Validation of a platinum bioelectrode model for preclinical electrical and biological performance evaluation.

High throughput testing of clinically representative Pt electrodes requires an inexpensive, efficient method of production. The aim of this study was to develop a facile platinum (Pt) model electrode (PME) and assess its production process, stability, and reproducibility. In this study a new model electrode was developed using representative substrates and dimensions as state-of-the-art electrode arrays used for neural stimulation. It was found that the PME is a highly reproducible robust system with similar electrochemical performance but with lower variability than other neural prosthetic arrays.Clinical Relevance- As an estimate these novel model electrodes cost 300 times less than a cochlear implant, can be manufactured in a tenth of the time and with a less than 10% failure rate. It is expected that model electrodes with low variability of electrical properties will significantly improve preclinical validation testing of electrochemical stimulation, surface modifications, and coatings.

Development and performance of a biomimetic artificial perilymph for in vitro testing of medical devices.

OBJECTIVE: Cochlear implants interface with the fluid in the cochlea called perilymph. The volume of this fluid present in human and animal model cochlea is prohibitively low for isolation for in vitro studies. Thus, there is a need for an artificial perilymph that reflects the complexity of this fluid in terms of competitive protein adsorption. APPROACH: This study established a biomimetic artificial perilymph (BAP) comprising serum albumin, immunoglobulin G, transferrin, inter-alpha-trypsin inhibitor, apolipoprotein A1 and complement C3 to represent the major components of human perilymph. Adsorption of the BAP components to platinum was analysed. MAIN RESULTS: It was established that this six component BAP provided competitive and complex adsorption behaviours consistent with biologically derived complex fluids. Additionally, adsorption of the BAP components to platinum cochlear electrodes resulted in a change in polarisation impedance consistent with that observed for the cochlear device in vivo. SIGNIFICANCE: This study established a BAP fluid suitable for furthering the understanding of the implant environment for electroactive devices that interface with the biological environment.

A critical review of cell culture strategies for modelling intracortical brain implant material reactions.

The capacity to predict in vivo responses to medical devices in humans currently relies greatly on implantation in animal models. Researchers have been striving to develop in vitro techniques that can overcome the limitations associated with in vivo approaches. This review focuses on a critical analysis of the major in vitro strategies being utilized in laboratories around the world to improve understanding of the biological performance of intracortical, brain-implanted microdevices. Of particular interest to the current review are in vitro models for studying cell responses to penetrating intracortical devices and their materials, such as electrode arrays used for brain computer interface (BCI) and deep brain stimulation electrode probes implanted through the cortex. A background on the neural interface challenge is presented, followed by discussion of relevant in vitro culture strategies and their advantages and disadvantages. Future development of 2D culture models that exhibit developmental changes capable of mimicking normal, postnatal development will form the basis for more complex accurate predictive models in the future. Although not within the scope of this review, innovations in 3D scaffold technologies and microfluidic constructs will further improve the utility of in vitro approaches.

Small bioactive molecules as dual functional co-dopants for conducting polymers.

Biological responses to neural interfacing electrodes can be modulated via biofunctionalisation of conducting polymer (CP) coatings. This study investigated the use of small bioactive molecules with anti-inflammatory properties. Specifically, anionic dexamethasone phosphate (DP) and valproic acid (VA) were used to dope the CP poly(ethylenedioxythiophene) (PEDOT). The impact of DP and VA on material properties was explored both individually and together as a codoped system, compared to the conventional dopant p-toluenesulfonate (pTS). Electrical properties of DP and VA doped PEDOT were reduced in comparison to PEDOT/pTS, however co-doping with both DP and VA was shown to significantly improve the electroactivity of PEDOT in comparison the individually doped coatings. Similarly, while the individually doped PEDOT coatings were mechanically friable, the inclusion of both dopants during electropolymerisation was shown to attenuate this response. In a whole-blood model of inflammation all DP and VA doped CPs retained their bioactivity, causing a significant reduction in levels of the pro-inflammatory cytokine TNF-α. These studies demonstrated that small charged bioactive molecules are able act as dopants for CPs and that co-doping with ions of varied size and doping affinity may provide a means of addressing the limitations of large bulky bimolecular dopants.

Performance of conducting polymer electrodes for stimulating neuroprosthetics.

OBJECTIVE: Recent interest in the use of conducting polymers (CPs) for neural stimulation electrodes has been growing; however, concerns remain regarding the stability of coatings under stimulation conditions. These studies examine the factors of the CP and implant environment that affect coating stability. The CP poly(ethylene dioxythiophene) (PEDOT) is examined in comparison to platinum (Pt), to demonstrate the potential performance of these coatings in neuroprosthetic applications. APPROACH: PEDOT is coated on Pt microelectrode arrays and assessed in vitro for charge injection limit and long-term stability under stimulation in biologically relevant electrolytes. Physical and electrical stability of coatings following ethylene oxide (ETO) sterilization is established and efficacy of PEDOT as a visual prosthesis bioelectrode is assessed in the feline model. MAIN RESULTS: It was demonstrated that PEDOT reduced the potential excursion at a Pt electrode interface by 72% in biologically relevant solutions. The charge injection limit of PEDOT for material stability was found to be on average 30× larger than Pt when tested in physiological saline and 20× larger than Pt when tested in protein supplemented media. Additionally stability of the coating was confirmed electrically and morphologically following ETO processing. It was demonstrated that PEDOT-coated electrodes had lower potential excursions in vivo and electrically evoked potentials (EEPs) could be detected within the visual cortex. SIGNIFICANCE: These studies demonstrate that PEDOT can be produced as a stable electrode coating which can be sterilized and perform effectively and safely in neuroprosthetic applications. Furthermore these findings address the necessity for characterizing in vitro properties of electrodes in biologically relevant milieu which mimic the in vivo environment more closely.

Development of sustained-release antibacterial urinary biomaterials through using an antimicrobial as an organic modifier in polyurethane nanocomposites.

Urinary catheters are among the most frequently used medical devices in clinical practice. However, their use is associated with high rates of nosocomial infection. This study investigates the use of polyurethane nanocomposites (PUNCs) incorporating an antimicrobial agent, chlorhexidine diacetate (CHX), behaving as nanoparticle dispersant and model drug/active agent, as sustained-release antibacterial biomaterials in urinary devices. A range of PUNCs incorporating organically modified silicate (OMS) nanoparticles with CHX was fabricated using a solution-cast method. PUNCs with free CHX added into the bulk polymer were also made. Materials were assessed for antibacterial activity in an in vitro urinary tract (UT) model and release kinetics of CHX was studied. PUNCs demonstrated sustained antibacterial activity against Staphylococcus epidermidis in the UT model, reaching ~50 days infection-free in materials with 2 wt % free CHX loading. Drug-release profiles demonstrated that, compared with microcomposite and unfilled polyurethane, the initial burst effect was significantly reduced in PUNCs. Prolonged drug release was achieved through incorporation of OMS, hypothesized to be due to a combination of barrier properties created by the nanoinclusions and strong interactions between CHX and MMT within the PUNCs. Use of PUNCs for sustained drug release in long-term urinary applications shows promise in addressing catheter-related nosocomial infections.

Degradable, click poly(vinyl alcohol) hydrogels: characterization of degradation and cellular compatibility.

The aim of this research was to understand the influence of functional group density on degradation and cell survival within injectable poly(vinyl alcohol) (PVA) hydrogels crosslinked through hydrazone bonds. For this purpose, PVA was modified with aldehyde and hydrazide functional groups. The click reaction between these two macromers, performed under physiologic conditions, led to hydrogel formation in less than 3 min. The influence of the crosslinking density on the gelation time, volumetric swelling ratio and mass loss of the hydrogels was investigated. These systems were slowly degradable as they maintained their gel-like state for more than 120 days. However, these networks also exhibited unusual degradation behaviour that could be the result of a breaking-forming bond phenomenon, attributable to the reversible nature of the hydrazone bond. This study also demonstrated that these networks maintained their mechanical strength while degrading, and cell encapsulation revealed the cytocompatibility of these systems.

Immunoisolating semi-permeable membranes for cell encapsulation: focus on hydrogels.

Cell-based medicine has recently emerged as a promising cure for patients suffering from various diseases and disorders that cannot be cured/treated using technologies currently available. Encapsulation within semi-permeable membranes offers transplanted cell protection from the surrounding host environment to achieve successful therapeutic function following in vivo implantation. Apart from the immunoisolation requirements, the encapsulating material must allow for cell survival and differentiation while maintaining its physico-mechanical properties throughout the required implantation period. Here we review the progress made in the development of cell encapsulation technologies from the mass transport side, highlighting the essential requirements of materials comprising immunoisolating membranes. The review will focus on hydrogels, the most common polymers used in cell encapsulation, and discuss the advantages of these materials and the challenges faced in the modification of their immunoisolating and permeability characteristics in order to optimize their function.

Conducting polymer electrodes for visual prostheses.

Conducting polymers (CPs) have the potential to provide superior neural interfaces to conventional metal electrodes by introducing more efficient charge transfer across the same geometric area. In this study the conducting polymer poly(ethylene dioxythiophene) (PEDOT) was coated on platinum (Pt) microelectrode arrays. The in vitro electrical characteristics were assessed during biphasic stimulation regimes applied between electrode pairs. It was demonstrated that PEDOT could reduce the potential excursion at a Pt electrode interface by an order of magnitude. The charge injection limit of PEDOT was found to be 15 x larger than Pt. Additionally, PEDOT coated electrodes were acutely implanted in the suprachoroidal space of a cat retina. It was demonstrated that PEDOT coated electrodes also had lower potential excursions in vivo and electrically evoked potentials (EEPs) could be detected within the vision cortex.

Antibacterial polyurethane nanocomposites using chlorhexidine diacetate as an organic modifier.

Polymer nanocomposites (NCs) are hypothesised to have enhanced barrier properties compared with pristine polymer, allowing more sustained drug release from the materials. In these NC systems active agents are typically incorporated into the polymer matrix and the release kinetics are theoretically perturbed by well dispersed nanoparticle inclusions. An alternative approach is to exploit active agent interactions with the nanoinclusion. In the proposed NC system, the driving hypothesis is that active agents can have dual functionality, acting as both drug and dispersant. Polyurethane-montmorillonite (PEU-MMT) NCs were prepared in which the antimicrobial agent chlorhexidine diacetate (CHX) was evaluated as an organic modifier for silicate dispersion. CHX was incorporated at various concentrations through organic modification of MMT or within the bulk polymer. X-ray diffraction and transmission electron microscopy analysis suggested that intercalated and partially exfoliated NCs were achieved, with better dispersion occurring in the presence of free CHX within the bulk. Tensile testing results showed that variations in the level of organic modification and nanoparticle loading modulated the mechanical properties. Material stiffness increased with nanoparticle loading relative to pristine PEU, and the ultimate properties decreased with nanoparticle and free CHX incorporation. Antibacterial activity against Staphylococcus epidermidis was significant in materials with higher exchanged MMT and NCs containing free CHX, for which 2-log reductions in adherent bacteria were found after 24h. CHX was successfully used to modulate the material properties in its dual role as a dispersant and antimicrobial agent, suggesting that alternative biocides of similar structure may behave comparably within PEU-MMT NC systems.

Novel neural interface for implant electrodes: improving electroactivity of polypyrrole through MWNT incorporation.

Multi-walled carbon nanotubes (MWNTs) can be incorporated into conductive polymers to produce superior materials for neural interfaces with high interfacial areas, conductivity and electrochemical stability. This paper explores the addition of MWNTs to polypyrrole (PPy) through two methods, layering and codeposition. Conductivity of PPy doped with polystyrene sulfonate (PSS), a commonly used dopant, was improved by 50% when MWNTs were layered with PPy/PSS. The film electrochemical stability was improved from 38% activity to 66% activity after 400 cycles of oxidation and reduction. Growth inhibition assays indicated that MWNTs are not growth inhibitory. The electroactive polymer-MWNT composites produced demonstrate properties that suggest they are promising candidates for biomedical electrode coatings.

In vitro fibroblast response to polyurethane organosilicate nanocomposites.

The term nanocomposite refers to organic:inorganic composites where one phase, typically the inorganic phase, has dimensions on the nanoscale. Several authors have noted the potential benefit of biomedical application of nanocomposite technology, and have suggested using quaternary ammonium compounds (QAC) as an organic modification to enhance dispersion of nanoparticles within polymer matrices. This study aimed to examine fibroblast responses in vitro to a range of nanocomposites using different organic modifiers. Composite materials were prepared from a polyether urethane (PEU) and various unmodified and organically modified montmorillonite (MMT) nanoparticles. QAC and amino undecanoic acid (AUA) modified-MMT were added to PEU at loadings ranging from approximately 1 to 15 wt %. Composites with organically modified QAC and AUA particles displayed partially exfoliated and intercalated silicate morphology, respectively. Nanocomposites showed increases in ultimate tensile properties for materials with lower QACMMT loadings. However QAC was shown to significantly inhibit cell growth following release from PEU-QACMMT under extraction conditions mimicking those of the physiological environment. Materials containing silicate modified using AUA were cytocompatible. The results of this study suggest that QAC may be unsuitable as organic modifiers for nanoparticles destined for biomedical use. Alternative modifiers based on AUA confer equivalent dispersion and are of low toxicity.

Chitosan adhesive for laser tissue repair: in vitro characterization.

BACKGROUND AND OBJECTIVES: Laser tissue repair usually relies on hemoderivate protein solders, based on serum albumin. These solders have intrinsic limitations that impair their widespread use, such as limited tensile strength of repaired tissue, poor solder solubility, and brittleness prior to laser denaturation. Furthermore, the required activation temperature of albumin solders (between 65 and 70 degrees C) can induce significant thermal damage to tissue. In this study, we report on the design of a new polysaccharide adhesive for tissue repair that overcomes some of the shortcomings of traditional solders. STUDY DESIGN/MATERIALS AND METHODS: Flexible and insoluble strips of chitosan adhesive (elastic modulus approximately 6.8 Mpa, surface area approximately 34 mm2, thickness approximately 20 microm) were bonded onto rectangular sections of sheep intestine using a diode laser (continuous mode, 120 +/- 10 mW, lambda = 808 nm) through a multimode optical fiber with an irradiance of approximately 15 W/cm2. The adhesive was based on chitosan and also included indocyanin green dye (IG). The temperature between tissue and adhesive was measured using a small thermocouple (diameter approximately 0.25 mm) during laser irradiation. The repaired tissue was tested for tensile strength by a calibrated tensiometer. Murine fibroblasts were cultured in extracted media from chitosan adhesive to assess cytotoxicity via cell growth inhibition in a 48 hours period. RESULTS: Chitosan adhesive successfully repaired intestine tissue, achieving a tensile strength of 14.7 +/- 4.7 kPa (mean +/- SD, n = 30) at a temperature of 60-65 degrees C. Media extracted from chitosan adhesive showed negligible toxicity to fibroblast cells under the culture conditions examined here. CONCLUSION: A novel chitosan-based adhesive has been developed, which is insoluble, flexible, and adheres firmly to tissue upon infrared laser activation.

Albumin-genipin solder for laser tissue repair.

BACKGROUND: Laser tissue soldering (LTS) is an alternative technique to suturing for tissue repair that avoids foreign body reaction and provides immediate sealing of the wound. One of the major drawbacks of LTS, however, is the weak tensile strength of the solder welds when compared to sutures. In this study, a crosslinking agent of low cytotoxicity was investigated for its ability to enhance the bond strength of albumin solders with sheep intestine. STUDY DESIGN/MATERIALS AND METHODS: Solder strips were welded onto rectangular sections of sheep small intestine using a diode laser. The laser delivered in continuous mode a power of 170 +/- 10 mW at lambda = 808 nm, through a multimode optical fiber (core size = 200 microm) to achieve a dose of 10.8 +/- 0.5 J/mg. The solder thickness and surface area were kept constant throughout the experiment (thickness = 0.15 +/- 0.01 mm, area = 12 +/- 1.2 mm2). The solder was composed of 62% bovine serum albumin (BSA), 0.38% genipin, 0.25% indocyanin green dye (IG), and water. Tissue welding was also performed with a BSA solder without genipin, as a control group. The repaired tissue was tested for tensile strength by a calibrated tensiometer. Murine fibroblasts were also cultured in extracted media from heat-denatured genipin solder to assess cell growth inhibition in a 48 hours period. RESULTS: The tensile strength of the genipin solder was doubled that of the BSA solder (0.21 +/- 0.04 N and 0.11 +/- 0.04 N, respectively; P = 10(-15) unpaired t-test, N = 30). Media extracted from crosslinked genipin solder showed negligible toxicity to fibroblast cells under the culture conditions examined here. CONCLUSION: Addition of a chemical crosslinking agent, such as genipin, significantly increased the tensile strength of adhesive-tissue bonds. A proposed mechanism for this enhanced bond strength is the synergistic action of mechanical adhesion with chemical crosslinking by genipin.

Furanones as potential anti-bacterial coatings on biomaterials.

A major barrier to the long-term use of medical devices is development of infection. Staphylococcus epidermidis is one of the most common bacterial isolates from these infections with biofilm formation being their main virulence factor. Currently, antibiotics are used as the main form of therapy. However with the emergence of staphylococcal resistance, this form of therapy is fast becoming ineffective. In this study, the ability of a novel furanone antimicrobial compound to inhibit S. epidermidis adhesion and slime production on biomaterials was assessed. Furanones were physically adsorbed to various biomaterials and bacterial load determined using radioactivity. Slime production was assessed using a colorimetric method. Additionally, the effect of the furanone coating on material surface characteristics such as hydrophobicity and surface roughness was also investigated. The results of this study indicated that there was no significant change in the material characteristics after furanone coating. Bacterial load on all furanone-coated materials was significantly reduced (p<0.001) as was slime production (p<0.001). There is a potential for furanone-coated biomaterials to be used to reduce medical device-associated infections.

Biological performance of a novel synthetic furanone-based antimicrobial.

Infection of medical devices causes significant morbidity and mortality and considerable research effort has been directed at solving this problem. The aim of this study was to assess the biological performance of a novel furanone compound that has potential as an anti-infective coating for medical devices. This study examined in vitro leukocyte response following exposure to the antibacterial 3-(1'-bromohexyl)-5-dibromomethylene-2(5H)-furanone and assessed the tissue response following subcutaneous implantation of the furanone compound covalently bound to polystyrene (PS). Peripheral human blood was exposed to furanones in solution for 1h and flow cytometry used to analyse viability and changes in expression of surface receptors CD11b/CD18 and CD44. Flow cytometry results from propidium iodide stained cell suspensions suggested that the leukocytes were viable after exposure to furanones in whole blood. No significant difference was found in the expression of CD11b/CD18 and CD44 between the furanone exposed samples and the negative control for neutrophils suggesting that the furanones themselves do not activate these leukocytes. The positive control lipopolysaccharide significantly up-regulated CD11b/CD18 and slightly down-regulated CD44 on both PMNs and monocytes. In vivo studies of the tissue response to furanone covalently bound to PS showed that there was no significant difference in cellularity of capsules surrounding the disk and no significant increase in myeloperoxidase expression. These results demonstrate negligible acute inflammatory response to synthetic brominated antibacterial furanones. Future studies will focus on chronic responses and examination of in vivo efficacy.

The control of Staphylococcus epidermidis biofilm formation and in vivo infection rates by covalently bound furanones.

In order to overcome the continuing infection rate associated with biomaterials, the use of covalently bound furanones as an antibiofilm coating for biomaterials has been investigated. Furanones have previously been shown to inhibit growth of Gram-positive and Gram-negative bacteria. The aim of these studies were to covalently bind furanones to polymers and to test their efficacy for inhibiting biofilm formation of Staphylococcus epidermidis and in vivo infection rate. Two methods of covalent attachment of furanones were used. The first, a co-polymerisation with a styrene polymer, and second, a plasma-1-ethyl-3-(dimethylaminopropyl) carbodiimide (EDC) reaction to produce furanone-coated catheters. Biofilm formation by S. epidermidis in vitro was inhibited by 89% for polystryene-furanone disks and by 78% by furanone-coated catheters (p<0.01). In an in vivo sheep model we found furanones were effective at controlling infection for up to 65 days. Furanones have potential to be used as a coating for biomaterials to control infection caused by S. epidermidis.

Compression-induced changes on physical structures and calcification of the aromatic polyether polyurethane composite.

It is generally accepted that stress causes calcification in both bio-prosthetic and polyurethane heart valves. However, simple uni-axially- and bi-axially-stretched samples did not yield a feasible model for the elaboration of the stress-induced calcification. In this study, heat compaction combined with the incorporation of polyethylene has been explored. Specimens of polyurethane were solution cast onto a porous bi-axially-drawn ultra-high-molecular-weight polyethylene film and then heat compacted under a pressure of 18 MPa at a chosen temperature for 1.5 h. The heat-compaction-induced calcification and physical changes of the polyurethane composite were evaluated using a 28-day in vitro calcification model and Attenuated Total Reflection-Fourier Transform-Infrared (ATR-FT-IR) spectroscopy. The calcification results indicated that heat-compaction-induced calcification was double that achieved without heat compaction. Heat-compacted polyurethane composite showed higher affinity to calcium ions than the non-heat compacted sample. The ATR-FT-IR results showed that the heat-compaction-induced physical changes include distortions of polymeric molecules and permanent changes of microstructures. The distortions of polymeric molecules could be deteriorated in contact with different media. The relaxation of the stressed structures of the polyether moiety might serve as a calcium trap and a heterogeneous nucleation site for calcification. The permanent changes of microstructures resulted from high distortions also served as affinity sites attracting calcification.

Fluid dynamics of a textured blood-contacting surface.

This study examined the fluid dynamics of a textured blood-contacting surface using a computational fluid-dynamic modeling technique. The texture consisted of a regular array of microfibers of length 50 or 100 microm, spaced 100 microm apart, projecting perpendicularly to the surface. The results showed that the surface texture served as a flow-retarding solid boundary for a laminar viscous flow, resulting in a lowered wall shear stress on the hase-plane surface. However, the maximum wall shear stress on the fibers was much higher than the shear stress on the nontextured phase plane. At all fractions of fiber height down past 10 microm, the permeability of the textured region greatly exceeded the analytically predictable permeability of an equivalent array of infinite-height fihers. The lowered suiface shear stress appears to explain in part the enhanced deposition of formed blood elements on the textured surface.

Performance of a polyurethane vascular prosthesis carrying a dipyridamole (Persantin) coating on its lumenal surface.

A porous polyurethane vascular prosthesis with an internal diameter of 5 mm was studied. The graft carries a coating of immobilized dipyridamole (Persantin(R)) on the surface of its lumen. Dipyridamole is a potent nontoxic inhibitor of platelet activation/aggregation, and also a strong inhibitor of vascular smooth muscle cell proliferation. The polyurethane material is also known as Chronoflex(R), and already finds use as a vascular access graft. The coated vascular graft was studied in vitro (hemocompatibility, interaction with blood platelets and cultured endothelial cells), as well as in two established in vivo models. In the first in vivo study, coated grafts were implanted in goats, as a bypass of the carotid artery (four animals, eight grafts, length of the graft was approximately 12 cm). Four uncoated grafts were used as controls in otherwise identical experiments. In the second in vivo experiment, eight sheep were used. Each animal received one coated and one uncoated prosthesis as an interposition graft in the carotid artery (length of the graft was 4 cm). The in vitro experiments revealed that the dipyridamole coating has three beneficial effects: reduced thrombogenicity, reduced adherence of blood platelets, and accommodation of a confluent monolayer of endothelial cells. The goat experiments showed patency of the coated grafts in three of the eight cases. The sheep experiments were not useful for the evaluation of the dipyridamole coating because deterioration of the polyurethane material was observed. The in vivo results indicate that the dipyridamole coating may positively influence the patency rate, probably because the coating promotes the growth of an endothelial cell lining. The sheep data show, however, that the limited stability of the Chronoflex(R) material precludes its issue for the construction of permanent small-bore vascular grafts.