Tracy Hopkins

Poly(ε‐caprolactone)/keratin‐based composite nanofibers for biomedical applications

Keratin-based composite nanofibers have been fabricated by an electrospinning technique. Aqueous soluble keratin extracted from human hair was successfully blended with poly(ε-caprolactone) (PCL) in different ratios and transformed into nanofibrous membranes. Toward the potential use of this nanofibrous membrane in tissue engineering, its physicochemical properties, such as morphology, mechanical strength, crystallinity, chemical structure, and integrity in aqueous medium were studied and its cellular compatibility was determined. Nanofibrous membranes with PCL/keratin ratios from 100/00 to 70/30 showed good uniformity in fiber morphology and suitable mechanical properties, and retained the integrity of their fibrous structure in buffered solutions. Experimental results, using cell viability assays and scanning electron microscopy imaging, showed that the nanofibrous membranes supported 3T3 cell viability. The ability to produce blended nanofibers from protein and synthetic polymers represents a significant advancement in development of composite materials with structural and material properties that will support biomedical applications. This provides new nanofibrous materials for applications in tissue engineering and regenerative medicine.

Differential effects of antipsychotic and antidepressant drugs on neurogenic regions in rats

Increased neurogenesis in the hippocampus and subventricular zone (SVZ) of the brain of animals has been demonstrated following administration of several psychotropic medications. Such changes are thought to regenerate tissues and contribute to the beneficial effects of the medications. This study sought to determine if another neurogenic tissue, the peripheral olfactory epithelium (OE), might also exhibit changes after treatment with psychotropic medications. Young adult male rats were treated with risperidone and paliperidone, atypical antipsychotic medications; fluoxetine, a selective serotonin reuptake inhibitor (SSRI) antidepressant; and diluent control for 28days via drinking water. Bromodeoxyuridine (BrdU) was injected to label dividing cells and positive cells were quantified in the OE, cortical SVZ, and dentate gyrus (DG) of the hippocampus. In the first of two studies, paliperidone and risperidone treatment (at 1mg/kg/day) resulted in increased numbers over controls of BrdU positive cells in the OE. In the second study, examining OE, SVZ and DG in the same animal, paliperidone, but not risperidone or fluoxetine (0.6 mg/kg/day) resulted in increased cells in the OE and posterior SVZ. However, fluoxetine, but not paliperidone or risperidone treatment increased BrdU positive cells in the DG. These results show that psychotropic drug-induced cell proliferation occurs in the OE and parallels changes in the SVZ but not DG. Thus, the peripheral OE can serve as a proxy for certain psychotropic drug-induced actions on SVZ brain cell proliferation. This olfactory model can be employed in human research as a method to explore the neurogenesis effects of various pharmacologic treatments of neuropsychiatric disorders.

Initial observations on using magnesium metal in peripheral nerve repair

Biodegradable magnesium metal filaments placed inside biodegradable nerve conduits might provide the physical guidance support needed to improve the rate and extent of regeneration of peripheral nerves across injury gaps. In this study, we examined basic issues of magnesium metal resorption and biocompatibility by repairing sub-critical size gap injuries (6 mm) in one sciatic nerve of 24 adult male Lewis rats. Separated nerve stumps were connected with poly(caprolactone) nerve conduits, with and without magnesium filaments (0.25 mm diameter, 10 mm length), with two different conduit filler substances (saline and keratin hydrogel). At 6 weeks after implantation, magnesium degradation was examined by micro-computed tomography and histological analyses. Magnesium degradation was significantly greater when the conduits were filled with an acidic keratin hydrogel than with saline (p < 0.05). But magnesium filaments in some animals remained intact for 6 weeks. Using histological and immunocytochemical analyses, good biocompatibility of the magnesium implants was observed at 6 weeks, as shown by good development of regenerating nerve mini-fascicles and only mild inflammation in tissues even after complete degradation of the magnesium. Nerve regeneration was not interrupted by complete magnesium degradation. An initial functional evaluation, determination of size recovery of the gastrocnemius muscle, showed a slight improvement due to magnesium with the saline but not the keratin filler, compared with respective control conduits without magnesium. These results suggest that magnesium filament implants have the potential to improve repair of injured peripheral nerve defects in this rodent model.

Polymer Coating of Carbon Nanotube Fibers for Electric Microcables

Carbon nanotubes (CNTs) are considered the most promising candidates to replace Cu and Al in a large number of electrical, mechanical and thermal applications. Although most CNT industrial applications require macro and micro size CNT fiber assemblies, several techniques to make conducting CNT fibers, threads, yarns and ropes have been reported to this day, and improvement of their electrical and mechanical conductivity continues. Some electrical applications of these CNT conducting fibers require an insulating layer for electrical insulation and protection against mechanical tearing. Ideally, a flexible insulator such as hydrogenated nitrile butadiene rubber (HNBR) on the CNT fiber can allow fabrication of CNT coils that can be assembled into lightweight, corrosion resistant electrical motors and transformers. HNBR is a largely used commercial polymer that unlike other cable-coating polymers such as polyvinyl chloride (PVC), it provides unique continuous and uniform coating on the CNT fibers. The polymer coated/insulated CNT fibers have a 26.54 μm average diameter-which is approximately four times the diameter of a red blood cell-is produced by a simple dip-coating process. Our results confirm that HNBR in solution creates a few microns uniform insulation and mechanical protection over a CNT fiber that is used as the electrically conducting core.

Combining micro-computed tomography with histology to analyze biomedical implants for peripheral nerve repair ☆

BACKGROUND Biomedical implants used in tissue engineering repairs, such as scaffolds to repair peripheral nerves, can be too large to examine completely with histological analyses. Micro-computed tomography (micro-CT) with contrast agents allows ex vivo visualization of entire biomaterial implants and their interactions with tissues, but contrast agents can interfere with histological analyses of the tissues or cause shrinkage or loss of antigenicity. NEW METHOD Soft tissue, ex vivo micro-CT imaging using Lugols iodine was compatible with histology after using a rapid (48 h) method of removing iodine. RESULTS Adult normal and repaired rat sciatic nerves were infiltrated ex vivo with iodine, imaged with micro-CT and then the iodine was removed by incubating tissues in sodium thiosulfate. Subsequent paraffin sections of normal nerve tissues showed no differences in staining with hematoxylin and eosin or immunostaining with multiple antibodies. Iodine treatment and removal did not alter axonal diameter, nuclear size or relative area covered by immunostained axons (p>0.05). Combining imaging modalities allowed comparisons of macroscopic and microscopic features of nerve tissues regenerating through simple nerve conduits or nerve conduits containing a titanium wire for guidance. COMPARISON WITH EXISTING METHODS Quantification showed that treatment with iodine and sodium thiosulfate did not result in tissue shrinkage or loss of antigenicity. CONCLUSIONS Because this combination of treatments is rapid and does not alter tissue morphology, this expands the ex vivo methods available to examine the success of biomaterial implants used for tissue engineering repairs.

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.

An in vitro and in vivo characterization of fine WE43B magnesium wire with varied thermomechanical processing conditions

Absorbable implants made of magnesium alloys may revolutionize surgical intervention, and fine magnesium wire will be critical to many applications. Functionally, the wires must have sufficient mechanical properties to withstand implantation and in-service loading, have excellent tissue tolerance, and exhibit an appropriate degradation rate for the application. Alloy chemistry and thermomechanical processing conditions will significantly impact the materials functional performance, but the exact translation of these parameters to implant performance is unclear. With this in mind, fine (127 µm) WE43B magnesium alloy wires in five thermomechanical process (TMP) conditions (90% cold work [CW], and 250, 375, 400, and 450°C heat treatments) were investigated for their effect on mechanical and corrosion behavior. The TMP conditions gave clear metallurgical differences: transverse grain dimensions ranged from 200 nm (CW) to 3 µm (450°C), UTS varied from 324 MPa (450°C) to 608 MPa (250°C), and surgical knotting showed some were suitable (CW, 400°C, 450°C) while others were not (250°C, 350°C). In vitro and in vivo corrosion testing yielded interesting and in some cases conflicting results. After 1 month immersion in cell culture medium, wire corrosion was extensive, and TMP conditions altered the macrocorrosion morphology but not the rate or total release of magnesium ions. After 1 month subdermal implantation in mice, all wires were well tolerated and showed very little corrosion (per µCT and histology), but differences in localized corrosion were detected between conditions. This study indicates that WE43B wires treated at 450°C may be most suitable for surgical knotting procedures.

Short and long gap peripheral nerve repair with magnesium metal filaments: MAGNESIUM FILAMENTS IN PERIPHERAL NERVE REPAIR

A current clinical challenge is to replace autografts for repair of injury gaps in peripheral nerves, which can occur due to trauma or surgical interruption. Biodegradable metallic magnesium filaments, placed inside hollow nerve conduits, could support nerve repair by providing contact guidance support for axonal regeneration. This was tested by repairing sciatic nerves of adult rats with single magnesium filaments placed inside poly(caprolactone) nerve conduits. Controls were empty conduits, conduits containing titanium filaments and/or isografts from donor rats. With a nerve gap of 6 mm and 6 weeks post-repair, magnesium filaments had partially resorbed. Regenerating cells had attached to the filaments and axons were observed in distal stumps in all animals. Axon parameters were improved with magnesium compared to conduits alone or conduits with single titanium filaments. With a longer gap of 15 mm and 16 weeks post-repair, functional parameters were improved with isografts, but not with magnesium filaments or empty conduits. Magnesium filaments were completely resorbed and no evidence of scarring was seen. While axon outgrowth was not improved with the longer gap, histological measures of the tissues were improved with magnesium compared to empty conduits. Therefore, the use of magnesium filaments is promising because they are biocompatible and improve aspects of nerve regeneration.

Evaluation of peripheral nerve regeneration through biomaterial conduits via micro-CT imaging

Hollow nerve conduits made of natural or synthetic biomaterials are used clinically to aid regeneration of peripheral nerves damaged by trauma or disease. To support healing, conduit lumen patency must be maintained until recovery occurs. New methods to study conduit structural integrity would provide an important means to optimize conduits in preclinical studies. We explored a novel combined technique to examine structural integrity of two types of nerve conduits after in vivo healing.

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