Composite material consisting of microporous ß-TCP ceramic and alginate for delayed release of antibiotics.

OBJECTIVE: The aim of this study was to produce a novel composite of microporous ß-TCP filled with alginate and Vancomycin (VAN) to prolong the release behavior of the antibiotic for up to 28days. MATERIAL AND METHODS: Using the flow chamber developed by the group, porous ceramics in a directional flow were filled with alginates of different composition containing 50mg/mL of antibiotics. After cross-linking the alginate with calcium ions, incubation took place in 10mL double-distilled water for 4weeks at 37°C. At defined times (1, 2, 3, 6, 9, 14, 20 and 28days), the liquid was completely exchanged and analyzed by capillary zone electrophoresis and microtiter trials. For statistical purposes, the mean and standard deviation were calculated and analyzed by ANOVA. RESULTS: The release of VAN from alginate was carried out via an external calcium source over the entire period with concentrations above the minimal inhibitory concentration (MIC). The burst release measured 35.2±1.5%. The release of VAN from alginate with an internal calcium source could only be observed over 14days. The burst release here was 61.9±4.3%. The native alginate's burst release was 54.1±7.8%; that of the sterile alginate 40.5±6.4%. The microtiter experiments revealed efficacy over the entire study period for VAN. The MIC value was determined in the release experiments as well in a range of 0.5-2.0µg/mL against Staphylococcus aureus. STATEMENT OF SIGNIFICANCE: Drug release systems based on ß-TCP and hydrogels are well documented in literature. However, in all described systems the ceramic, as granule or powder, is inserted into a hydrogel. In our work, we do the opposite, a hydrogel which acts as reservoir for antibiotics is placed into a porous biodegradable ceramic. Eventually, this system should be applied as treatment of bone infections. Contrary to the "granule in hydrogel" composites it has the advantage of mechanical stability. Thus, it can take over functions of the bone during the healing process. For a quicker translation from our scientific research into clinical use, only FDA approved materials were used in this work.

Novel Method for Loading Microporous Ceramics Bone Grafts by Using a Directional Flow.

The aim of this study was the development of a process for filling the pores of a ß-tricalcium phosphate ceramic with interconnected porosity with an alginate hydrogel. For filling of the ceramics, solutions of alginate hydrogel precursors with suitable viscosity were chosen as determined by rheometry. For loading of the porous ceramics with the gel the samples were placed at the flow chamber and sealed with silicone seals. By using a vacuum induced directional flow, the samples were loaded with alginate solutions. The loading success was controlled by ESEM and fluorescence imaging using a fluorescent dye (FITC) for staining of the gel. After loading of the pores, the alginate is transformed into a hydrogel through crosslinking with CaCl2 solution. The biocompatibility of the obtained composite material was tested with a live dead cell staining by using MG-63 Cells. The loading procedure via vacuum assisted directional flow allowed complete filling of the pores of the ceramics within a few minutes (10 ± 3 min) while loading through simple immersion into the polymer solution or through a conventional vacuum method only gave incomplete filling.

Universal nucleic acid sequence-based amplification for simultaneous amplification of messengerRNAs and microRNAs.

A universal NASBA assay is presented for simultaneous amplification of multiple microRNA (miRNA) and messengerRNA (mRNA) sequences. First, miRNA and mRNA sequences are reverse transcribed using tailed reverse transcription primer pairs containing a gene-specific and an non-specific region. For reverse transcription of small miRNA molecules a non-specific region is incorporated into a structured stem-loop reverse transcription primer. Second, a universal NASBA primer pair that recognizes the tagged cDNA molecules enables a simultaneous, transcription-based amplification reaction (NASBA) of all different cDNA molecules in one reaction. The NASBA products (RNA copies) are detected by gene-specific DNA probes immobilized on a biochip. By using the multiplex reverse transcription combined with the universal NASBA amplification up to 14 different mRNA and miRNA sequences can be specifically amplified and detected in parallel. In comparison with standard multiplex NASBA assays this approach strongly enhances the multiplex capacity of NASBA-based amplification reactions. Furthermore simultaneous assaying of different RNA classes can be achieved that might be beneficial for studying miRNA-based regulation of gene expression or for RNA-based tumor diagnostics.