Integrated insulin-iron nanoparticles: a multi-modal approach for receptor-specific bioimaging, reactive oxygen species scavenging, and wound healing.

Metallic nanoparticles have emerged as a promising option for various biological applications, owing to their distinct characteristics such as small size, optical properties, and ability to exhibit luminescence. In this study, we have successfully employed a one-pot method to synthesize multifunctional insulin-protected iron [Fe(II)] nanoparticles denoted as [IFe(II)NPs]. The formation of IFe(II)NPs is confirmed by the presence of FTIR bonds at 447.47 and 798.28 cm-1, corresponding to Fe-O and Fe-N bonds, respectively. Detailed analysis of the HR-TEM-EDS-SAED data reveals that the particles are spherical in shape, partially amorphous in nature, and have a diameter of 28.6 ± 5.2 nm. Additionally, Metal Ion Binding (MIB) and Protein Data Bank (PDB) analyses affirm the binding of iron ions to the insulin hexamer. Our findings underscore the potential of IFe(II)NPs as a promising new platform for a variety of biomedical applications due to their high signal-to-noise ratio, and minimal background fluorescence. The particles are highly luminescent, biocompatible, and have a significant quantum yield (0.632). Exemplar applications covered in this paper include insulin receptor recognition and protection against reactive oxygen species (ROS), harmful molecules known to inflict damage on cells and DNA. The IFe(II)NPs effectively mitigate ROS-induced inflammation, which is a hinderance to wound recovery, thereby facilitating enhanced wound recovery.

Edge-functionalized coal-derived graphene oxide in bacterial nanocellulose hydrogel for active wound healing.

This work presents a comparison of physicochemical and in vitro active wound healing properties of two distinct Graphene Oxides (GOs) from graphite and coal. These GOs are incorporated in Bacterial Nanocellulose (BNC) to form hydrogels. The performance and limitations of the loading fraction of both GOs in BNC are controlled by the processing technology and the source materials from which GOs are derived. Edge functionalization with C-GO offers the advantage of facilitating face-to-edge assembly in the hydrogel leading to better dispersion than the face-to-face assembly of basal functionalized G-GO. The latter leads to more aggregation of G-GO, resulting in a lower optimal loading fraction. Our investigation into the antibacterial properties of the BNC and BNC/GO hydrogels against gram-negative E. coli revealed inhibitory effects of the BNC/GO hydrogels that intensified with an increase in the concentration of GO. Furthermore, an in vitro wound scratch assay demonstrated that BNC/C-GO hydrogels promote better cell migration, confirming their superior biocompatibility and suitability as active wound dressings, albeit limited by loading fraction due to agglomeration. These findings shed light on the performance and limitations of GOs for diverse applications, emphasizing the significance of exploring the influence of different methods and source materials of GOs.

An experimental and numerical investigation of phase change electrodes for therapeutic irreversible electroporation.

Irreversible electroporation (IRE) is a new technology for ablating aberrant tissue that utilizes pulsed electric fields (PEFs) to kill cells by destabilizing their plasma membrane. When treatments are planned correctly, the pulse parameters and location of the electrodes for delivering the pulses are selected to permit destruction of the target tissue without causing thermal damage to the surrounding structures. This allows for the treatment of surgically inoperable masses that are located near major blood vessels and nerves. In select cases of high-dose IRE, where a large ablation volume is desired without increasing the number of electrode insertions, it can become challenging to design a pulse protocol that is inherently nonthermal. To solve this problem we have developed a new electrosurgical device that requires no external equipment or protocol modifications. The design incorporates a phase change material (PCM) into the electrode core that melts during treatment and absorbs heat out of the surrounding tissue. Here, this idea is reduced to practice by testing hollow electrodes filled with gallium on tissue phantoms and monitoring temperature in real time. Additionally, the experimental data generated are used to validate a numerical model of the heat transfer problem, which is then applied to investigate the cooling performance of other classes of PCMs. The results indicate that metallic PCMs, such as gallium, are better suited than organics or salt hydrates for thermal management, because their comparatively higher thermal conductivity aids in heat dissipation. However, the melting point of the metallic PCM must be properly adjusted to ensure that the phase transition is not completed before the end of treatment. When translated clinically, phase change electrodes have the potential to continue to allow IRE to be performed safely near critical structures, even in high-dose cases.

Therapeutic potential of lactoferrin-coated iron oxide nanospheres for targeted hyperthermia in gastric cancer.

Lactoferrin (LF) is a non-heme iron-binding glycoprotein involved in the transport of iron in blood plasma. In addition, it has many biological functions, including antibacterial, antiviral, antimicrobial, antiparasitic, and, importantly, antitumor properties. In this study, we have investigated the potential of employing lactoferrin-iron oxide nanoparticles (LF-IONPs) as a treatment modality for gastric cancer. The study confirms the formation of LF-IONPs with a spherical shape and an average size of 5 ± 2 nm, embedded within the protein matrix. FTIR and Raman analysis revealed that the Fe-O bond stabilized the protein particle interactions. Further, we conducted hyperthermia studies to ascertain whether the proposed composite can generate a sufficient rise in temperature at a low frequency. The results confirmed that we can achieve a temperature rise of about 7 °C at 242.4 kHz, which can be further harnessed for gastric cancer treatment. The particles were further tested for their anti-cancer activity on AGS cells, with and without hyperthermia. Results indicate that LF-IONPs (10 µg/ml) significantly enhance cytotoxicity, resulting in the demise of 67.75 ± 5.2% of cells post hyperthermia, while also exhibiting an inhibitory effect on cell migration compared to control cells, with the most inhibition observed after 36 h of treatment. These findings suggest the potential of LF-IONPs in targeted hyperthermia treatment of gastric cancer.

Determination of Hydrophobic Dispersive Surface Free Energy of Activated Carbon Fibers Measured by Inverse Gas Chromatographic Technique.

Activated carbon fibers (ACFs) as one of the most important porous carbon materials are widely used in many applications that involve rapid adsorption and low-pressure loss, including air purification, water treatment, and electrochemical applications. For designing such fibers for the adsorption bed in gas and aqueous phases, in-depth comprehension of the surface components is crucial. However, achieving reliable values remains a major challenge due to the high adsorption affinity of ACFs. To overcome this problem, we propose a novel approach to determine London dispersive components (γSL) of the surface free energy of ACFs by inverse gas chromatography (IGC) technique at an infinite dilution. Our data reveal the γSL values at 298 K for bare carbon fibers (CFs) and the ACFs to be 97 and 260-285 mJ·m-2, respectively, which lie in the regime of secondary bonding of physical adsorption. Our analysis indicates that these are impacted by micropores and defects on the carbon surfaces. Comparing the γSL obtained by the traditional Gray's method, our method is concluded as the most accurate and reliable value for the hydrophobic dispersive surface component of porous carbonaceous materials. As such, it could serve as a valuable tool in designing interface engineering in adsorption-related applications.

Surface energetics of graphene oxide and reduced graphene oxide determined by inverse gas chromatographic technique at infinite dilution at room temperature.

Graphene is of great interest for many far-reaching applications that involve interparticle interactions in adsorbents, coatings, and composites. A deep understanding of the surface components has been crucial but achieving the most accurate and reliable values of these, unaffected by experimental conditions or the analytical techniques used, remains a major challenge. To this end, we have proposed in this paper a novel approach for the first time, to the best of our knowledge, to determine London dispersive and specific (polar) components including the Lewis acid-base character of the surface free energy of graphene materials (graphene oxide (GO), reduced graphene oxide (rGO), and graphite) using inverse gas chromatography (IGC) technique at an infinite dilution. We have estimated the London dispersive surface energy values of graphite, GO, and rGO as van der Waals interaction to be 156-179, 89-106, and 110-119 mJ m-2, respectively, in the temperature range of 320-360 K. These are attributable to the surface properties impacted by the oxygen moieties, defects, and micropores on the carbon frameworks. Further, the acceptor (KA) and donor (KD) parameters of GO were found to be 0.71 and 0.96, respectively, while those of rGO were 0.54 and 1.05. Notably, the GO is more of the Lewis acid character that could be amphoteric, while the Lewis base characteristics of both GO and rGO are not significantly changed. These results provide foundational knowledge to understand the physicochemical properties of graphene surfaces, which should be helpful to designing interface engineering in various applications.

Bacterioboat-A novel tool to increase the half-life period of the orally administered drug.

The short half-life in the GI tract necessitates an excess of drugs causing side effects of oral formulations. Here, we report the development and deployment of Bacterioboat, which consists of surface-encapsulated mesoporous nanoparticles on metabolically active Lactobacillus reuteri as a drug carrier suitable for oral administration. Bacterioboat showed up to 16% drug loading of its dry weight, intestinal anchorage around alveoli regions, sustained release, and stability in physiological conditions up to 24 hours. In vivo studies showed that oral delivery of 5-fluorouracil leads to increased potency, resulting in improved shrinkage of solid tumors, enhanced life expectancy, and reduced side effects. This novel design and development make this system ideal for orally administrable drugs with low solubility or permeability or both and even making them effective at a lower dose.

Exploration of hybrid nanocarbon composite with polylactic acid for packaging applications.

Polylactic acid (PLA) nanocomposite films were fabricated with graphene oxide (GO) and single-walled carbon nanotubes (CNT) as a hybrid-co-filler with GOCNT fraction varying from 0.05 to 0.4% by weight. The effect of the GOCNT on the physical, thermal, morphological, gas permeation, and optical properties was investigated. The X-ray diffraction test reveals no restacking and coagulation of GOCNT in the composite films. Differential Scanning Calorimetry analysis shows an insignificant shift of glass transition and melting temperature but enhanced crystallization resulting from the existence of GOCNT as a nucleating agent. Scanning Electron Microscope scans indicate GOCNT embedded homogeneously without considerable aggregates in the PLA. Transmission of ultraviolet-visible radiation decreases to 30% with increasing fraction of GOCNT while Oxygen Transmission Rate diminishes to 67% in the film. These are attributed to the tortuous pathways provided by the well-dispersed hybrid GOCNT in the PLA. Compared to the pristine PLA film, the composite film shows an increase of 75% and 130% in the tensile strength and Young's modulus, respectively. Taken together, all of these improvements observed in the hybrid GOCNT-PLA composites should provide useful guidelines in customizing designs for applications across a range of fields including packaging, life sciences, cosmetics, and conventional synthetic plastics.

Toward a self-deploying shape memory polymer neuronal electrode.

The widespread application of neuronal probes for chronic recording of brain activity and functional stimulation has been slow to develop partially due to long-term biocompatibility problems with existing metallic and ceramic probes and the tissue damage caused during probe insertion. Stiff probes are easily inserted into soft brain tissue but cause astrocytic scars that become insulating sheaths between electrodes and neurons. In this communication, we explore the feasibility of a new approach to the composition and implantation of chronic electrode arrays. We demonstrate that softer polymer-based probes can be inserted into the olfactory bulb of a mouse and that slow insertion of the probes reduces astrocytic scarring. We further present the development of a micromachined shape memory polymer probe, which provides a vehicle to self-deploy an electrode at suitably slow rates and which can provide sufficient force to penetrate the brain. The deployment rate and composition of shape memory polymer probes can be tailored by polymer chemistry and actuator design. We conclude that it is feasible to fabricate shape memory polymer-based electrodes that would slowly self-implant compliant conductors into the brain, and both decrease initial trauma resulting from implantation and enhance long-term biocompatibility for long-term neuronal measurement and stimulation.

A classifier based on the artificial neural network approach for cardiologic auscultation in pediatrics.

OBJECTIVE: This research work was aimed at developing a reliable screening device for diagnosis of heart murmurs in pediatrics. This is a significant problem in pediatric cardiology because of the high rate of incidence of heart murmurs in this population (reportedly 77-95%), of which only a small fraction arises from congenital heart disease. The screening devices currently available (e.g. chest X-ray, electrocardiogram, etc.) suffer from poor sensitivity and specificity in detecting congenital heart disease. Thus, patients with heart murmurs today are frequently assessed by consultation as well with advanced imaging techniques. The most prominent among these is echocardiography. However, echocardiography is expensive and is usually only available in healthcare centers in major cities. Thus, for patients being evaluated with a heart murmur, developing a more accurate screening device is vital to efforts in reducing health care costs. METHODS AND MATERIAL: The data set was collected from incoming pediatrics at the cardiology clinic of The Children's Hospital (Denver, Colorado), on whom echocardiography had been performed to identify congenital heart disease. Recordings of approximately 10-15s duration were made at 44,100Hz and the average record length was approximately 60,000 points. The best three cycles with respect to signal quality sounds were extracted from the original recording. The resulting data comprised 241 examples, of which 88 were examples of innocent murmurs and 153 were examples of pathological murmurs. The selected phonocardiograms were subject to the digital signal processing (DSP) technique of fast Fourier transform (FFT) to extract the energy spectrum in frequency domain. The spectral range was 0-300Hz at a resolution of 1Hz. The processed signals were used to develop statistical classifiers and a classifier based on our in-house artificial neural network (ANN) software. For the latter, we also tried enhancements to the basic ANN scheme. These included a method for setting the decision-threshold and a scheme for consensus-based decision by a committee of experts. RESULTS: Of the different classifiers tested, the ANN-based classifier performed the best. With this classifier, we were able to achieve classification accuracy of 83% sensitivity and 90% specificity in discriminating between innocent and pathological heart murmurs. For the problem of discrimination between innocent murmurs and murmurs of the ventricular septal defect (VSD), the accuracy was higher, with sensitivity of 90% and specificity of 93%. CONCLUSIONS: An ANN-based approach for detection and identification of congenital heart disease in pediatrics from heart murmurs can result in an accurate screening device. Considering that only a simple feature set was used for classification, the results are very encouraging and point out the need for further development using improved feature set with more potent diagnostic variables.

The Feasibility of a Smart Surgical Probe for Verification of IRE Treatments Using Electrical Impedance Spectroscopy.

SIGNIFICANCE: Irreversible electroporation (IRE) is gaining popularity as a focal ablation modality for the treatment of unresectable tumors. One clinical limitation of IRE is the absence of methods for real-time treatment evaluation, namely actively monitoring the dimensions of the induced lesion. This information is critical to ensure a complete treatment and minimize collateral damage to the surrounding healthy tissue. GOAL: In this study, we are taking advantage of the biophysical properties of living tissues to address this critical demand. METHODS: Using advanced microfabrication techniques, we have developed an electrical impedance microsensor to collect impedance data along the length of a bipolar IRE probe for treatment verification. For probe characterization and interpretation of the readings, we used potato tuber, which is a suitable platform for IRE experiments without having the complexities of in vivo or ex vivo models. We used the impedance spectra, along with an electrical model of the tissue, to obtain critical parameters such as the conductivity of the tissue before, during, and after completion of treatment. To validate our results, we used a finite element model to simulate the electric field distribution during treatments in each potato. RESULTS: It is shown that electrical impedance spectroscopy could be used as a technique for treatment verification, and when combined with appropriate FEM modeling can determine the lesion dimensions. CONCLUSIONS: This technique has the potential to be readily translated for use with other ablation modalities already being used in clinical settings for the treatment of malignancies.

Fast response temperature measurement and highly reproducible heating methods for 96-well plates.

Hyperthermia, the procedure of exposing cells to a temperature between 42 degrees and 49 degrees C, has been shown to be a promising approach for cancer treatment. To understand the underlying mechanisms of hyperthermic killing of cancer cells, it is critical to have an accurate temperature measurement technique and a heating method with high reproducibility. To this end, we have developed a method using fine thermocouples with fast response time to measure the temperatures in multiple wells of a 96-well plate. The accuracy of temperature measurement was +/- 0.2 degree C. Such a capability allows a complete record of the time and temperature of the treatment procedure and helps define an accurate thermal dose. We have also compared several methods for heating 96-well plates and found that use of copper blocks in contact with the lower surface of the 96-well plate in an incubator provides a highly reproducible heating method. The common method of using water bath to heat cells in vitro resulted in a decrease of cell viability even at the control temperature of 37 degrees C and a decrease in the reproducibility of certain biological assays. In summary, using these improved techniques, proposed thermal dose can be defined more precisely, and highly reproducible heating in vitro can be achieved.

Studying the activity of antitubercluosis drugs inside electrospun polyvinyl alcohol, polyethylene oxide, and polycaprolacton nanofibers.

The activity of antituberculosis drugs (streptomycin sulfate, isoniazid, pyrazinamid, and clarithromycin) embedded in biodegradable nanofibers against Mycobacterium avium has been studied by broth dilution assay and by agar plate assay. These drugs have also been embedded in electrospun polyvinyl alcohol (PVA), polyethylene oxide (PEO), and polycaprolacton (PCL) nanofibers to design a new single tablet containing first-line antituberculosis drugs. Our results show that antituberculosis drugs are active at tiny amounts (up to 300 µg mL(-1) of solvent). However, within polymer matrices, high amounts of drugs are required to avoid unwanted weak interactions within PEO and PCL matrices. The successful design of a single tablet containing required amounts of antituberculosis drugs is essential for the full treatment of tuberculosis in patients with HIV.

Differential and directional effects of perfusion on electrical and thermal conductivities in liver.

Two different measurement probes--an electrical probe and a thermal conductivity probe--were designed, fabricated, calibrated, and used in experimental studies on a pig liver model that was designed to control perfusion rates. These probes were fabricated by photolithography and mounted in 1.5-mm diameter catheters. We measured the local impedance and thermal conductivity, respectively, of the artificially perfused liver at different flow rates and, by rotating the probes, in different directions. The results show that both the local electrical conductivity and the thermal conductivity varied location to location, that thermal conductivity increased with decreased distance to large blood vessels, and that significant directional differences exist in both electrical and thermal conductivities. Measurements at different perfusion rates demonstrated that both the local electrical and local thermal conductivities increased linearly with the square root of perfusion rate. These correlations may be of great value to many energy-based biomedical applications.

Micromachined hot-wire thermal conductivity probe for biomedical applications.

This paper presents the design, fabrication, numerical simulation, and experimental validation of a micromachined probe that measures thermal conductivity of biological tissues. The probe consists of a pair of resistive line heating elements and resistance temperature detector sensors, which were fabricated by using planar photolithography on a glass substrate. The numerical analysis revealed that the thermal conductivity and diffusivity can be determined by the temperature response induced by the uniform heat flux in the heating elements. After calibrating the probe using a material (agar gel) of known thermal conductivity, the probe was deployed to calculate the thermal conductivity of Crisco. The measured value is in agreement with that determined by the macro-hot-wire probe method to within 3%. Finally, the micro thermal probe was used to investigate the change of thermal conductivity of pig liver before and after RF ablation treatment. The results show an increase in thermal conductivity of liver after the RF ablation.

A uniaxial bioMEMS device for imaging single cell response during quantitative force-displacement measurements.

A microfabricated device has been developed for imaging of a single, adherent cell while quantifying force under an applied displacement. The device works in a fashion similar to that of a displacement-controlled uniaxial tensile machine. The device was calibrated using a tipless atomic force microscope (AFM) cantilever and shows excellent agreement with the calculated spring constant. A step input was applied to a single, adherent fibroblast cell and the viscoelastic response was characterized with a mechanical model. The adherent fibroblast was imaged by use of epifluorescence and phase contrast techniques.

Biocompatibility of atomic layer-deposited alumina thin films.

Presented in this paper is a study of the biocompatibility of an atomic layer-deposited (ALD) alumina (Al2O3) thin film and an ALD hydrophobic coating on standard glass cover slips. The pure ALD alumina coating exhibited a water contact angle of 55 degrees +/- 5 degrees attributed, in part, to a high concentration of -OH groups on the surface. In contrast, the hydrophobic coating (tridecafluoro-1,1,2,2-tetrahydro-octyl-methyl-bis(dimethylamino)silane) had a water contact angle of 108 degrees +/- 2 degrees. Observations using differential interference contrast microscopy on human coronary artery smooth muscle cells showed normal cell proliferation on both the ALD alumina and hydrophobic coatings when compared to cells grown on control substrates. These observations suggested good biocompatibility over a period of 7 days in vitro. Using a colorimetric assay technique to assess cell viability, the cellular response between the three substrates can be differentiated to show that the ALD alumina coating is more biocompatible and that the hydrophobic coating is less biocompatible when compared to the control. These results suggest that patterning a substrate with hydrophilic and hydrophobic groups can control cell growth. This patterning can further enhance the known advantages of ALD alumina, such as conformality and excellent dielectric properties for bio-micro electro mechanical systems (Bio-MEMS) in sensors, actuators, and microfluidics devices.

Hyperthermia induces endoplasmic reticulum-mediated apoptosis in melanoma and non-melanoma skin cancer cells.

Hyperthermia has been revived as a promising approach for cancer treatment. To understand the underlying mechanisms of hyperthermic killing of cancer cells, we examined the cytotoxic effects of hyperthermia on various skin cancer cell lines using cell viability, morphological analyses, and caspase activation assays. Hyperthermia induced cytotoxicity in a time- and temperature-dependent manner. At middle dose/time combinations, heat-induced apoptosis, whereas at higher doses, necrosis was the mechanism of cell death. To investigate the mechanisms of hyperthermia-induced apoptosis, we examined the activation of extrinsic (Caspase 8) and intrinsic (Caspase 9) apoptotic pathways. Hyperthermia did not activate Caspases 8 or 9, but did activate Caspase 3/7, suggesting a non-conventional apoptotic pathway. Last, analysis of Grp78 expression and Caspase 12 or 4 activation indicated that hyperthermia induced endoplasmic reticulum-mediated apoptosis. Thus, hyperthermia induced apoptosis in two types of skin cancer cells through endoplasmic reticulum-mediated apoptosis and not through the classical intrinsic or extrinsic apoptosis pathways. Hyperthermia may be a promising treatment for basal cell carcinoma and melanoma, bypassing the antiapoptotic defenses concentrated in the intrinsic and extrinsic apoptosis pathways. These results also raise the possibility that heat may be combined with other approaches for induction of apoptosis to achieve synergistic killing of skin cancers.

Foam-based optical absorber for high-power laser radiometry.

We report damage threshold measurements of novel absorbers comprised of either liquid-cooled silicon carbide or vitreous carbon foams. The measurements demonstrate damage thresholds up to 1.6x10(4) W/cm(2) at an incident circular spot size of 2 mm with an absorbance of 96% at 1.064 microm. We present a summary of the damage threshold as a function of the water flow velocity and the absorbance measurements. We also present a qualitative description of a damage mechanism based on a two-phase heat transfer between the foam and the flowing water.

A uniaxial bioMEMS device for quantitative force-displacement measurements.

There is a need for experimental techniques that allow the simultaneous imaging of cellular cystoskeletal components with quantitative force measurements on single cells. A bioMEMS device has been developed for the application of strain to a single cell while simultaneously quantifying its force response. The prototype device presented here allows the mechanical study of a single, adherent cell in vitro. The device works in a fashion similar to a displacement-controlled uniaxial tensile machine. The device is calibrated using an AFM cantilever and shows excellent agreement with the calculated spring constant. The device is demonstrated on a single fibroblast. The force response of the cell is seen to be linear until the onset of de-adhesion with the de-adhesion from the cell platform occurring at a force of approximately 1500 nN.