Effects of layer-by-layer coating on activated carbon electrodes for capacitive deionization.

Recently, the need for obtaining, reusing, or purifying water has become a crucial issue. The capacitive deionization (CDI) method, which is based on the electric double layer (EDL) concept, can be applied to ion adsorption from an aqueous solution. This process is carried out by applying a potential difference to highly porous electrodes while pumping salty solution between them, partially removing the ions present in the solution and keeping them in the surface of the electrodes. The use of coated carbon electrodes with one polyelectrolyte layer, turning them into "soft electrodes" (SEs), has been proved to improve the efficiency of the system with respect to its original configuration. In this work, we investigate the effect on the ion adsorption and the efficiency of the process when implementing the coating technique known as layer-by-layer (LbL) on the electrode. This consists in successively coating the electrode surfaces with polyelectrolyte layers, alternating their charge polarity in each step. We tested the effect of the number of layers deposited, as well as the impact of this technique by using different carbons. We found that the second polyelectrolyte layer adheres more than the first layer, serving as a support or seed when it is not dense and uniformly distributed. In contrast, if the first layer is well adhered, a third layer is needed to observe improvements in adsorption and process efficiency. The adsorption of the polymer layers depends in any instance on the porosity of the carbon.

First steps in the formulation of praziquantel nanosuspensions for pharmaceutical applications.

Praziquantel (PZQ), a broad spectrum anthelmintic drug, cannot be found in acceptable dosage forms for elderly patients, paediatric patients, and for veterinary use. In fact, very little has been done up to now in the formulation of liquid dosage forms, being they always formulated for parenteral administration. To beat this important challenge, it was accomplished a comprehensive analysis of the influence of two elementary physicochemical aspects, i.e. surface thermodynamic and electrokinetic properties, on the colloidal stability of PZQ nanosuspensions. The hydrophobic character of the drug, intensely determining the flocculation curves, was confirmed by the thermodynamic characterization. The electrophoretic characterization, in combination with the sedimentation and relative absorbance versus time curves, highlighted that the electrical double layer thickness and the surface charge can play an essential role in the stability of the pharmaceutical colloid. Finally, it was demonstrated that controlling the pH values and the incorporation of electrolytes can help in formulating PZQ aqueous nanosuspensions with appropriate stability and redispersibility behaviours for pharmaceutical use.

Electro-Orientation of Silver Nanowires in Alternating Fields.

In this work, we analyze the orientation of silver nanowires immersed in aqueous solutions, under the effect of alternating electric fields in a broad frequency range covering from a few Hz to several MHz. The degree of orientation is experimentally determined by electro-optical techniques, which present the advantage of measuring multiple particles at the same time. In the electro-orientation spectrum, we observe frequency dispersion in the kHz range and provide a theoretical explanation for this behavior: at high frequencies, charge separation in the nanoparticles leads to a large induced dipole responsible for strong orientation. On the other hand, at low frequencies, redistribution of the ions in solution gives rise to an induced double layer that screens the dipolar fields, and as a consequence, the degree of orientation decreases. Moreover, we measure the transient response when the electric field is switched off, from which the size distribution of the polydisperse sample is obtained. The results match those given by electron microscopy determinations.

Charging Poly(methyl Methacrylate) Latexes in Nonpolar Solvents: Effect of Particle Concentration.

The electrophoresis of a well-established model system of charged colloids in nonpolar solvents has been studied as a function of particle volume fraction at constant surfactant concentration. Dispersions of poly(12-hydroxystearic acid)-stabilized poly(methyl methacrylate) (PMMA) latexes in dodecane were prepared with added Aerosol OT surfactant as the charging agent. The electrophoretic mobility (µ) of the PMMA latexes is found to decrease with particle concentration. The particles are charged by a small molecule charging agent (AOT) at finite concentration, and this makes the origin of this decrease in µ unclear. There are two suggested explanations. The decrease could either be due to the reservoir of available surfactant being exhausted at high particle concentrations or the interactions between the charged particles at high particle number concentrations. Contrast-variation small-angle neutron scattering measurements of PMMA latexes and deuterated AOT-d34 surfactant in latex core contrast-matched solvent were used to study the former, and electrokinetic modeling was used to study the latter. As the same amount of AOT-d34 is found to be incorporated with the latexes at all volume fractions, the solvodynamic and electrical interactions between particles are determined to be the explanation for the decrease in mobility. These measurements show that, for small latexes, there are interactions between the charged particles at all accessible particle volume fractions and that it is necessary to account for this to accurately determine the electrokinetic ζ potential.

Use of Soft Electrodes in Capacitive Deionization of Solutions.

All efforts to obtain, reuse or purify water are extremely significant for society. Recently, researchers have begun to delve in an idea born decades ago: the desalination of water using highly porous electrodes. It is based on a fundamental aspect of electrical double layers, namely, their huge capacitance. The ions of a solution can be partially removed under the application of an electric field when the solution fills the space between porous electrodes, either bare (CDI, or capacitive deionization), coated with ionic exchange membranes (MCDI) or chemically treated (inverted-CDI or i-CDI). One of the challenges of the last years was to explore new materials and arrangements to improve the efficiency of the system. In this work, we propose a new approach inspired in the electrokinetics of soft particles: a layer of polyelectrolyte (cationic on one electrode, anionic on the opposite one) coats the carbon electrodes, converting them in a sort of "soft" electrode pair. We present a theoretical model and a set of experiments showing how soft electrodes can be successfully employed in capacitive deionization.

Development and Characterization of Magnetite/Poly(butylcyanoacrylate) Nanoparticles for Magnetic Targeted Delivery of Cancer Drugs.

A great attention is presently paid to the design of drug delivery vehicles based on surface-modified magnetic nanoparticles. They can, in principle, be directed to a desired target area for releasing their drug payload, a process triggered by pH, temperature, radiation, or even magnetic field. To this, the possibility of forming part of diagnostic tools by enhanced magnetic resonance imaging or that of further treatment by magnetic hyperthermia can be added. Bare particles are rapidly eliminated from the bloodstream by the phagocyte mononuclear system, leading to short biological half-life. It is hence required to coat them in order to increase their biocompatibility and facilitate the drug incorporation. In this work, magnetite nanoparticles were coated with poly(butylcyanoacrylate) (PBCA) manufactured and characterized with regard to their physical properties and their suitability as a platform for magnetically controlled drug delivery. The average diameter of magnetite and core-shell nanoparticles was 97 ± 19 and 140 ± 20 nm, respectively. Infrared analysis, electrophoretic mobility, surface thermodynamics analysis, and X-ray diffraction all confirmed that the magnetic particles were sufficiently covered by the polymer in the composite nanoparticles. In addition, assays using normal (CCD-18 and MCF-10A) and tumoral (T-84 and MCF-7) cell lines derived from colon and breast tissue, respectively, demonstrated that nanocomposites have low or negligible cytotoxicity. It is concluded that PBCA-coated magnetite core-shell nanoparticles represent a remarkable promise as a platform for magnetically controlled drug delivery.

Electric birefringence spectroscopy of montmorillonite particles.

Electric birefringence (EB) of suspensions of anisotropic particles can be considered an electrokinetic phenomenon in a wide sense, as both liquid motions and polarization of the electrical double layer (EDL) of the particles participate in the process of particle orientation under the applied field. The EB spectrum can be exploited for obtaining information on the dimensions, average value and anisotropy of the surface conductivity of the particles, and the concentration and Maxwell-Wagner polarization of the EDLs. It is thus a highly informative technique, applicable to non-spherical particles. In this paper, we investigate the birefringent response of plate-like montmorillonite particles as a function of the frequency and amplitude of the applied AC electric field, for different compositions (pH, ionic strength, particle concentration) of the suspensions. The transient electric birefringence (i.e., the decay of the refractive index anisotropy with time when the field is switched off) is used for estimating the average dimensions of the particle axes, by modeling it as an oblate spheroid. The obtained values are very similar to those deduced from electron microscopy determinations. The frequency spectra show a very distinct behaviour at low (on the order of a few Hz) and high (up to several MHz) frequencies: the α and Maxwell-Wagner-O'Konski relaxations, characteristic of EDLs, are detected at frequencies above 10 kHz, and they can be well explained using electrokinetic models for the polarization of EDLs. At low frequencies, in contrast, the birefringence changes to negative, an anomalous response meaning that the particles tend to orient with their symmetry axis parallel to the field. This anomaly is weaker at basic pH values, high ionic strengths and low concentrations. The results can be explained by considering the polydispersity of real samples: the fastest particles redistribute around the slowest ones, inducing a hydrodynamic torque opposite to that of the field, in close similarity with the results previously described for the mixtures of anisometric particles with small amounts of spherical nanoparticles.

Electric Permittivity and Dynamic Mobility of Dilute Suspensions of Platelike Gibbsite Particles.

In this work we discuss the electrokinetic evaluation of model platelike particles. By model particles we mean homogeneous and controlled size and shape. The electrokinetic analysis in such complex geometries cannot be limited to a single data point as in usual electrophoresis in constant (dc) fields. The information can be made much richer if alternating (ac) fields with a sufficiently wide range of frequencies are used. In this case, two techniques can be applied: one is the determination of the frequency spectrum of the electric permittivity or dielectric constant (low-frequency dielectric dispersion), and the other is the electroacoustics of suspensions and the determination of the frequency dispersion of the electrophoretic mobility (dynamic or ac mobility). In this work, these techniques are used with planar gibbsite (γ-Al(OH)3) particles, modeled as oblate spheroids with a small aspect ratio. As in other laminar minerals, a particular charge distribution, differing between edges and faces, gives rise to very peculiar electrokinetic behavior. It is found that pH 7 approximately separates two distinct field responses: below that pH the dielectric dispersion and dynamic mobility data are consistent with the existence of individual, highly charged platelets, with charge mainly originating on edge surfaces. At pH 4, a low-frequency relaxation is observed, which must originated from larger particles. It is suggested that these are individual ones bridged by negatively charged fiberlike structures, coming from the partial decomposition of gibbsite particles. On the other side of the measured pH spectrum, the overall charge of the particles is low, and this probably produces aggregates with a relatively large average size, with relaxation frequencies on the low side. This is confirmed by dynamic mobility data, showing that a coherent picture of the nanostructure can be reached by combining the two techniques.

Iron/Magnetite Nanoparticles as Magnetic Delivery Systems for Antitumor Drugs.

In this study we investigate on the possible use of a new kind of magnetic nanostructures as drug delivery systems for anticancer drugs. The starting particles are formed by an inner core of iron, coated by magnetite as a stabilizing, magnetic layer. These units are further coated by a poly(ethylenglycol) (PEG) layer to make them less prone to the attack by macrophages and to favour longer stays in the blood stream. The resulting particles consist of several magnetic cores encapsulated by a polymer layer around 5 nm thick. The crystal structure of the designed nanostructures, as determined by X-ray powder diffraction, is compatible with a crystalline magnetite component, whereas the magnetization hysteresis data indicate a superparamagnetic behavior. Both the initial susceptibility and the saturation magnetization are lower than for the bare magnetic cores, but still significant. Drug adsorption and release tests were performed on two anticancer drugs, namely 5-fluorouracil and doxorubicin. Both are found to adsorb on the particles, but only the latter appears to be released at a reasonable rate, which is found to be very slow for 5-fluorouracil.

Temperature effects on energy production by salinity exchange.

In recent years, the capacitance of the interface between charged electrodes and ionic solutions (the electric double layer) has been investigated as a source of clean energy. Charge is placed on the electrodes either by means of ion-exchange membranes or of an external power source. In the latter method, net energy is produced by simple solution exchange in open circuit, due to the associated decrease in the capacitance of the electric double layer. In this work, we consider the change in capacitance associated with temperature variations: the former decreases when temperature is raised, and, hence, a cycle is possible in which some charge is put on the electrode at a certain potential and returned at a higher one. We demonstrate experimentally that it is thus viable to obtain energy from electric double layers if these are successively contacted with water at different temperatures. In addition, we show theoretically and experimentally that temperature and salinity variations can be conveniently combined to maximize the electrode potential increase. The resulting available energy is also estimated.

Electro-optics of confined systems.

Confinement in microenvironments occurs in many natural systems and technological applications. However, little is known about the behaviour of the immersed nanoparticles. In this work we show that their diffusion, electro-orientation and electric field induced polarization can be determined through electric birefringence experiments. We analyze aqueous dispersions of silver nanowires and clay particles confined inside microdroplets. We have observed that confinement reduces the amount of particles that can be oriented by the external electric field. However, the polarizability of the oriented particles is not affected by the presence of the oil/water boundary, and it is the same as in unbounded media, which agrees with the fact that the electric polarization and related phenomena are short-ranged.

Electro-optical Study of the Anomalous Rotational Diffusion in Polymer Solutions.

Brownian diffusion of spherical nanoparticles is usually exploited to ascertain the rheological properties of complex media. However, the behavior of the tracer particles is affected by a number of phenomena linked to the interplay between the dynamics of the particles and polymer coils. For this reason, the characteristic lengths of the dispersed entities, depletion phenomena, and the presence of sticking conditions have been observed to affect the translational diffusion of the probes. On the other hand, the retardation effect of the host fluid on the rotational diffusion of nonspherical particles is less understood. We explore the possibility of studying this phenomenon by analyzing the electro-orientation of the particles in different scenarios in which we vary the ratio between the particle and polymer characteristic size, and the geometry of the particles, including both elongated and oblate shapes. We find that the Stokes-Einstein relation only applies if the radius of gyration of the polymer is much shorter than the particle size and when some repulsive interaction between both is present.

Maghemite functionalization for antitumor drug vehiculization.

In this paper we describe the preparation and characterization of magnetic nanocomposites designed for applications in targeted drug delivery. Combining superparamagnetic behavior with proper surface functionalization in a single entity makes it possible to have altogether controlled location and drug loading, and release capabilities. The colloidal vehicles consist of maghemite (γ-Fe2O3) cores surrounded by a gold shell through an intermediate silica coating. The external Au layer confers the particles a high degree of biocompatibility and reactive sites for the transported drug binding. In addition, it permits to take advantage of the strong optical resonance, making it easy to visualize the particles or even control their payload release through temperature changes. The results of the analysis of relaxivity demonstrate that these nanostructures can be used as T2 contrast agents in magnetic resonance imaging (MRI), but the magnetic cores will be mainly useful in manipulating the particles using external magnetic fields. We describe how optical absorbance and electrokinetic data provide a followup of the progress of the nanostructure formation. Additionally, these techniques, together with confocal microscopy, are employed to demonstrate that the component nanoparticles are capable of loading significant amounts of the antitumor drug doxorubicin, very efficient in the chemotherapy of a wide range of tumors. Colon adenocarcinoma cells were used to test the in vitro release capabilities of the drug-loaded nanocomposites.

Hydrolysis versus ion correlation models in electrokinetic charge inversion: establishing application ranges.

In this article, we investigate experimentally a wide range of situations where charge inversion (i.e., overcompensation of the surface charge of a colloidal particle by the countercharge) can occur. To that end, the electrophoretic mobility of sodium montmorillonite, silica, and polystyrene latex as functions of pH and concentration of different salts is presented, and conditions are established where charge inversion occurs. The reason for this study is to provide experimental evidence for distinguishing between two existing models for the explanation of charge inversion. One of these is the specific adsorption of ions located in the Stern layer in combination with a Gouy-Chapman diffuse part of the double layer. The other ion-correlation theories explain the phenomenon in terms of purely physical arguments based on Coulombic pair interactions between ions and surface charges and on excluded volume effects. In distinguishing between these two interpretations, the influence of the pH plays a central role because of its effect on the hydrolysis of multivalent cations. In our experiments, it is found that although 1-2 and 2-2 electrolytes provoke a decrease in the absolute values of the electrophoretic mobilities when their concentration in solution is increased, they never lead to charge inversion, whatever the surface charge or the pH. However, in the case of salts of trivalent cations, electrokinetic charge reversal is often observed above a certain critical electrolyte concentration. In addition, the extent of overcharging increases when the concentration is raised above the critical value. This trend occurs for any system in which the surface charge is pH-independent, as in polystyrene latex and montmorillonite. Most of the results presented here are compatible with the specific adsorption of hydrolyzed metal ions as the main driving force for charge inversion. At low pH, when the hydrolysis of trivalent cations is likely to be absent, overcharging can be attributed to ion correlation effects.

Electrophoresis and dielectric dispersion of spherical polyelectrolyte brushes.

Spherical polyelectrolyte brushes (SPBs) consist of a rigid core on which polyelectrolyte chains are grafted in such a way that in certain conditions (low ionic strength and high charge of the chains) the polymer chains extend radially toward the liquid medium. Because of the hairy-like structure of the polymer brushes, the typical soft-particle approach used for explaining the behavior of polyelectrolyte-coated particles must be modified, using the assumptions that the density of charged segments in the polymer chains decreases with the squared distance to the rigid core surface and that the same happens to the friction between the brushes and the surrounding fluid. Interest in clarifying the electrokinetics of these systems is not just academic. It has recently been found experimentally (Jiménez et al., Soft Matter 2011, 7, 3758-3762) that the response of concentrated suspensions of spherical polyelectrolyte brushes in the presence of alternating electric fields shows a number of unexpected features. Both dielectric and dynamic electrophoretic mobility spectra (respectively, dependences of the electric permittivity and the AC electrophoretic mobility on the frequency of the applied field) showed very special aspects, with giant values of the mobility and an unusually strong dielectric relaxation in the kHz region. In the present paper we give a full account of the electrodynamics of such systems, based on a cell model for describing the hydrodynamic and electrical interactions between the particles. It is found that the low-frequency dynamic mobility of SPBs is much higher than that of rigid particles of comparable size and charge, making any interpretation based on zeta potential estimations of very limited applicability. The very characteristic feature of SPBs in concentrated suspensions, namely, the enhanced alpha relaxation, can be explained by considering an adequate description of the field-induced perturbations in the counterion and co-ion concentrations, well developed both outside and inside the soft layer in the case of brush-coated particles. It can be also pointed out that the dynamic electrophoretic mobility of SPBs increases with the volume fraction of particles, as a consequence of the large thickness of the brush. Predictions are also shown for the effects of friction coefficient and charge of the polyelectrolyte layer. The results compare well with experimental spectra of the dynamic mobility and electric permittivity of moderately concentrated suspensions of SPBs consisting of a 50 nm polystyrene core with grafted poly(styrene sulfonate) chains some 140 nm in length.

Electro-orientation of Ag nanowires in viscoelastic fluids.

In this work we study the electro-orientation (through electric birefringence experiments) of silver nanowires in polymer solutions eventually capable of forming gel networks. Information on the structure of the polymer solution is obtained by evaluating the electro-orientation of the nanowires. It is found that in presence of poly(ethylene oxide), Kerr's law (birefringence proportional to the square of the field) is fulfilled, and the randomization process after switching off the external field is purely diffusive, controlled by the viscosity of the Newtonian polymer solution. In the case of (gelating) sodium alginate solutions, measuring at larger distances from the bottom (where the source of cross-linking Ca2+ ions is deposited) means a smaller degree of cross-linking, and a less stiff gel. In fact, it is found that after a certain time the birefringence signal gets frozen at the bottom, indicating that a gel network is formed which hinders particle orientation. The viscosity deduced up to that point agrees well with rheological determinations, with increasing deviations found at longer times due to the inhomogeneous gel formation. This process has an interesting consequence on birefringence response: Kerr's law fails to be fulfilled, appearing a "yield" applied electric field, larger the longer the time after preparation.

Combined Magnetic Hyperthermia and Photothermia with Polyelectrolyte/Gold-Coated Magnetic Nanorods.

Magnetite nanorods (MNRs) are synthesized based on the use of hematite nanoparticles of the desired geometry and dimensions as templates. The nanorods are shown to be highly monodisperse, with a 5:1 axial ratio, and with a 275 nm long semiaxis. The MNRs are intended to be employed as magnetic hyperthermia and photothermia agents, and as drug vehicles. To achieve a better control of their photothermia response, the particles are coated with a layer of gold, after applying a branched polyethyleneimine (PEI, 2 kDa molecular weight) shell. Magnetic hyperthermia is performed by application of alternating magnetic fields with frequencies in the range 118-210 kHz and amplitudes up to 22 kA/m. Photothermia is carried out by subjecting the particles to a near-infrared (850 nm) laser, and three monochromatic lasers in the visible spectrum with wavelengths 480 nm, 505 nm, and 638 nm. Best results are obtained with the 505 nm laser, because of the proximity between this wavelength and that of the plasmon resonance. A so-called dual therapy is also tested, and the heating of the samples is found to be faster than with either method separately, so the strengths of the individual fields can be reduced. Due to toxicity concerns with PEI coatings, viability of human hepatoblastoma HepG2 cells was tested after contact with nanorod suspensions up to 500 µg/mL in concentration. It was found that the cell viability was indistinguishable from control systems, so the particles can be considered non-cytotoxic in vitro. Finally, the release of the antitumor drug doxorubicin is investigated for the first time in the presence of the two external fields, and of their combination, with a clear improvement in the rate of drug release in the latter case.

Electrophoretic characterization of insulin growth factor (IGF-1) functionalized magnetic nanoparticles.

The synthesis of composite nanoparticles consisting of a magnetite core coated with a layer of the hormone insulin growth factor 1 (IGF-1) is described. The adsorption of the hormone in the different formulations is first studied by electrophoretic mobility measurements as a function of pH, ionic strength, and time. Because of the permeable character expected for both citrate and IGF-1 coatings surrounding the magnetite cores, an appropriate analysis of their electrophoretic mobility must be addressed. Recent developments of electrokinetic theories for particles covered by soft surface layers have rendered possible the evaluation of the softness degree from raw electrophoretic mobility data. In the present contribution, the data are quantitatively analyzed based on the theoretical model of the electrokinetics of soft particles. As a result, information is obtained on both the thickness and the charge density of the surrounding layer. It is shown that IGF-1 adsorbs onto the surface of citrate-coated magnetite nanoparticles, and adsorption is confirmed by dot-blot analysis. In addition, it is also demonstrated that the external layer of IGF-1 exerts a shielding effect on the surface charge of citrate-magnetite particles, as suggested by the mobility reduction upon contacting the particles with the hormone. Aging effects are demonstrated, providing an electrokinetic fingerprint of changes in adsorbed protein configuration with time.

Coagulation changes to systemic acidosis and bicarbonate correction in swine.

BACKGROUND: As part of our overall interest in the mechanisms and treatment related to the development of the lethal triad of hypothermia, acidosis, and coagulopathy seen in trauma patients, the purpose of this study was to determine whether acidosis, inducible either by HCl infusion or hemorrhage/hypoventilation, leads to coagulopathy, and if correction of the acidosis will alleviate this coagulopathy. METHODS: In two separate experiments, acidosis was induced in anesthetized swine by (1) HCl infusion (n = 10) or (2) hemorrhage/hypoventilation (n = 8). Arterial blood samples were taken before HCl infusion or hemorrhage (arterial pH 7.4), after HCl infusion or hemorrhage (pH 7.1), and after bicarbonate infusion to return pH to 7.4. Arterial pH and blood gases were measured every 15 minutes. RESULTS: Acidosis (arterial pH 7.1) led to a hypocoagulation as measured by several coagulation parameters. In both experiments, acidosis was associated with a significant decrease in the maximum strength of the clot and the rate at which the clot formed. There was a significant decrease in endogenous thrombin potential and maximum thrombin concentration after acidosis in both groups (thrombin generation assay). However, the activated clotting time, prothrombin time, and activated partial thromboplastin time were significantly elevated only in the HCl-infused group. Fibrinogen concentration and platelet count were significantly reduced in both groups after acidosis. The hypocoagulation that was induced by either hemorrhage/hypoventilation or HCl infusion was not immediately corrected after returning pH to 7.4 with bicarbonate injection. CONCLUSIONS: These data suggest that acidosis induced by HCl infusion or by hemorrhage/hypoventilation leads to hypocoagulation. Simple correction of the arterial pH with bicarbonate is not sufficient to correct this coagulopathy.

Effect of hemodilution on coagulation and recombinant factor VIIa efficacy in human blood in vitro.

BACKGROUND: This study evaluates the effect of hemodilution by various common resuscitation fluids, and the efficacy of activated recombinant factor VII (rFVIIa) on coagulation parameters in human blood in vitro. METHODS: Samples from normal healthy volunteers (n = 9) were hemodiluted from 0% to 90% with normal saline, or 0%, 40%, 60%, and 80% with 5% albumin, Hespan, Hextend, normal saline, or lactated Ringer's, and incubated at 37°C ± 1°C for 30 minutes with and without rFVIIa (1.26 µg/mL). RESULTS: There was a strong correlation between the dilution of hemoglobin (Hb), platelets, or fibrinogen and coagulation parameters. Hemodilution 0% to 90% changed coagulation parameters (prothrombin time [PT], activated partial thromboplastin time [aPTT], and thromboelastography) in an exponential fashion; the greatest changes occurred after hemodilution lowered Hb <6 mg/dL, platelet count < 100,000/mm(3), and fibrinogen concentration <200 mg/dL. PT and aPTT were significantly prolonged after 60% and 80% dilution for all fluids. Hemodilution of 60% and 80% significantly decreased clot strength (maximum amplitude) and the kinetics of clot development (α angle) and increased the clot formation time (K). Hemodilution with Hextend and Hespan decreased maximum amplitude and α angle >5% albumin, lactated Ringer's, or normal saline. rFVIIa significantly improved PT at 60% and 80% dilutions, and aPTT at 80% dilution. There was a significant effect of dilution, but not fluid type, on the efficacy of rFVIIa to change PT and aPTT, and the onset of clotting (R). CONCLUSIONS: We have strong in vitro evidence that Hb <6 mg/dL, platelet count <100,000/mm(3), and fibrinogen concentration <200 mg/dL can be used as indexes of hemodilution-induced coagulopathy. This study also shows that Hextend and Hespan tend to decrease the clotting ability >5% albumin or the crystalloids. rFVIIa significantly decreased PT at all dilutions and aPTT at the highest dilution. The effectiveness of rFVIIa on PT and aPTT was significantly affected by the degree of dilution, but not by the type of fluid.