Injectable Conductive Hydrogels with Tunable Degradability as Novel Implantable Bioelectrodes.

Bioelectrodes have been developed to efficiently mediate electrical signals of biological systems as stimulators and recording devices. Recently, conductive hydrogels have garnered great attention as emerging materials for bioelectrode applications because they can permit intimate/conformal contact with living tissues and tissue-like softness. However, administration and control over the in vivo lifetime of bioelectrodes remain challenges. Here, injectable conductive hydrogels (ICHs) with tunable degradability as implantable bioelectrodes are developed. ICHs were constructed via thiol-ene reactions using poly(ethylene glycol)-tetrathiol and thiol-functionalized reduced graphene oxide with either hydrolyzable poly(ethylene glycol)-diacrylate or stable poly(ethylene glycol)-dimaleimide, the resultant hydrogels of which are degradable and nondegradable, respectively. The ICH electrodes had conductivities of 21-22 mS cm-1 and Young's moduli of 15-17 kPa, and showed excellent cell and tissue compatibility. The hydrolyzable conductive hydrogels disappeared 3 days after in vivo administration, while the stable conductive hydrogels maintained their shapes for up to 7 days. Our proof-of-concept studies reveal that electromyography signals with significantly improved sensitivity from rats could be obtained from the injected ICH electrodes compared to skin electrodes and injected nonconductive hydrogel electrodes. The ICHs, offering convenience in use, controllable degradation and excellent signal transmission, will have great potential to develop various bioelectronics devices.

Surface Characteristics of Poly(alkyl methacrylate)s from Molecular Dynamics Simulations Using All-Atom Force Field.

Molecular dynamics (MD) simulations of melt films of poly(alkyl methacrylate)s (PAMAs) with methyl, ethyl, and n-butyl substituents, respectively, have been performed using an all-atom model to investigate their surface and thin film properties. The applied all-atom force fields predict the bulk densities of PAMAs in good agreement with experiments. Moreover, predictions of the surface tensions of PMMA, PEMA, and Pn-BMA melts are in reasonably good agreement with experiments. The density profiles and orientational-order parameters of chain segments show atomic-scale characteristics in the air/polymer interfacial region. In the surface region, the backbone segments of PAMAs form a well-defined layer structure with the chain vectors oriented parallel to the surface, while the ester side-chains strongly segregate to the surface region and show perpendicular orientation to the surface, with the most pronounced surface segregation noted for Pn-BMA. Such surface segregations of chain segments make it difficult to apply a simple relationship between the cohesive energy density and the surface tension of polymers, for example, and should be taken into account in relating the surface/thin film characteristics to the bulk properties of polymers in general.

High-quantum yield alloy-typed core/shell CdSeZnS/ZnS quantum dots for bio-applications.

BACKGROUND: Quantum dots (QDs) have been used as fluorophores in various imaging fields owing to their strong fluorescent intensity, high quantum yield (QY), and narrow emission bandwidth. However, the application of QDs to bio-imaging is limited because the QY of QDs decreases substantially during the surface modification step for bio-application. RESULTS: In this study, we fabricated alloy-typed core/shell CdSeZnS/ZnS quantum dots (alloy QDs) that showed higher quantum yield and stability during the surface modification for hydrophilization compared with conventional CdSe/CdS/ZnS multilayer quantum dots (MQDs). The structure of the alloy QDs was confirmed using time-of-flight medium-energy ion scattering spectroscopy. The alloy QDs exhibited strong fluorescence and a high QY of 98.0%. After hydrophilic surface modification, the alloy QDs exhibited a QY of 84.7%, which is 1.5 times higher than that of MQDs. The QY was 77.8% after the alloy QDs were conjugated with folic acid (FA). Alloy QDs and MQDs, after conjugation with FA, were successfully used for targeting human KB cells. The alloy QDs exhibited a stronger fluorescence signal than MQD; these signals were retained in the popliteal lymph node area for 24 h. CONCLUSION: The alloy QDs maintained a higher QY in hydrophilization for biological applications than MQDs. And also, alloy QDs showed the potential as nanoprobes for highly sensitive bioimaging analysis.

Biological safety of Electroacupuncture with STS316 needles.

BACKGROUND: Electroacupuncture (EA) is often used in clinical settings due to its analgesic effect, but its safety has not been verified due to the lack of clear criteria. This study examined the critical range of the corrosion of stainless steel types STS304 and STS316, which have been used clinically, and the relationship between needle corrosion and cell necrosis. METHOD: The critical point of corrosion for STS304 and STS316 was identified by varying the time, frequency, and stimulation intensity. In a tissue necrosis experiment, EA stimulation was applied to rats using STS316 needles with different thicknesses at maximum intensity for 60 min, and the presence of corrosion and tissue necrosis was determined. A cytotoxicity experiment was also conducted and assessed the needles and tissue necrosis. RESULTS: The results showed that STS316 was more stable than STS304 and that only coated needles corroded. Furthermore, tissue necrosis was observed regardless of corrosion, and slight cell necrosis was associated with needles with corrosion. CONCLUSIONS: This study demonstrated that non-coated STS316 was the most stable for EA stimulation and that corrosion byproducts and cell necrosis were not directly related.

Enzymatically stable, non-cell adhesive, implantable polypyrrole/thiolated hyaluronic acid bioelectrodes for in vivo signal recording.

Implantable bioelectrodes have attracted significant attention for precise in vivo signal transduction with living systems. Conductive polymers, including polypyrrole (PPy), have been widely used as bioelectrodes due to their large surface areas, high charge injections, and versatilities for modification. Especially, several natural biopolymers, such as hyaluronic acid (HA), can be incorporated into conductive polymers to produce biomimetic electrodes with better biocompatibility. However, HA-incorporated PPy electrodes (PPy/HA) frequently lose their original performances after implantation in the body because of the deterioration of material properties, such as degradation of natural biopolymers in the electrode. Here, thiolated HA (HA-SH) was synthesized and introduced into PPy electrodes (PPy/HA-SH) to enhance the enzymatic stabilities of PPy electrodes against hyaluronidase (HAase) and endow these electrodes with robust resistances to non-specific cell adhesion, thereby enabling prolonged signal transmission. Unlike PPy/HA, PPy/HA-SH resisted cell adhesion even in the presence of HAase. Subcutaneous implantation studies revealed that PPy/HA-SH formed less fibrotic scar tissue and permitted more sensitive and stable signal recording for up to 15 days after implantation as compared to PPy/HA. These findings hold significance for the design and advancement of biocompatible implantable bioelectrodes for a wide range of applications, such as neural electrodes, cardiac pacemakers, and biosensors.

Size-Controllable and Monodispersed Lipid Nanoparticle Production with High mRNA Delivery Efficiency Using 3D-Printed Ring Micromixers.

Lipid nanoparticles (LNPs) are gaining recognition as potentially effective carriers for delivery of therapeutic agents, including nucleic acids (DNA and RNA), for the prevention and treatment of various diseases. Much effort has been devoted to the implementation of microfluidic techniques for the production of monodisperse and stable LNPs and the improvement of encapsulation efficiency. Here, we developed three-dimensional (3D)-printed ring micromixers for the production of size-controllable and monodispersed LNPs with a high mRNA delivery efficiency. The effects of flow rate and ring shape asymmetry on the mixing performance were initially examined. Furthermore, the physicochemical properties (such as hydrodynamic diameter, polydispersity, and encapsulation efficiency) of the generated LNPs were quantified as a function of these physical parameters via biochemical analysis and cryo-electron microscopy imaging. With a high production rate of 68 mL/min, our 3D-printed ring micromixers can be used to manufacture LNPs with diameters less than 90 nm, low polydispersity (<0.2), and high mRNA encapsulation efficiency (>91%). Despite the simplicity of the ring-shaped mixer structure, we can produce mRNA-loaded LNPs with exceptional quality and high throughput, outperforming costly commercial micromixers.

Implantable polypyrrole bioelectrodes inducing anti-inflammatory macrophage polarization for long-term in vivo signal recording.

Bioelectrodes are critical components of implantable electronic devices that enable precise electrical signal transmission in close contact with living tissues. However, their in vivo performance is often compromised by inflammatory tissue reactions mainly induced by macrophages. Hence, we aimed to develop implantable bioelectrodes with high performance and high biocompatibility by actively modulating the inflammatory response of macrophages. Consequently, we fabricated heparin-doped polypyrrole electrodes (PPy/Hep) and immobilized anti-inflammatory cytokines (interleukin-4 [IL-4]) via non-covalent interactions. IL-4 immobilization did not alter the electrochemical performance of the original PPy/Hep electrodes. In vitro primary macrophage culture revealed that IL-4-immobilized PPy/Hep electrodes induced anti-inflammatory polarization of macrophages, similar to the soluble IL-4 control. In vivo subcutaneous implantation indicated that IL-4 immobilization on PPy/Hep promoted the anti-inflammatory polarization of host macrophages and significantly mitigated scarring around the implanted electrodes. In addition, high-sensitivity electrocardiogram signals were recorded from the implanted IL-4-immobilized PPy/Hep electrodes and compared to bare gold and PPy/Hep electrodes, which were maintained for up to 15 days post-implantation. This simple and effective surface modification strategy for developing immune-compatible bioelectrodes will facilitate the development of various electronic medical devices that require high sensitivities and long-term stabilities. STATEMENT OF SIGNIFICANCE: To fabricate highly immunocompatible conductive polymer-based implantable electrodes with high performance and stability in vivo, we introduced the anti-inflammatory activity to PPy/Hep electrodes by immobilizing IL-4 via non-covalent surface modification. IL-4-immobilized PPy/Hep could significantly mitigate inflammatory responses and scarring around implants by skewing macrophages to an anti-inflammatory phenotype. The IL-4-immobilized PPy/Hep electrodes could successfully record in vivo electrocardiogram signals for up to 15 days with no substantial sensitivity loss, retaining their superior sensitivity compared to bare gold and pristine PPy/Hep electrodes. Our simple and effective surface modification strategy for developing immune-compatible bioelectrodes will facilitate the development of various electronic medical devices that require high sensitivities and long-term stabilities, such as neural electrode arrays, biosensors, and cochlear electrodes.

A Conductive and Adhesive Hydrogel Composed of MXene Nanoflakes as a Paintable Cardiac Patch for Infarcted Heart Repair.

Myocardial infarction (MI) is a major cause of death worldwide. After the occurrence of MI, the heart frequently undergoes serious pathological remodeling, leading to excessive dilation, electrical disconnection between cardiac cells, and fatal functional damage. Hence, extensive efforts have been made to suppress pathological remodeling and promote the repair of the infarcted heart. In this study, we developed a hydrogel cardiac patch that can provide mechanical support, electrical conduction, and tissue adhesiveness to aid in the recovery of an infarcted heart function. Specifically, we developed a conductive and adhesive hydrogel (CAH) by combining the two-dimensional titanium carbide (Ti3C2Tx) MXene with natural biocompatible polymers [i.e., gelatin and dextran aldehyde (dex-ald)]. The CAH was formed within 250 s of mixing the precursor solution and could be painted. The hydrogel containing 3.0 mg/mL MXene, 10% gelatin, and 5% dex-ald exhibited appropriate material characteristics for cardiac patch applications, including a uniform distribution of MXene, a high electrical conductivity (18.3 mS/cm), cardiac tissue-like elasticity (30.4 kPa), strong tissue adhesion (6.8 kPa), and resistance to various mechanical deformations. The CAH was cytocompatible and induced cardiomyocyte (CM) maturation in vitro, as indicated by the upregulation of connexin 43 expression and a faster beating rate. Furthermore, CAH could be painted onto the heart tissue and remained stably adhered to the beating epicardium. In vivo animal studies revealed that CAH cardiac patch treatment significantly improved cardiac function and alleviated the pathological remodeling of an infarcted heart. Thus, we believe that our MXene-based CAH can potentially serve as a promising platform for the effective repair of various electroactive tissues including the heart, muscle, and nerve tissues.

Biomimetic polypyrrole/hyaluronic acid electrodes integrated with hyaluronidase inhibitors offer persistent electroactivity and resistance to cell binding.

Conductive polymers, including polypyrrole (PPy), have garnered much attention as bioelectrodes because of their high conductivity, low interfacial resistance, environmental stability, and biocompatibility. In particular, the introduction of high-molecular weight hyaluronic acid (HA) into PPy enables the fabrication of biomimetic and biocompatible electrodes (i.e., PPy/HA) characterized by low biofouling. However, as HA is readily degraded by enzymes (i.e., hyaluronidase (HAase)) in a biological milieu, PPy/HA substantially loses its original properties, including resistance to cell adhesion and electrical activity. We found that HAase treatment of PPy/HA substantially degraded the HA moieties in PPy/HA, resulting in increased water contact angles, increased impedance, and conversion of non-cell adhesive to cell adhesive surfaces. Hence, it is desirable to mitigate HA degradation to achieve persistent performance of PPy/HA electrodes. Accordingly, we incorporated glycyrrhizin as an HAase inhibitor (HI) into PPy/HA electrodes. HI-incorporated PPy/HA (PPy/HA/HI) successfully prevented the degradation of the HA moiety and non-specific cell adhesion on the electrodes, in the presence of HAase (2.5 U mL-1), without cytotoxicity. These excellent properties allowed for maintenance of the electrical sensitivity of PPy/HA during cell culture with HAase. Altogether, biomimetic PPy/HA, which is resistant to degradation by HAase, may serve as an effective platform for the development of reliable and biocompatible bioelectrodes.

High-Performance Implantable Bioelectrodes with Immunocompatible Topography for Modulation of Macrophage Responses.

Implantable bioelectrodes enable precise recording or stimulation of electrical signals with living tissues in close contact. However, their performance is frequently compromised owing to inflammatory tissue reactions, which macrophages either induce or resolve by polarizing to an inflammatory (M1) or noninflammatory (M2) phenotype, respectively. Thus, we aimed to fabricate biocompatible and functional implantable conductive polymer bioelectrodes with optimal topography for the modulation of macrophage responses. To this end, we produced heparin-doped polypyrrole (PPy/Hep) electrodes of different surface roughness, with Ra values from 5.5 to 17.6 nm, by varying the charge densities during electrochemical synthesis. In vitro culture revealed that macrophages on rough PPy/Hep electrodes preferentially polarized to noninflammatory phenotypes. In particular, PPy/Hep-900 (Ra = 14 nm) was optimal with respect to electrochemical properties and the suppression of inflammatory M1 polarization. In vivo implantation indicated that PPy/Hep-900 significantly reduced macrophage recruitment, suppressed inflammatory polarization, and mitigated fibrotic tissue formation. In addition, the implanted PPy/Hep-900 electrodes could successfully record electrocardiographic signals for up to 10 days without substantial decreases in sensitivity, while other electrodes substantially lost their signal sensitivity during implantation. Altogether, we demonstrate that modulating the surface features of PPy/Hep can benefit the design and applications of high-performance and high-biocompatibility bioelectrodes.

Bcl-XL/Bax proteins direct the fate of embryonic cortical precursor cells.

In the developing mouse brain, the highest Bcl-X(L) expression is seen at the peak of neurogenesis, whereas the peak of Bax expression coincides with the astrogenic period. While such observations suggest an active role of the Bcl-2 family proteins in the generation of neurons and astrocytes, no definitive demonstration has been provided to date. Using combinations of gain- and loss-of-function assays in vivo and in vitro, we provide evidence for instructive roles of these proteins in neuronal and astrocytic fate specification. Specifically, in Bax knockout mice, astrocyte formation was decreased in the developing cortices. Overexpression of Bcl-X(L) and Bax in embryonic cortical precursors induced neural and astrocytic differentiation, respectively, while inhibitory RNAs led to the opposite results. Importantly, inhibition of caspase activity, dimerization, or mitochondrial localization of Bcl-X(L)/Bax proteins indicated that the differentiation effects of Bcl-X(L)/Bax are separable from their roles in cell survival and apoptosis. Lastly, we describe activation of intracellular signaling pathways and expression of basic helix-loop-helix transcriptional factors specific for the Bcl-2 protein-mediated differentiation.

Universal surface modification using dopamine-hyaluronic acid conjugates for anti-biofouling.

Various materials are used to make implantable devices such as pumps, catheters, and stimulators. However, such implants inevitably undergo biofouling, which is associated with non-specific proteins and cell adhesion on the material surfaces. Severe biofouling often leads to adverse tissue reactions, such as foreign body responses, and thick scar tissue formation around the implantation, which in turn hampers the function and performance of the implants. Herein, a simple and effective surface modification method using dopamine-hyaluronic acid conjugate (DA-HA) is demonstrated to implement anti-biofouling in various biomaterials. DA-HA was synthesized and utilized as a coating material and five commonly used implantable substrates (i.e., polyimide (PI), gold (Au), poly(methyl methacrylate) (PMMA), polytetrafluoroethylene (PTFE), and polyurethane (PU)) were tested for modification with DA-HA. Highly hydrophilic HA chains were immobilized on the substrate surfaces through self-polymerization of DA-HA at an alkaline pH. The DA-HA-modified substrates displayed significant resistance to both non-specific protein adsorption and cell adhesion on the surfaces. In addition, a reduction in scar tissue formation was observed in the DA-HA-coated substrates compared to the unmodified bare substrates after a 4-week subcutaneous implantation. This universal surface coating can be further implemented in various biomedical applications in which severe biofouling is highly undesirable.

Biomimetic nonbiofouling polypyrrole electrodes grafted with zwitterionic polymer using gamma rays.

Bioelectrodes, including metallic and conductive polymer (CP) bioelectrodes, often suffer from biofouling by contamination from microbacteria and/or biomolecules in biological systems, which can cause substantial impairment of biofunctionality and biocompatibility. Herein, we have employed an in situ polymerization of methacryloyloxyethyl phosphorylcholine (MPC) by gamma radiation to introduce fouling-resistant properties onto the surface of the conductive polymer, polypyrrole (PPy). The concentrations of an MPC monomer were varied during the grafting. PPy electrodes modified with MPC (PPy-g-MPC) revealed excellent anti-biofouling properties, as demonstrated by multiple analyses, such as serum protein adsorption, fibroblast adhesion, bacteria adhesion, and scar tissue formation in vivo. Importantly, PPy-g-MPC, which was modified with 0.2 M MPC using gamma radiation, exhibited electrical properties similar to unmodified PPy electrodes, indicating that our MPC grafting strategies did not cause impairment of electrical/electrochemical properties of the original PPy electrodes while successfully introducing anti-biofouling properties. Zwitterionic MPC polymer grafting on PPy electrodes by in situ polymerization with gamma radiation will benefit the development of highly biocompatible and functional bioelectrodes, such as neural electrodes, stimulators, and biosensors.

Thermodynamic interactions in double-network hydrogels.

Double-network hydrogels (DN-gels) prepared from the combination of a moderately cross-linked anionic polyelectrolyte (PE) and an uncross-linked linear polymer solution (NP) exhibit mechanical properties such as fracture toughness that are intriguingly superior to that of their individual constituents. The scheme of double-network preparation, however, is not equally successful for all polyelectrolyte/neutral polymer pairs. A successful example is the combination of poly(2-acrylamido-2-methyl-1-propane sulfonic acid) (PAMPS) cross-linked network and linear polyacrylamide (PAAm), which results in DN-gels with fracture strength under compression approaching that of articular cartilage ( approximately 20 MPa). Small-angle neutron scattering was used to determine the thermodynamic interaction parameters for PAMPS and PAAm in water as a first step to elucidate the molecular origin responsible for this superior property. Measurements on PAMPS/PAAm DN-gels and their solution blend counterparts indicate that the two polymers interact favorably with each other while in water. This favorable PAMPS/PAAm interaction given by the condition chi(PE-NP) < chi(PE-water)

Electrically Conductive Polydopamine-Polypyrrole as High Performance Biomaterials for Cell Stimulation in Vitro and Electrical Signal Recording in Vivo.

Conductive polymers (CPs) such as polypyrrole (PPY) are emerging biomaterials for use as scaffolds and bioelectrodes which interact with biological systems electrically. Still, more electrically conductive and biologically interactive CPs are required to develop high performance biomaterials and medical devices. In this study, in situ electrochemical copolymerization of polydopamine (PDA) and PPY were performed for electrode modification. Their material and biological properties were characterized using multiple techniques. The electrical properties of electrodes coated with PDA/PPY were superior to electrodes coated with PPY alone. The growth and differentiation of C2C12 myoblasts and PC12 neuronal cells on PDA/PPY was enhanced compared to PPY. Electrical stimulation of PC12 cells on PDA/PPY further promoted neuritogenesis. In vivo electromyography signal measurements demonstrated more sensitive signals from tibia muscles when using PDA/PPY-coated electrodes than bare or PPY-coated electrodes, revealing PDA/PPY to be a high-performance biomaterial with potential for various biomedical applications.

Biofiltration of a mixture of ethylene, ammonia, n-butanol, and acetone gases.

This study describes cleaning of a waste gas stream using bench scale biofilters (BFs) or biotrickling filters (BTFs). The gas stream contained a mixture of acetone, n-butanol, methane, ethylene, and ammonia, and was diverted uniformly to six biofilters and four biotrickling filters. The biofilters were packed with either perlite (BF-P), polyurethane foam (BF-F), or a mixture of compost, wood chips, and straw (BF-C), whereas the biotrickling filters contained either perlite (BTF-P) or polyurethane foam (BTF-F). Experimental results showed that both BFs and BTFs packed with various media were able to achieve complete removal of highly soluble compounds such as acetone, n-butanol, and ammonia of which the dimensionless Henry's constants (H) are less than 0.01. Methane was not removed due to its extreme insolubility (H>30). However, the ethylene (H ≈ 9) removal efficiencies depended on trickle water flow rates, media surface areas, and ammonia gas levels.

Investigation of cocaine plumes using surface acoustic wave immunoassay sensors.

Vapor sensors, aka electronic noses, are becoming an increasingly popular analytical tool for detection and identification of small molecules in the gas phase. In this paper, we present the results of a series of experiments demonstrating real-time vapor phase detection of cocaine molecules. A distinctive response or signature was observed under laboratory conditions in which the cocaine vapors were presented using an INEL vapor generator and under "field" conditions facilitated by the Georgia Bureau of Investigation (GBI) Crime Lab. For these experiments, the sensor component was a two-port resonator on ST-X quartz with a center frequency of approximately 250 MHz. On this cut of quartz, a temperature-compensated surface acoustic wave is generated via an interdigital transducer. Antibenzoylecgonine (anti-BZE) antibodies are attached to the electrodes on the device surface via a protein-A cross linker. We observed a large transient frequency shift accompanied by baseline shift with the anti-BZE coated sensor. After repeated experiments and the use of numerous controls, we believe that we have achieved real time molecular recognition of cocaine molecules.

Significance of chin-brow vertical angle in correction of kyphotic deformity of ankylosing spondylitis patients.

STUDY DESIGN: A prospective study. OBJECTIVES: To assess the significance of chin-brow vertical angle in planning and evaluating the correction of kyphotic deformity with ankylosis of the cervical spine in ankylosing spondylitis patients. SUMMARY OF BACKGROUND DATA: Accurate assessment and measurement of spinal kyphotic deformity is required when planning treatment and assessing its results. METHODS: Thirty-four ankylosing spondylitis patients with cervical ankylosis who had undergone pedicle subtraction extension osteotomy for correction of kyphotic deformity were studied. Radiographic assessment for sagittal balance was performed by measuring thoracic kyphosis, lumbar lordosis, the distance between the vertical line on the anterosuperior point of T1 and that of S1, and sacral inclination. Chin-brow vertical angle was measured on the clinical photos of the patients. Clinical outcomes were assessed by a questionnaire. RESULTS: The preoperative and postoperative chin-brow vertical angles were 35.5 degrees and 1.8 degrees, respectively. Final follow-up radiographs showed an increase in lumbar lordosis from 5.5 degrees to 43.2 degrees (an increase of 37.7 degrees ), and thoracic kyphosis remained stable from 50.4 degrees to 50.2 degrees. Sagittal imbalance significantly improved from 101.5 mm to 12.7 mm. The decreased chin-brow vertical angle correlated negatively with the correction angle. The patients with a chin-brow vertical angle of less than -10 degrees had significantly low scores on horizontal gaze. CONCLUSIONS: Chin-brow vertical angle was an objective index for evaluating horizontal gaze. Based on the results of this study, measurement of chin-brow vertical angle is recommended for planning correction of kyphosis and accurate evaluation of treatment outcome.