used for broken bone repair are being developed out of biodegradable magnesium alloys that gradually dissolve when no longer needed. This avoids subsequent removal by surgery, which may be necessary if complications arise. A rapid, non-invasive means for monitoring the biodegradation process in vivo is needed for animal testing and point of care (POC) evaluation of patients. Here we report a novel, simple, fast, and noninvasive method to monitor the biodegradation of magnesium in vivo by measuring the biodegradation product H2 with an electrochemical H2 sensor. Since H2 rapidly permeates through biological tissue, measurements are made by simply pressing the sensor tip against the skin above the implant; the response is within 30s.

Effects of elevated magnesium and substrate on neuronal numbers and neurite outgrowth of neural stem/progenitor cells in vitro

Because a potential treatment for brain injuries could be elevating magnesium ions (Mg(2+)) intracerebrally, we characterized the effects of elevating external Mg(2+) in cultures of neonatal murine brain-derived neural stem/progenitor cells (NSCs). Using a crystal violet assay, which avoids interference of Mg(2+) in the assay, it was determined that substrate influenced Mg(2+) effects on cell numbers. On uncoated plastic, elevating Mg(2+) levels to between 2.5 and 10mM above basal increased NSC numbers, and at higher concentrations numbers decreased to control or lower levels. Similar biphasic curves were observed with different plating densities, treatment durations and length of time in culture. When cells were plated on laminin-coated plastic, NSC numbers were higher even in basal medium and no further effects were observed with Mg(2+). NSC differentiation into neurons was not altered by either substrate or Mg(2+) supplementation. Some parameters of neurite outgrowth were increased by elevated Mg(2+) when NSCs differentiated into neurons on uncoated plastic. Differentiation on laminin resulted in increased neurites even in basal medium and no further effects were seen when Mg(2+) was elevated. This system can now be used to study the multiple mechanisms by which Mg(2+) influences neuronal biology.

Protein phosphorylation studies of cerebral spinal fluid for potential biomarker development

Subarachnoid hemorrhage (SAH) followed by cerebral vasospasm (CV) leads to severe debilitation or death of an estimated one million people worldwide every year. A biomarker that would predict the onset of CV after a SAH would be useful in informing treatment protocols, but has yet to be found. The focus of this study is to explore differences in protein phosphorylation in cerebral spinal fluid (CSF) among healthy patients, SAH patients and SAH-CV patients. A significant difference in phosphorylation among the three sample types could be an important step towards the discovery of a diagnostic marker. The identification and validation of phosphorylated protein differences for study is manifested in the nature of signaling involved in the pathological events seen post SAH. Capillary liquid chromatography (cap-LC) coupled to inductively coupled plasma mass spectrometry (ICPMS) and nano-liquid chromatography-CHIP/ion trap mass spectrometry (nanoLC-CHIP/ITMS) are used to identify and measure protein phosphorylation changes in the CSF of the aforementioned groups. ICPMS represents a suitable method for screening ultra-trace phosphorus levels at the natural isotope, (31)P, while nano-LC-CHIP/ITMS is used to identify phosphoproteins by searching appropriate protein databases.

Responsive Biosensors for Biodegradable Magnesium Implants

A biosensor is an electronic device that measures biologically important parameters. An example is a sensor that measures the chemicals and materials released during corrosion of a biodegradable magnesium implant that impact surrounding cells, tissues and organs. A responsive biosensor is a biosensor that responds to its own measurements. An example is a sensor that measures the corrosion of an implant and automatically adjusts (slows down or speeds up) the corrosion rate. The University of Cincinnati, the University of Pittsburgh, North Carolina A&T State University, and the Hannover Medical Institute are collaborators in an NSF Engineering Research Center (ERC) for Revolutionizing Metallic Biomaterials (RBM). The center will use responsive sensors in experimental test beds to develop biodegradable magnesium implants. Our goal is to develop biodegradable implants that combine novel bioengineered materials based on magnesium alloys, miniature sensor devices that monitor and control the corrosion, and coatings that slow corrosion and release biological factors and drugs that will promote healing in surrounding tissues. Responsive biosensors will monitor what is happening at the interface between the implant and tissue to ensure that the implant is effective, biosafe, and provides appropriate strength while degrading. Corrosion behavior is a critical factor in the design of the implant. The corrosion behavior of implants will be studied using biosensors and through mathematical modeling. Design guidelines will be developed to predict the degradation rate of implants, and to predict and further study toxicity arising from corrosion products (i.e., Mg ion concentrations, pH levels, and hydrogen gas evolution). Knowing the corrosion rate will allow estimations to be made of implant strength and toxicity risk throughout the degradation process.Copyright

Biosensors on Enzymes, Tissues, and Cells

The first electrochemical biosensor for glucose was developed by Leland C. Clark in 1962. Driven by its huge success, fundamental and applied research has greatly expanded the concept of a biosensor since then. Today biosensors are widely used in biomedical, industrial, and environmental analysis. This chapter is focused on the fundamental concepts of biosensors and the most recent research results from electrochemical biosensors that are used for environmental analysis.

Carbon nanotube responsive materials and applications

Carbon nanotubes (CNTs) have attracted a lot of interest in the past 20 years. Superior mechanical, …