SN-38 active loading in poly(lactic-co-glycolic acid) nanoparticles and assessment of their anticancer properties on COLO-205 human colon adenocarcinoma cells.

SN-38 is a highly effective drug against many cancers. The development of an optimal delivery system for SN-38 is extremely challenging due to its low solubility and labile lactone ring. Herein, SN-38 encapsulated in poly(D,L-lactide-co-glycolide) nanoparticles (NPs) is introduced to enhance its solubility, stability and cellular uptake. SN-38-loaded NPs prepared by spontaneous emulsification solvent diffusion (SESD) method had an average diameter of 310 nm, a zeta potential of -9.69 mV and a loading efficiency of 71%. They were able to protect the active lactone ring of SN-38 against inactivation under physiological condition. A colorectal adenocarcinoma cell line (COLO-205) was used to assess the NPs effects on cytotoxicity and cellular uptake. Result showed a significant decreased cell proliferation and cell apoptosis. These results suggest that these SN-38-loaded NPs can be an effective delivery system for the treatment of colon cancer and potentially for other types of cancers.

Magnetic resonance imaging-based radiation-absorbed dose estimation of 166Ho microspheres in liver radioembolization.

PURPOSE: To investigate the potential of magnetic resonance imaging (MRI) for accurate assessment of the three-dimensional (166)Ho activity distribution to estimate radiation-absorbed dose distributions in (166)Ho-loaded poly (L-lactic acid) microsphere ((166)Ho-PLLA-MS) liver radioembolization. METHODS AND MATERIALS: MRI, computed tomography (CT), and single photon emission CT (SPECT) experiments were conducted on an anthropomorphic gel phantom with tumor-simulating gel samples and on an excised human tumor-bearing liver, both containing known amounts of (166)Ho-PLLA-MS. Three-dimensional radiation-absorbed dose distributions were estimated at the voxel level by convolving the (166)Ho activity distribution, derived from quantitative MRI data, with a (166)Ho dose point-kernel generated by MCNP (Monte Carlo N-Particle transport code) and from Medical Internal Radiation Dose Pamphlet 17. MRI-based radiation-absorbed dose distributions were qualitatively compared with CT and autoradiography images and quantitatively compared with SPECT-based dose distributions. Both MRI- and SPECT-based activity estimations were validated against dose calibrator measurements. RESULTS: Evaluation on an anthropomorphic phantom showed that MRI enables accurate assessment of local (166)Ho-PLLA-MS mass and activity distributions, as supported by a regression coefficient of 1.05 and a correlation coefficient of 0.99, relating local MRI-based mass and activity calculations to reference values obtained with a dose calibrator. Estimated MRI-based radiation-absorbed dose distributions of (166)Ho-PLLA-MS in an ex vivo human liver visually showed high correspondence to SPECT-based radiation-absorbed dose distributions. Quantitative analysis revealed that the differences in local and total amounts of (166)Ho-PLLA-MS estimated by MRI, SPECT, and the dose calibrator were within 10%. Excellent agreement was observed between MRI- and SPECT-based dose-volume histograms. CONCLUSIONS: Quantitative MRI was demonstrated to provide accurate three-dimensional (166)Ho-PLLA-MS activity distributions, enabling localized intrahepatic radiation-absorbed dose estimation by convolution with a (166)Ho dose point-kernel for liver radioembolization treatment optimization and evaluation.

Assessment of material by-product fate from bioresorbable vascular scaffolds.

Fully bioresorbable vascular scaffolds (BVS) are attractive platforms for the treatment of ischemic artery disease owing to their intrinsic ability to uncage the treated vessel after the initial scaffolding phase, thereby allowing for the physiological conditioning that is essential to cellular function and vessel healing. Although scaffold erosion confers distinct advantages over permanent endovascular devices, high transient by-product concentrations within the arterial wall could induce inflammatory and immune responses. To better understand these risks, we developed in this study an integrated computational model that characterizes the bulk degradation and by-product fate for a representative BVS composed of poly(L-lactide) (PLLA). Parametric studies were conducted to evaluate the relative impact of PLLA degradation rate, arterial remodeling, and metabolic activity on the local lactic acid (LA) concentration within arterial tissue. The model predicts that both tissue remodeling and PLLA degradation kinetics jointly modulate LA fate and suggests that a synchrony of these processes could minimize transient concentrations within local tissue. Furthermore, simulations indicate that LA metabolism is a relatively poor tissue clearance mechanism compared to convective and diffusive transport processes. Mechanistic understanding of factors governing by-product fate may provide further insights on clinical outcome and facilitate development of future generation scaffolds.

Comparative assessment of in vitro release kinetics of calcitonin polypeptide from biodegradable microspheres.

The objective of our study was to compare the in vitro release kinetics of a sustained-release injectable microsphere formulation of the polypeptide drug, calcitonin (CT), to optimize the characteristics of drug release from poly-(lactide-co-glycolide) (PLGA) copolymer biodegradable microspheres. A modified solvent evaporation and double emulsion technique was used to prepare the microspheres. Release kinetic studies were carried out in silanized tubes and dialysis bags, whereby microspheres were suspended and incubated in phosphate buffered saline, sampled at fixed intervals, and analyzed for drug content using a modified Lowry protein assay procedure. An initial burst was observed whereby about 50% of the total dose of the drug was released from the microspheres within 24 hr and 75% within 3 days. This was followed by a period of slow release over a period of 3 weeks in which another 10-15% of drug was released. Drug release from the dialysis bags was more gradual, and 50% CT was released only after 4 days and 75% after 12 days of release. Scanning electron micrographs revealed spherical particles with channel-like structures and a porous surface after being suspended in an aqueous solution for 5 days. Differential scanning calorimetric studies revealed that CT was present as a mix of amorphous and crystalline forms within the microspheres. Overall, these studies demonstrated that sustained release of CT from PLGA microspheres over a 3-week period is feasible and that release of drug from dialysis bags was more predictable than from tubes.

Value of lactic acidosis in the assessment of the severity of acute cyanide poisoning.

OBJECTIVE: To test the hypothesis that plasma lactate concentrations could be of confirmatory value in patients with histories consistent with acute pure cyanide poisoning because immediate laboratory confirmation of suspected cyanide poisoning is rarely possible and because clinicians must rapidly decide whether to administer specific antidotes, which may have severe side effects. DESIGN: Retrospective clinical study. SETTING: An intensive care unit in a university-affiliated teaching hospital. PATIENTS: All acute cyanide-poisoned patients admitted to our intensive care unit, excluding fire victims, from 1988 to 1999. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Eleven patients were studied. Before antidotal treatment, the median plasma lactate concentration was 168 mg/dL, the median blood cyanide concentration was 4.2 mg/L. Using Spearman's test, there was a significant correlation between plasma lactate and blood cyanide concentrations ( =.74, =.017). Before antidotal treatment, plasma lactate concentration correlated positively with anion gap and inversely with systolic blood pressure, spontaneous respiratory rate, and arterial pH. During the course of cyanide poisonings, a plasma lactate concentration of >or=72 mg/d/L (8 mmol/L) was sensitive (94%) and moderately specific (70%) for a toxic blood cyanide concentration (>or=1.0 mg/L). The specificity was substantially improved in patients not receiving catecholamines (85%). CONCLUSIONS: The immediate and serial measurement of plasma lactate concentrations is useful in assessing the severity of cyanide poisoning.

Assessment of protein release kinetics, stability and protein polymer interaction of lysozyme encapsulated poly(D,L-lactide-co-glycolide) microspheres.

Using lysozyme as a model protein, this study investigated protein stability, protein--polymer interaction in different release media and their influence on protein release profile and in vitro--in vivo correlation. Lysozyme was microencapsulated into PLGA 50:50 by a double emulsion--solvent extraction/evaporation method. Protein stability, protein--PLGA adsorption and protein in vitro release were studied in various test media. Differential scanning calorimetry analysis showed lysozyme to be most conformationally stable in pH 4.0 acetate buffer with highest T(m) at 77.2 degree C and DeltaH(cal) 83.1 kcal/mol. Lysozyme exhibited good stability in pH 2.5 glycine buffer with T(m) at 63.8 degree C and DeltaH(cal) 69.9 kcal/mol. In pH 7.4 phosphate-buffered saline (PBS), lysozyme showed a trend toward aggregation when the temperature was elevated. When PLGA polymer was incubated with lysozyme in the various buffers, adsorption was found to occur in PBS only. The adsorption severely limited the amount of lysozyme available for release from microspheres, resulting in slow and incomplete release in PBS. In contrast, the release of the microspheres in acetate and glycine buffers was complete within 40 and 70 days, respectively. Radiolabeled lysozyme blood levels in rats from the microspheres correlated qualitatively well with in vitro release in glycine buffer as a release medium. This study suggests that protein stability and adsorption are critical factors controlling protein release kinetics and in vitro--in vivo correlation of PLGA microspheres.

Blood pH and lactate kinetics in the assessment of running endurance.

To study how two types of testing protocols for the determination of the [LA] and blood pH kinetics during running match each other, a group of nine runners participated in two testing protocols. The first protocol consisted of a sequences of 8 x 2000 m runs at constant speed, which was increased on different testing days. The first protocol was used for the determination of running speed (v), [LA], pH and heart rate (HR) corresponding to the Lactate Threshold (LT) and the Threshold of Acidosis (TA), both occurring at similar running speeds (4.21 +/- 0.44 and 4.22 +/- 0.40 m/s) (mean +/- SD) and HR (159 +/- 7 and 160 +/- 9 1/min), [LA] = 2.0 +/- 0.6 mmol/l and pH = 7.411 +/- 0.018. Maximal steady values for [LA] (maxLAss) and minimal steady values for pH (minpHss) obtained by the second protocol were higher (p < 0.01) according to running speed (4.62 +/- 0.38 and 4.66 +/- 0.38 m/s), HR (172 +/- 7 and 174 +/- 8 1/min) and [LA] (5.7 +/- 1.3 mmol/l) and lower according to pH (7.364 +/- 0.021), respectively. Unlike similar running speeds determined by LT and TA, the minimal steady pH level occurred at a slightly higher speed than the speed at maxLAss. Additionally, we found a drift of pH towards a resting level, when [LA] fluctuated around a steady level. Furthermore, the parameters of pH kinetics correlated better with the running speed of the 4000 m run that was taken as the parameter of short endurance performance, than those of [LA] kinetics. We conclude that these differences in [LA] and pH kinetics could serve to predict the capabilities of runners with respect to the two endurance performance types: long and short.