First-Order Theory of Rewriting for Linear Variable-Separated Rewrite Systems: Automation, Formalization, Certification.

The first-order theory of rewriting is decidable for linear variable-separated rewrite systems. We present a new decision procedure which is the basis of FORT, a decision and synthesis tool for properties expressible in the theory. The decision procedure is based on tree automata techniques and verified in Isabelle. Several extensions make the theory more expressive and FORT more versatile. We present a certificate language that enables the output of FORT to be certified by the certifier FORTify generated from the formalization, and we provide extensive experiments.

The influence of covalently linked and free polyethylene glycol on the structural and release properties of rhBMP-2 loaded microspheres.

The current clinical success of therapies with recombinant human Bone Morphogenetic Protein 2 (rhBMP-2) is limited due to inefficient delivery. The high doses applied have frequently been related to severe adverse effects such as tissue swelling, seroma, inflammatory effects and heterotopic ossification. The controlled delivery of lower doses is supposed to reduce adverse effect incidence as well as costs. In this study, novel polyethylene glycol-poly(lactic-co-glycolic acid) (PEG-PLGA) diblock copolymers were used to produce low dose controlled delivery vehicles for rhBMP-2. A method to fabricate a variety of microsphere formulations with a high encapsulation efficiency in high yields was developed. The influence of PEG as an inner phase cosolvent and linked PLGA as copolymer was investigated. Six different microsphere systems with varying PEG amounts in both core and shell were characterised thoroughly with respect to the specific properties of rhBMP-2. The particle size of the microspheres was investigated with both laser diffraction and environmental scanning electron microscopy. Higher PEG/PLGA ratios showed a tendency to increase in size and a wider distribution. Due to the low rhBMP-2 doses, a profound characterisation was very challenging. The growth factor was covalently attached to rhodamine B for the first time. Studies on drug distribution in the microspheres were performed by means of confocal laser scanning microscopy. The addition of PEG to the inner phase was found to impair the formation of spherical microdomains with localized higher growth factor concentrations. Release profiles, determined with ELISA, were linked to the structural changes that were monitored. Distinct, controlled release profiles were achieved in all formulations and showed that PEG is a versatile tool in the effective control of release rates from microspheres. Higher PEG/PLGA ratios in the polymer were shown to increase the release rate from the microspheres. In contrast, PEG administered to the inner phase decreased the release rate. The biological activity of released protein was shown in vitro in an alkaline phosphatase assay. It was demonstrated that PEG-PLGA microspheres are a promising sustained delivery system which allows a reduction of the required rhBMP-2 dose to limit both adverse effects and costs. Furthermore, the data indicated that the use of PEG as an inner phase cosolvent is not suitable for rhBMP-2 in contrast the reported beneficial effects for other growth factors.

Characterization of scaffolds for tissue engineering by benchtop-magnetic resonance imaging.

Several publications have shown approaches for the optimization of tissue engineering constructs by magnetic resonance imaging (MRI). However, the technology is still scarcely used, probably because of the poor spatial resolution of clinical scanners and their temporally limited availability for many researchers. The new benchtop- MRI (BT-MRI) equipment used in the present study is much more affordable, for example, because of the low static magnetic field strength of 0.5 T and the absence of a helium cooling system. In this study, the method of BT-MRI was evaluated for the characterization of a tissue engineering scaffold. Hollow cylinder scaffolds were made of hydroxyapatite (HA), collagen, and chitosan and wrapped in a polyglycolic acid mesh. Mass transport between construct and surrounding medium was investigated by dynamic contrast agent-enhanced MRI with gadolinium(III)-diethylaminepentaacetic acid. The results demonstrate that BT-MRI permits detailed, space-resolved insights into diffusion processes within the three-dimensional matrices, enabling a comparison of the mass transport inside different scaffold types. Inhomogeneities of the HA distribution in scaffolds caused by the fabrication were also visible in MR images. The fate of cells, labeled with superparamagnetic iron oxide nanoparticles and seeded on the scaffold surface, was monitored. For the first time, it was shown that mass transport, inhomogeneities of the HA distribution, and localization of superparamagnetic iron oxide nanoparticle-labeled cells are accessible in a tissue engineering scaffold by BT-MRI. Hence, it is demonstrated that BT-MRI is a powerful analytic method for the noninvasive evaluation of tissue engineering constructs.