Indomethacin functionalised poly(glycerol adipate) nanospheres as promising candidates for modified drug release.

The linear polyester poly(glycerol adipate) (PGA) with its free pendant hydroxyl groups was covalently grafted with indomethacin which yields polymeric prodrugs. It was possible to produce nanospheres with narrow particle size distribution of these polymer-drug conjugates with an optimized interfacial deposition method. Nanospheres were characterized by zeta potential measurements, dynamic light scattering, electron microscopy and nanoparticle tracking analysis. Moreover, cell viability studies and cytotoxicity tests in three different cell lines were carried out showing low toxicity for three different degrees of grafting. In addition, the nanospheres had (in contrast to the free drug) low hemolytic activity in vitro. Release studies of nanodispersions are challenging. The use of a specially developed setup with highly porous aluminum oxide membranes enabled us to overcome problems associated with other setups (e.g. dialysis membranes). A slow and controlled release profile without any burst was observed over 15 days. The results indicate that indomethacin-PGA conjugates can be formulated successfully as nanospheres with the desired characteristics of small size with narrow distribution, controlled drug release and low toxicity. The newly developed particles have the potential to improve the therapy of inflammation and associated diseases.

Poly(glycerol adipate) - indomethacin drug conjugates - synthesis and in vitro characterization.

The linear biodegradable polyester poly(glycerol adipate) (PGA) was synthesized via enzymatic polycondensation using lipase B from Candida antarctica (CAL-B). Every monomer unit of PGA possesses a pendant hydroxyl group which is responsible for the hydrophilic character and moisture swelling. These OH groups were esterified to different degrees with the anti-inflammatory drug indomethacin in order to create a prodrug with a pH-sensitive linker for modified drug release. The structure of the conjugates was determined via ATR FT-IR spectroscopy, NMR spectroscopy, GPC and UV/VIS spectroscopy. The physical properties of polymers with different drug load were investigated using DSC, contact angle measurements and oscillatory rheology. Drug release was monitored over one month in vitro. A very slow, but continuous release was observed in PBS. Slightly acidic conditions and lipase activity are accelerating the indomethacin release. Therefore, poly(glycerol adipate) - indomethacin conjugates are promising prodrugs for the local sustained release of indomethacin.

Effects of circuit resistance training and timely protein supplementation on exercise-induced fat oxidation in tetraplegic adults.

BACKGROUND: Substrate utilization during exercise in persons with spinal cord injury (SCI) remains poorly defined. PURPOSE: To investigate effects of circuit resistance training (CRT) and timing of protein supplementation (PS) on fuel utilization in persons with tetraplegia. METHODS: Eleven individuals with chronic tetraplegia underwent 6 months of CRT 3 times weekly. Five randomly assigned participants received immediate PS (iPS) administered in split doses prior to and following all exercise sessions. Other participants consumed a matched dose of PS that was delayed until 24 hours post-exercise (dPS). Participants underwent a maximal graded exercise test (GXT) to volitional exhaustion at 4 conditioning time points: 3 months before (-3mo), at the beginning of (0mo), 3 months into (3mo), and 6 months following (6mo) the CRT conditioning program. Respiratory measures were continuously obtained throughout the GXT via open-circuit spirometry. Fuel utilization and energy expenditure were computed from the respiratory data. RESULTS: The differences in changes in substrate utilization between the PS groups were not significant as determined by the interaction of PS group and conditioning time point, F (3, 27) = 2.32, P = .098, η(2) P = .205. Maximal absolute fat oxidation did not change significantly from 0 to 6mo (mean difference, 0.014 ± 0.031 g/min; P = .170), and fat oxidation remained low never exceeding an average of 0.10 ± 0.09 g/min for any given exercise intensity. CONCLUSION: Maximum fat utilization during exercise and fat utilization at matched exercise intensities were not increased in persons with tetraplegia, independent of PS, and levels of fat oxidation remained low after training.

Aerobic training does not alter CRP in apparently healthy, untrained men.

AIM: Regular aerobic exercise may reduce cardiovascular disease (CVD) risk by lowering the concentration of inflammatory markers, such as C-reactive protein (CRP). While studies in diseased populations have shown significant decreases in CRP concentrations with regular aerobic training, little has been conclusively determined regarding the effects of aerobic training on CRP concentrations in apparently healthy, untrained populations. Aim of the study was to examine the effects of a 17-wk half marathon training program (TP) on CRP concentrations, aerobic fitness, and body composition in apparently healthy, untrained men. METHODS: Twenty men (29.3±1.0 y) enrolled as training subjects (TRN) in a 17-wk half marathon TP. An additional 22 men (27.8±1.4 y) served as controls (CON). Fasting blood samples were taken at four time points over the TP and were analyzed for CRP and interleukin-6 (IL-6) concentrations. Aerobic capacity (VO2max) and body fat percent (BF%) were measured before and after the TP. RESULTS: No significant post-training changes in CRP (P=0.70) or IL-6 concentrations (P=0.67) were seen in TRN as a result of the TP, despite significant improvements in VO2max (42.2±1.9 ml∙kg-1∙min⁻¹, P<0.0001) and significant reductions in resting heart rate (P=0.004), BF% (P=0.03), and body mass index (BMI, P=0.05). No significant changes in CRP, VO2max, BMI, or BF% were detected in CON over time. CONCLUSION: Regular aerobic training does not appear to affect CRP concentrations in apparently healthy, untrained men despite significant improvements in bodyweight, BF%, BMI, and VO2max.

Alginate encapsulation of genetically engineered mammalian cells: comparison of production devices, methods and microcapsule characteristics.

PRIMARY OBJECTIVE: Microencapsulation is a novel method for in vivo immunoprotection of genetically engineered mammalian cells. This study aimed at optimizing alginate/poly-l-lysine/alginate (APA) microencapsulation of mammalian cells in small size (< 300 microm) hollow core microcapsules and at evaluating some of the physical characteristics of APA microcapsules produced by different devices and with different alginate preparations. METHODS AND PROCEDURES: Alginate preparations with higher or lower viscosity were used with three different methods: (i) vibrating nozzle, (ii) coaxial gas flow extrusion (AirJet) and (iii) laminar jet break-up (JetCutter). Parameters and device settings for the production of microcapsules with specific characteristics were defined for all three methods. Mechanical stability of APA microcapsules and cell viability of encapsulated cells were investigated in long-term culture and in an animal model. MAIN RESULTS: Uniform spherical beads with a mean diameter < 300 microm could be produced by all three encapsulation methods. For the production of uniform microcapsules with a small diameter (< 300 microm) the vibrating nozzle technique required a relatively low viscosity of alginate (< 0.2 Pa/s), while the AirJet and JetCutter devices performed equally well with alginate of higher viscosity. In all cases, alginate with a lower molar mass was inferior to higher molar mass alginate in terms of mechanical stability of the microcapsules. Microcapsules with optimized mechanical properties were injected intravascularly in rats and shown to maintain their shape and the viability of encapsulated cells. CONCLUSIONS: The described methods and devices are able to produce APA microcapsules of small size and uniform shape which are mechanically stable in culture and may maintain the viability of the enclosed cells over extended periods of time. These microcapsules seem to be suitable for further therapeutic studies in an animal model of human disease.

High throughput encapsulation of murine fibroblasts in alginate using the JetCutter technology.

The JetCutter technology originally developed for high-throughput encapsulation of particles and substances into small beads was applied in this study for the entrapment of mammalian cells in alginate beads. In contrast to other established techniques such as the air jet droplet generation or laminar jet break-up, the JetCutter is capable of working with highly viscous fluids necessary for the production of stable beads based on hydrogels. A 1.5% (w/v) sodium alginate solution containing 2.0 x 106 murine fibroblasts/ml was processed under good manufacturing practice (GMP) conditions to beads with a mean diameter of 320 microm. The production capacity of the JetCutter technology was 5200 beads/s or to approximately 330 ml bead suspension per h. Beads were coated with poly-L-lysine and with an additional alginate layer to produce hollow microcapsules containing living cells. The influence of this method of encapsulation on the cell viability and morphology was investigated by light microscopic techniques. Encapsulated cells showed unchanged rates of proliferation and preserved morphology. They were able to survive in culture for extended periods of time. In conclusion, the JetCutter technology seems to be well suitable for alginate bead encapsulation of living mammalian cells.

Synthesis of microbial poly(beta-hydroxybutyrate) modified with oligo(pentaerythritol ethoxylate) by Ralstonia eutropha.

Poly(beta-hydroxybutyrate) (PHB) modified with different amounts of pentaerythritol ethoxylate (PEE) has been synthesized using Ralstonia eutropha. The growth kinetics and the synthesis of PHB in the presence of PEE were modeled using appropriate differential equations for the mass balance of the two-stage process. The influence of PEE addition on the morphology of PHB was studied by various microscopic and scattering techniques. Light microscopic and wide-angle X-ray measurements indicated that the addition of PEE had a nucleating effect on the crystallization of PHB. The spherulite growth rate was widely independent of the PEE addition. The lamellae of PHB became more disordered when PEE was added as demonstrated by atomic force microscopy. Furthermore, small-angle X-ray data indicated a decrease in the long period with increasing PEE content of the modified PHB.