Production of a novel medium chain length Poly(3-hydroxyalkanoate) using unprocessed biodiesel waste and its evaluation as a tissue engineering scaffold

This study demonstrated the utilisation of unprocessed biodiesel waste as a carbon feedstock for Pseudomonas mendocina CH50, for the production of PHAs. A PHA yield of 39.5% CDM was obtained using 5% (v/v) biodiesel waste substrate. Chemical analysis confirmed that the polymer produced was poly(3-hydroxyhexanoate-co-3-hydroxyoctanoate-co-3- hydroxydecanoate-co-3-hydroxydodecanoate) or P(3HHx-3HO-3HD-3HDD). P(3HHx-3HO- 3HD-3HDD) was further characterised and evaluated for its use as a tissue engineering scaffold (TES). This study demonstrated that P(3HHx-3HO-3HD-3HDD) was biocompatible with the C2C12 (myoblast) cell line. In fact, the % cell proliferation of C2C12 on the P(3HHx-3HO-3HD-3HDD) scaffold was 72% higher than the standard tissue culture plastic confirming that this novel PHA was indeed a promising new material for soft tissue engineering

Binary Polyhydroxyalkanoate Systems for Soft Tissue Engineering

Progress in tissue engineering is dependent on the availability of suitable biomaterials. In an effort to overcome the brittleness of poly(3-hydroxybutyrate), P(3HB), a natural biodegradable polyester, and widen its biomedical applications, plasticising of P(3HB) with oligomeric substances of related structure has been studied. A biosynthesised medium-chain-length polyhydroxyalkanoate (mcl-PHA) copolymer, the plasticizer precursor, was obtained using vegetable waste frying oil as a sole carbon source. The mcl-PHA was transformed into an oligomeric derivative by acid hydrolysis. The plasticising effect of the oligomeric mcl-PHA on P(3HB) was studied via characterisation of thermal and mechanical properties of the blends in the course of ageing at ambient conditions. Addition of oligomeric mcl-PHA to P(3HB) resulted in softer and more flexible materials based entirely on PHAs. It was shown that the oligomeric mcl-PHA transformed highly crystalline P(3HB) into materials with a dominant amorphous phase when the content of oligomeric mcl-PHA exceeded 10wt%. In vitro biocompatibility studies of the new binary PHA materials showed high viability and proliferation of C2C12 myoblast cells. Thus, the proposed approach for P(3HB) plasticisation has the potential for the generation of more pliable biomaterials based on P(3HB) which can find application in unique soft tissue engineering applications where a balance between stiffness, tensile strength and ductility is required

Effects of therapeutic ultrasound on osteoblast gene expression

Ultrasound (US) is commonly used as a physiotherapy aid for a number of types of injury to soft connective tissues and for fracture healing. However, the precise effects of therapeutic US on tissue healing processes are not clearly understood, although they are likely to involve changes in key cellular functions. The present study has therefore examined the effects of several US intensity levels on the activity of two bone-associated proteins, alkaline phosphatase (ALP) and osteopontin (OP) in a human cell line, MG63, using RT-PCR. ALP showed progressively higher expression with increasing US intensities, whereas OP responded differently, showing down-regulation at 120 mW/cm2, the lowest US exposure. OP expression was considerably less affected overall compared with the relative response of ALP to the same US doses. The results show that there is a differential response to therapeutic levels of US, since ALP and OP clearly exhibited gene-specific response profiles. These findings suggest that modifying the parameters of US exposure could be used to improve repair and regeneration processes and enhance the clinical efficacy of implanted biomaterials for tissue engineering

Unidirectional Neuronal Cell growth and Differentiation on Aligned Polyhydroxyalkanoate Blend Microfibres with Varying Diameters

Polyhydroxyalkanoates (PHAs) are a family of prokaryotic-derived biodegradable and biocompatible natural polymers known to exhibit neuroregenerative properties. In this work, poly(3-hydroxybutyrate), P(3HB) and poly(3-hydroxyoctanoate), P(3HO), have been combined to form blend fibres for directional guidance of neuronal cell growth and differentiation. A 25:75 P(3HO)/P(3HB) blend (PHA blend) was used for the manufacturing of electrospun fibres as resorbable scaffolds to be used as internal guidance lumen structures in nerve conduits. The biocompatibility of these fibres was studied using neuronal and Schwann cells. Highly aligned and uniform fibres with varying diameters were fabricated by controlling electrospinning parameters. The resulting fibre diameters were 2.4 ± 0.3 µm, 3.7 ± 0.3 µm and 13.5 ± 2.3 µm for small, medium and large diameter fibres respectively. The cell response to these electrospun fibres was investigated with respect to growth and differentiation. Cell migration observed on the electrospun fibres showed topographical guidance in accordance with the direction of the fibres. The correlation between fibre diameter and neuronal growth under two conditions; individually and in co-culture with Schwann cells was evaluated. Results obtained from both assays revealed that all PHA blend fibre groups were able to support growth and guide aligned distribution of neuronal cells and there was a direct correlation between the fibre diameter and neuronal growth and differentiation. This work has led to the development of a family of unique biodegradable and highly biocompatible 3D substrates capable of guiding and facilitating the growth, proliferation and differentiation of neuronal cells as internal structures within nerve conduits

Nerve tissue engineering using blends of poly(3-hydroxyalkanoates) for peripheral nerve regeneration

The only types of polyhydroxyalkanoates (PHAs) that have been explored for use in nerve regeneration are poly(3‐hydroxybutyrate), P(3HB), and poly(3‐hydroxybutyrate‐co‐3‐hydroxyhexanoate) (P(3HB‐co‐3HHx)). However, nerve regeneration induced by these PHAs is inferior to that of autologous nerve grafting. The aim of this work was to study novel PHA blends as resorbable biomaterials for the manufacture of nerve guidance conduits. PHA blend films with varying ratios of poly(3‐hydroxyoctanoate)/poly(3‐hydroxybutyrate) (P(3HO)/P(3HB)) were produced using the solvent‐casting method. Neat films of P(3HO) and P(3HB), along with 25:75, 50:50, and 75:25 blend films of P(3HO)/P(3HB), were characterized with respect to chemical, material, and biological properties. On surface analysis, the blends exhibited higher values of roughness compared with the neat films. The differential scanning calorimetry characterization of the blends confirmed that P(3HO) and P(3HB) formed immiscible blends. FTIR and XRD analysis of the blends showed a decrease in crystallinity along with an increase of the proportion of P(3HO) . However, an increase in the stiffness of the blends was observed when the proportion of P(3HB) increased. Although all of the blends were biocompatible with NG108‐15 neuronal cells, the 25:75 P(3HO)/P(3HB) blend showed significantly better support for growth and differentiation of these cells. The mechanical properties of PHA blends correspond to the reported properties of peripheral nerves. Therefore, they could serve as base material for the manufacture of nerve guidance conduits

Modulation of neuronal cell affinity of composites scaffolds based on polyhydroxyalkanoates and bioactive glasses

Biocompatibility and neuron regenerating properties of various bioactive glass (BG)/Polyhydroxyalkanoate (PHA) blend composites were assessed in order to study their suitability for peripheral nerve tissue applications, specifically as lumen structures for nerve guidance conduits (NGCs). BG/PHA blend composites were fabricated using Bioactive glass® 45S5 (BG1) and BG 1393 (BG2) with the 25:35 poly(3-hydroxyoctanoate/poly3-hydroxybutyrate), 25:75 P(3HO)/P(3HB) blend (PHA blend). Various concentrations of each BG (0.5, 1.0 and 2.5 wt%) were used to determine the effect of BG on neuronal growth and differentiation, in single culture using NG108-15 neuronal cells and in a co-culture along with RN22 Schwann cells. NG108-15 cells exhibited good growth and differentiation on all the PHA blend composites showing that both BGs have good biocompatibility at 0.5, 1.0 and 2.5 wt% within the PHA blend. The Young's modulus values displayed by all the PHA blend/BG composites ranged from 385.6 MPa to 1792.6 MPa, which are able to provide the required support and protective effect for regeneration of peripheral nerves. More specifically, the tensile strength obtained in the PHA blend/BG1 (1.0 wt%) (10.0 ± 0.6 MPa) was found to be similar to that of rabbit peroneal nerve. This composite also exhibited the best biological performance in supporting growth and neuronal differentiation among all the substrates. The neurite extension on this composite was found to be remarkable with the neurites forming a complex connection network

Characterisation of phosphate coacervates for potential biomedical applications

In this study, amorphous (Na2O)x(CaO)0.50−x(P2O5)0.50·yH2O (where x = ∼0.15 and y = ∼3) samples were prepared by a coacervate method. Thermal analysis showed that two types of water molecules were present in the coacervate structures: one type loosely bound and the other part of the phosphate structure. Structural studies using Fourier transform infrared spectroscopy (FTIR) and X-ray total diffraction revealed the samples to have very similar structures to melt-quenched glasses of comparable composition. Furthermore, no significant structural differences were observed between samples prepared using calcium nitrate as the calcium source or those prepared from calcium chloride. A sample containing ∼1 mol% Ag2O was prepared to test the hypothesis that calcium phosphate coacervate materials could be used as delivery agents for antibacterial ions. This sample exhibited significant antibacterial activity against the bacterium Psuedomonas aeruginosa. FTIR data revealed the silver-doped sample to be structurally akin to the analogous silver-free sample

Distribution modelling and statistical phylogeography: an integrative framework for generating and testing alternative biogeographical hypotheses

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/73644/1/j.1365-2699.2007.01814.x.pd

Theileria parasites subvert E2F signaling to stimulate leukocyte proliferation

Intracellular pathogens have evolved intricate mechanisms to subvert host cell signaling pathways and ensure their own propagation. A lineage of the protozoan parasite genus Theileria infects bovine leukocytes and induces their uncontrolled proliferation causing a leukemia-like disease. Given the importance of E2F transcription factors in mammalian cell cycle regulation, we investigated the role of E2F signaling in Theileria-induced host cell proliferation. Using comparative genomics and surface plasmon resonance, we identified parasite-derived peptides that have the sequence-specific ability to increase E2F signaling by binding E2F negative regulator Retinoblastoma-1 (RB). Using these peptides as a tool to probe host E2F signaling, we show that the disruption of RB complexes ex vivo leads to activation of E2F-driven transcription and increased leukocyte proliferation in an infection-dependent manner. This result is consistent with existing models and, together, they support a critical role of E2F signaling for Theileria-induced host cell proliferation, and its potential direct manipulation by one or more parasite proteins

PYTHIA 6.4 Physics and Manual

The PYTHIA program can be used to generate high-energy-physics `events', i.e. sets of outgoing particles produced in the interactions between two incoming particles. The objective is to provide as accurate as possible a representation of event properties in a wide range of reactions, within and beyond the Standard Model, with emphasis on those where strong interactions play a role, directly or indirectly, and therefore multihadronic final states are produced. The physics is then not understood well enough to give an exact description; instead the program has to be based on a combination of analytical results and various QCD-based models. This physics input is summarized here, for areas such as hard subprocesses, initial- and final-state parton showers, underlying events and beam remnants, fragmentation and decays, and much more. Furthermore, extensive information is provided on all program elements: subroutines and functions, switches and parameters, and particle and process data. This should allow the user to tailor the generation task to the topics of interest.Comment: 576 pages, no figures, uses JHEP3.cls. The code and further information may be found on the PYTHIA web page: http://www.thep.lu.se/~torbjorn/Pythia.html Changes in version 2: Mistakenly deleted section heading for "Physics Processes" reinserted, affecting section numbering. Minor updates to take into account referee comments and new colour reconnection option