Characteristics of Artemether-Loaded Poly(lactic-co-glycolic) Acid Microparticles Fabricated by Coaxial Electrospray: Validation of Enhanced Encapsulation Efficiency and Bioavailability.

Artemether is one of the most effective drugs for the treatment of chloroquine-resistant and Plasmodium falciparum strains of malaria. However, its therapeutic potency is hindered by its poor bioavailability. To overcome this limitation, we have encapsulated artemether in poly(lactic-co-glycolic) acid (PLGA) core-shell microparticles (MPs) using the coaxial electrospray method. With optimized process parameters including liquid flow rates and applied electric voltages, experiments are systematically carried out to generate a stable cone-jet mode to produce artemether-loaded PLGA-MPs with an average size of 2 µm, an encapsulation efficiency of 78 ± 5.6%, and a loading efficiency of 11.7%. The in vitro release study demonstrates the sustained release of artemether from the core-shell structure in comparison with that of plain artemether and that of MPs produced by single-axial electrospray without any relevant cytotoxicity. The in vivo studies are performed to evaluate the pharmacokinetic characteristics of the artemether-loaded PLGA-MPs. Our study implies that artemether can be effectively encapsulated in a protective shell of PLGA for controlled release kinetics and enhanced oral bioavailability.

Three-dimensional simulation of mass transfer in artificial kidneys.

In this work, the three-dimensional velocity and concentration fields on both the blood and dialysate sides in an artificial kidney were simulated, taking into account the effects of the flow profiles induced by the inlet and outlet geometrical structures and the interaction between the flows of blood and dialysate. First, magnetic resonance imaging experiments were performed to validate the mathematical model. Second, the effects of the flow profiles induced by the blood and dialysate inlet and outlet geometrical structures on mass transfer were theoretically investigated. Third, the clearance of toxins was compared with the clearance value calculated by a simple model that is based on the ideal flow profiles on both the blood and dialysate sides. Our results show that as the blood flow rate increases, the flow field on the blood side becomes less uniform; however, as the dialysate flow rate increases, the flow field on the dialysate side becomes more uniform. The effect of the inlet and outlet geometrical structures of the dialysate side on the velocity and concentration fields is more significant than that of the blood side. Due to the effects of the flow profiles induced by the inlet and outlet geometrical structures, the true clearance of toxins is lower than the ideal clearance, especially when the dialysate flow rate is low or the blood flow rate is high. The results from this work are significant for the structural optimization of artificial kidneys and the accurate prediction of toxin clearance.

A new method to increase the adsorption of protein-bound toxins in artificial liver support systems.

In this work, a new method, called the preconcentration method (PCM), is proposed to increase the adsorption of protein-bound toxins onto adsorbents in artificial liver support systems. In the PCM, a concentrator is installed before the inlet of the adsorbent cartridge. This method is validated in an experiment using activated carbon to remove albumin-bound bilirubin, and the mechanism of the increase in adsorption is theoretically explained with breakthrough curve and equilibrium adsorption analyses. Our results show that when this PCM is used, the mass transfer rate of bilirubin from solution to activated carbon is enhanced, the adsorbed bilirubin amount per unit mass of activated carbon is greatly increased, and more albumin-bound bilirubin molecules are quickly removed from the albumin solution. When the concentration ratio (the ratio of the inlet flow rate to the outflow rate of the concentrator) is 2.59, the adsorption efficiency of activated carbon at 120 min is increased by approximately 36%. Only approximately 60 min is required for the bilirubin concentration to decrease from 19.3 to 13.0 mg/dL; however, without the PCM, nearly 180 min is needed. In addition, by adjusting the concentration ratio, the adsorption of albumin-bound bilirubin onto activated carbon can be further increased.

Microfabricated thermal conductivity sensor: a high resolution tool for quantitative thermal property measurement of biomaterials and solutions.

Obtaining accurate thermal properties of biomaterials plays an important role in the field of cryobiology. Currently, thermal needle, which is constructed by enclosing a manually winded thin metal wire with an insulation coating in a metallic sheath, is the only available device that is capable of measuring thermal conductivity of biomaterials. Major drawbacks, such as macroscale sensor size, lack of versatile format to accommodate samples with various shapes and sizes, neglected effects of heat transfer inside the probe and thermal contact resistance between the sensing element and the probe body, difficult to mass produce, poor data repeatability and reliability and labor-intense sensor calibration, have significantly reduced their potential to be an essential measurement tool to provide key thermal property information of biological specimens. In this study, we describe the development of an approach to measure thermal conductivity of liquids and soft bio-tissues using a proof-of-concept MEMS based thermal probe. By employing a microfabricated closely-packed gold wire to function as the heater and the thermistor, the presented thermal sensor can be used to measure thermal conductivities of fluids and natural soft biomaterials (particularly, the sensor may be directly inserted into soft tissues in living animal/plant bodies or into tissues isolated from the animal/plant bodies), where other more standard approaches cannot be used. Thermal standard materials have been used to calibrate two randomly selected thermal probes at room temperature. Variation between the obtained system calibration constants is less than 10%. By incorporating the previously obtained system calibration constant, three randomly selected thermal probes have been successfully utilized to measure the thermal conductivities of various solutions and tissue samples under different temperatures. Overall, the measurements are in agreement with the recommended values (percentage error less than 5%). The microfabricated thermal conductivity sensor offers superior characteristics compared to those traditional macroscopic thermal sensors, such as, (a) reduced thermal mass and thermal resistivity, (b) improved thermal contact between sensor and sample, (c) easy to manufacture with mass production capability, (d) flexibility to reconfigure sensor geometries for measuring samples with various sizes and shapes, and (e) reduced calibration workload for all sensors microfabricated from the same batch. The MEMS based thermal conductivity sensor is a promising approach to overcome the inherent limitations of existing macroscopic devices and capable of delivering accurate thermal conductivity measurement of biomaterials with various shapes and sizes.

Size-selective immunofluorescence of Mycobacterium tuberculosis cells by capillary- and viscous forces.

Rapid, low cost screening of tuberculosis requires an effective enrichment method of Mycobacterium tuberculosis (MTB) cells. Currently, microfiltration and centrifugation steps are frequently used for sample preparation, which are cumbersome and time-consuming. In this study, the size-selective capturing mechanism of a microtip-sensor is presented to directly enrich MTB cells from a sample mixture. When a microtip is withdrawn from a spherical suspension in the radial direction, the cells that are concentrated by AC electroosmosis are selectively enriched to the tip due to capillary- and viscous forces. The size-selectivity is characterized by using polystyrene microspheres, which is then applied to size-selective capture of MTB from a sample mixture. Our approach yields a detection limit of 800 cells mL(-1), one of the highest-sensitivity immunosensors to date.

Blood-membrane interactions during dialysis.

In extracorporeal renal replacement therapies, the dialyzer is not only the site at which solute removal occurs but also the extracorporeal circuit component having the largest surface area exposed to blood. Therefore, it is not surprising that interactions between blood components and the dialyzer membrane influence the dialysis procedure in several ways. Based on engineering principles, fluid flow along a surface such as membrane results in the development of a boundary layer which can influence solute removal. Furthermore, the exposure of blood to any extracorporeal artificial surface results in the activation of several pathways within the body, including those involving coagulation and complement activation. One of the byproducts of this generalized activation process is protein adsorption to the membrane surface, another phenomenon which can have a significant impact on solute removal. In this article, a detailed review of the ways in which blood-membrane interactions influence solute removal during hemodialysis and related therapies is provided. The influences of secondary membrane formation and boundary layer/concentration polarization effects on solute removal are specifically discussed. Furthermore, the importance of adsorption as a specific removal mechanism for low-molecular weight proteins by highly permeable synthetic membranes is highlighted.

Membranes and Sorbents.

For continuous renal replacement therapy (CRRT), the extracorporeal filter provides solute depuration, fluid removal, and control of electrolyte and acid-base balance in critically ill patients with acute kidney injury (AKI). The membranes comprising CRRT filters are almost exclusively based on hollow fiber designs and, while adapted from the chronic hemodialysis field, have features that are specific to the requirements of CRRT nevertheless. In addition, these devices have evolved through the 40 years of CRRT in response to changes in clinical practice and the desire to extend the solute removal spectrum. For some critically ill patients, more targeted removal of specific compounds poorly cleared by standard CRRT can be attempted with techniques based on adsorption. Sorbent hemoperfusion is now being applied more broadly in critically ill patients, especially in those with sepsis and systemic inflammation. In this review, the manner in which CRRT membranes and extracorporeal sorbents have evolved over the past 40 years for the treatment of critically ill patients with AKI and other disorders is described.

Development of a microfluidic device for determination of cell osmotic behavior and membrane transport properties.

An understanding of cell osmotic behavior and membrane transport properties is indispensable for cryobiology research and development of cell-type-specific, optimal cryopreservation conditions. A microfluidic perfusion system is developed here to measure the kinetic changes of cell volume under various extracellular conditions, in order to determine cell osmotic behavior and membrane transport properties. The system is fabricated using soft lithography and is comprised of microfluidic channels and a perfusion chamber for trapping cells. During experiments, rat basophilic leukemia (RBL-1 line) cells were injected into the inlet of the device, allowed to flow downstream, and were trapped within a perfusion chamber. The fluid continues to flow to the outlet due to suction produced by a Hamilton Syringe. Two sets of experiments have been performed: the cells were perfused by (1) hypertonic solutions with different concentrations of non-permeating solutes and (2) solutions containing a permeating cryoprotective agent (CPA), dimethylsulfoxide (Me(2)SO), plus non-permeating solute (sodium chloride (NaCl)), respectively. From experiment (1), cell osmotically inactive volume (V(b)) and the permeability coefficient of water (L(p)) for RBL cells are determined to be 41% [n=18, correlation coefficient (r(2)) of 0.903] of original/isotonic volume, and 0.32+/-0.05 microm/min/atm (n=8, r(2)>0.963), respectively, for room temperature (22 degrees C). From experiment (2), the permeability coefficient of water (L(p)) and of Me(2)SO (P(s)) for RBL cells are 0.38+/-0.09 microm/min/atm and (0.49+/-0.13) x 10(-3)cm/min (n=5, r(2)>0.86), respectively. We conclude that this device enables us to: (1) readily monitor the changes of extracellular conditions by perfusing single or a group of cells with prepared media; (2) confine cells (or a cell) within a monolayer chamber, which prevents imaging ambiguity, such as cells overlapping or moving out of the focus plane; (3) study individual cell osmotic response and determine cell membrane transport properties; and (4) reduce labor requirements for its disposability and ensure low manufacturing costs.

Solute Transport in Hemodialysis: Advances and Limitations of Current Membrane Technology.

Extracorporeal therapy for end-stage renal disease is now provided to more than three million patients globally. Nearly all treatments are performed with filters containing hollow fiber membranes, removing solutes and water by diffusion, convection, and ultrafiltration. In this review, we will provide a detailed quantitative analysis of the transport processes involved in different hemodialysis (HD) therapies. We will also report some technical aspects of hollow fiber membranes and filters composed of them along with the mechanisms of solute and water removal for such devices. Diffusive mass transfer will be assessed according to the three major aspects of a hollow fiber filter (blood compartment, membrane, and dialysate compartment). With regard to convective transport, the importance of internal filtration as a solute removal mechanism in high-flux HD will be highlighted, along with the critical role that blood/membrane interactions assume in filtration-based therapies.

Sensing and Sensibility: Single-Islet-based Quality Control Assay of Cryopreserved Pancreatic Islets with Functionalized Hydrogel Microcapsules.

Despite decades of research and clinical studies of islet transplantations, finding simple yet reliable islet quality assays that correlate accurately with in vivo potency is still a major challenge, especially for real-time and single-islet-based quality assessment. Herein, proof-of-concept studies of a cryopreserved microcapsule-based quality control assays are presented for single islets. Individual rat pancreatic islets and fluorescent oxygen-sensitive dye (FOSD) are encapsulated in alginate hydrogel microcapsules via a microfluidic device. To test the susceptibility of the microcapsules and the FOSD to cryopreservation, the islet microcapsules containing FOSD are cryopreserved and the islet functionalities (adenosine triphosphate, static insulin release measurement, and oxygen consumption rate) are assessed after freezing and thawing steps. The cryopreserved islet capsules with FOSD remain functional after encapsulation and freezing/thawing procedures, validating a simple yet reliable individual-islet-based quality control method for the entire islet processing procedure prior to transplantation. This work also demonstrates that the functionality of cryopreserved islets can be improved by introducing trehalose into the routinely used cryoprotectant dimethyl sulfoxide. The functionalized alginate hydrogel microcapsules with embedded FOSD and optimized cryopreservation protocol presented in this work serve as a versatile islet quality assay and offer tremendous promise for tackling existing challenges in islet transplantation procedures.

[Experiment on the formation and characterizations of anodic alumina membrane for use in hemodialysis].

The correlations between various formation conditions and the membrane pore characterizations of the anodic alumina membrane were investigated for seeking the optimal conditions for the formation of anodic alumina membrane. High purity aluminum foils were used as the starting materials. The anodizations were conducted under three types of electrolytes, 3% sulfuric acid, 5% sulfuric acid and 2.7% oxalic acid, respectively, with different voltages at 0 degrees C for 48 hours. The characterizations of the pore size, the effective porosity and the pore porosity were observed and determined by scanning electron microscopy. The hydraulic conductance of the membranes was measured to confirm that the pores were open and to evaluate the permselectivity of the membranes. The experimental results showed that the ordered pore arrays were obtained for oxidation under our experimental conditions. While the forming voltage was increasing, the pore size and pore porosity increased significantly (P < 0.05), and the effective porosity decreased significantly (P < 0.05). The pore size formed with 3% sulfuric acid or 5% sulfuric acid was much smaller than that with 2.7% oxalic acid as an electrolyte. The hydraulic conductance of anodic alumina membrane that formed under our experimental condition was high than those of the membranes currently available in clinical procedures. The results suggested that the optimal conditions for the formation of anodic alumina membrane to be used in hemodialysis should be 3% or 5% sulfuric acid with 12.5 V to 17.5 V at 0 degrees C for 48 hours.

Evaluation of nano-porous alumina membranes for hemodialysis application.

Globally, kidney failure has consistently been a major health problem. The number of patients suffering from kidney failure is radically increasing. Some studies forecast an exponential growth in the number of kidney failure patients during the coming years. This emphasizes the importance of hemodialysis (HD) membranes. Current dialysis membranes (cellulose based and synthetic polymer membranes) have irregular pore shapes and sizes, nonuniform pore distribution and limited reusable capability, which leads to low efficiency of toxin removal. New alumina membranes with uniform, controllable and well-structured nanoscale pores, channeled pores aligned perpendicular to the membrane plane, high porosity, high thermal and chemical resistance, and better mechanical properties are certainly preferable to currently used membranes. Determination of transport properties of alumina membranes will assist in the development of the alumina membranes for enhancing hemodialysis. Experiments were performed to evaluate hydraulic permeability, solute diffusive permeability, sieving coefficient, and clearance of four solutes (urea, creatinine, Vancomycin, and inulin) for alumina membrane. Based on comparison of these values against those of polyethersulfone (PES) membranes, transport performance of alumina membrane was determined. Hydraulic conductivity of the alumina membrane was approximately twice that of the PES membrane and inulin sieving coefficient for alumina membrane is approximately 21% higher than that for PES membrane. Alumina membrane has higher solute clearances and no albumin leakage, which makes it an effective replacement for current dialysis membranes.

Development of a single mode electromagnetic resonant cavity for rewarming of cryopreserved biomaterials.

An electromagnetic (EM) heating system is developed to achieve the rapid and uniform warming of cryopreserved biomaterials. Using the heating system, a rectangular resonant cavity is excited in TE101 mode at frequencies near 434 MHz. In experiments, a spherical phantom of biomaterial with a diameter of 36 mm is placed at the center of the cavity. The phantom is first cooled down to about -80 degrees C within the cavity and then thawed by EM absorption. Results show that EM warming can produce much higher warming rate than conventional water-bath warming method. The spatial temperature distribution in the phantom during EM warming is also more uniform than that during the water-bath warming.

Properties of membranes used for hemodialysis therapy.

Recent trends show a progressive increase in the use of modified cellulosic and synthetic dialyzers and a corresponding decrease in the utilization rate of unmodified cellulosic dialyzers. The purpose of this article is to describe current membrane and dialyzer technology, with the focus almost entirely on modified cellulosic and synthetic membranes. A general overview of membrane-related determinants of dialyzer performance is first presented, followed by a discussion of specific characteristics of some of the more commonly used membranes and dialyzers.

A numerical and experimental study of mass transfer in the artificial kidney.

To develop a more efficient and optimal artificial kidney, many experimental approaches have been used to study mass transfer inside, outside, and cross hollow fiber membranes with different kinds of membranes, solutes, and flow rates as parameters. However, these experimental approaches are expensive and time consuming. Numerical calculation and computer simulation is an effective way to study mass transfer in the artificial kidney, which can save substantial time and reduce experimental cost. This paper presents a new model to simulate mass transfer in artificial kidney by coupling together shell-side, lumen-side, and transmembrane flows. Darcy's equations were employed to simulate shell-side flow, Navier-Stokes equations were employed to simulate lumen-side flow, and Kedem-Katchalsky equations were used to compute transmembrane flow. Numerical results agreed well with experimental results within 10% error. Numerical results showed the nonuniform distribution of flow and solute concentration in shell-side flow due to the entry/exit effect and Darcy permeability. In the shell side, the axial velocity in the periphery is higher than that in the center. This numerical model presented a clear insight view of mass transfer in an artificial kidney and may be used to help design an optimal artificial kidney and its operation conditions to improve hemodialysis.

Effect of flow baffles on the dialysate flow distribution of hollow-fiber hemodialyzers: a nonintrusive experimental study using MRI.

We used an innovative, nonintrusive MRI technique called the two-dimensional (2D) Phase-Contrast (2DPC) velocity-imaging technique to investigate the effect of flow baffles on the dialysate-side flow distribution in two different hollow-fiber hemodialyzers (A and B); each with flow rates between 200 and 1000 mL/min (3.33 x 10(-6) and 1.67 x 10(-5) m3/s). Our experimental results show that (1) the dialysate-side flow distribution was nonuniform with channeling flow occurred at the peripheral cross section of these hollow-fiber hemodialyzers, and (2) the existing designs of flow baffles failed to promote uniform dialysate-side flow distribution for all flow rates studies.

Improving the blood compatibility of ion-selective electrodes by employing poly(MPC-co-BMA), a copolymer containing phosphorylcholine, as a membrane coating.

The hydrogelpoly(2-methacryloyloxyethylphosphorylcholine-co-butyl methacrylate), or poly(MPC-co-BMA), was used as a coating for polyurethane- and poly(vinyl chloride)-based membranes to develop ion-selective electrodes (ISEs) with enhanced blood compatibility. Adverse interactions of poly(MPC-co-BMA) with blood were diminished due to the phosphorylcholine functionalities of the hydrogel, which mimic the phospholipid polar groups present on the surface of many cell membranes. As demonstrated by immunostaining, hydrogel-coated PVC membranes soaked in platelet-rich plasma showed less adhesion and activation of platelets than uncoated PVC membranes, indicating an improvement in biocompatibility owing to the hydrogel. Furthermore, little differences in the potentiometric response characteristics, e.g., slope, detection limit, and selectivity, of ISEs employing uncoated and coated membranes were observed.