Sabine Illner

Industrial biotechnology—the future of green chemistry?

Biocatalysis fulfils many key criteria of green chemistry, but this raises the question: how efficient are such production methods and how important is biotechnology for greener industrial chemistry today? This review summarizes the advantages, disadvantages and potential uses of biocatalysis to undertake greener chemistry. By addressing the obstacles of biocatalytic approaches, it will be demonstrated that continuous improvement is required to overcome existing limitations. To illustrate the state of the art in the use of enzymes and microorganisms, representative examples of industrial chemistry are discussed and evaluated in this article. Several biocatalytic processes are compared to their chemical alternatives. Finally, future trends for the biocatalytic reductions of amides and the asymmetric hydrogenation of olefins are discussed.

A Falling-Film Microreactor for Enzymatic Oxidation of Glucose

Many oxidation processes require the presence of molecular oxygen in the reaction media. Reactors are needed that provide favorable conditions for the mass transfer between the gas and the liquid phase. In this study, two recent key technologies, microreactor technology and biotechnology, were combined to present an interesting alternative to conventional methods and open up excellent possibilities to intensify chemical processes in the field of fine chemicals. An enzyme‐catalyzed gas/liquid phase reaction in a falling‐film microreactor (FFMR) was examined for the first time. The test reaction was the oxidation of β‐D‐glucose to gluconic acid catalyzed by glucose oxidase (GOx). Various factors influencing the biotransformation, such as oxygen supply, temperature, enzyme concentration, and reaction time were investigated and compared to those in conventional batch systems. The most critical factor, the volumetric mass‐transfer coefficient for the efficient use of oxygen‐dependent enzymes, was determined by using the integrated online detection of dissolved oxygen in all systems. The extremely large surface‐to‐volume ratio of the FFMR facilitated the contact between the enzyme solution and the gaseous substrate. Hence, in a continuous bubble‐free FFMR system with a residence time of 25 seconds, a final conversion of up to 50 % in enzymatic oxidation was reached, whereas conversion in a conventional bubble column resulted in only 27 %. Finally, an option for scale‐up was shown through an enlarged version of the FFMR.

Surface Modification of Biodegradable Polymers towards Better Biocompatibility and Lower Thrombogenicity.

Purpose Drug-eluting stents (DES) based on permanent polymeric coating matrices have been introduced to overcome the in stent restenosis associated with bare metal stents (BMS). A further step was the development of DES with biodegradable polymeric coatings to address the risk of thrombosis associated with first-generation DES. In this study we evaluate the biocompatibility of biodegradable polymer materials for their potential use as coating matrices for DES or as materials for fully bioabsorbable vascular stents. Materials and Methods Five different polymers, poly(L-lactide) PLLA, poly(D,L-lactide) PDLLA, poly(L-lactide-co-glycolide) P(LLA-co-GA), poly(D,L-lactide-co-glycolide) P(DLLA-co-GA) and poly(L-lactide-co-ε-caprolactone), P(LLA-co-CL) were examined in vitro without and with surface modification. The surface modification of polymers was performed by means of wet-chemical (NaOH and ethylenediamine (EDA)) and plasma-chemical (O2 and NH3) processes. The biocompatibility studies were performed on three different cell types: immortalized mouse fibroblasts (cell line L929), human coronary artery endothelial cells (HCAEC) and human umbilical vein endothelial cells (HUVEC). The biocompatibility was examined quantitatively using in vitro cytotoxicity assay. Cells were investigated immunocytochemically for expression of specific markers, and morphology was visualized using confocal laser scanning (CLSM) and scanning electron (SEM) microscopy. Additionally, polymer surfaces were examined for their thrombogenicity using an established hemocompatibility test. Results Both endothelial cell types exhibited poor viability and adhesion on all five unmodified polymer surfaces. The biocompatibility of the polymers could be influenced positively by surface modifications. In particular, a reproducible effect was observed for NH3-plasma treatment, which enhanced the cell viability, adhesion and morphology on all five polymeric surfaces. Conclusion Surface modification of polymers can provide a useful approach to enhance their biocompatibility. For clinical application, attempts should be made to stabilize the plasma modification and use it for coupling of biomolecules to accelerate the re-endothelialization of stent surfaces in vivo.

In vitro study of sirolimus release from nonwoven PLLA matrices

Abstract Sirolimus incorporated nonwoven polymer matrices were fabricated via electrospinning. Release kinetics considering different fiber diameters and layer thicknesses were investigated. In vitro drug release profiles were evaluated by measuring the drug concentration in an established drug release medium (0.9% saline solution with additives, not buffered) at predetermined time points. Furthermore, an NH3-plasma pretreatment was examined to ensure complete wetting from the beginning of the study. In comparison to thin drug-loaded PLLA spray coatings it was shown that the release of sirolimus is diffusion- and degradation-controlled regardless of the surface-to-volume ratio, though fiber diameters or a hydrophilization can affect its release kinetics.

Fast parallel quantification of dual drug systems via UV-VIS spectrometry – a case study with antibiotic substances

Abstract Implant-based drug delivery necessitates a rigorous quantitative assessment and quality control of the drug release behavior. In this context, a fast simultaneous UV-VIS spectrometer quantification method was developed for an antibiotic dual drug system. Particular challenges arose from interfering spectra, as well as different solubility and stability of the drugs. The test system used here consists of the antibiotics Minocycline and Rifampin, which are used as an antibiotic combination approach in a growing number of device applications, such as impregnation of venous or urethral catheters. Two suitable wavelengths for this test system were identified. At a wavelength of 475 nm only Rifampin shows an absorption maximum, which was used to determine the concentration of Rifampin in the mixture. At 245 nm both antibiotics have a local extremum, a maximum for Minocycline, and a minimum for Rifampin. The developed UV-VIS spectrometer method for Rifampin and Minocycline dissolved in Sörensen buffer showed good results for small concentrations of Minocycline. For stability tests, a mixture of both antibiotics (1:1) was stored at 37°C for up to 144 h in NaCl solution (0.9%), and in Sörensen buffer, respectively. At defined time points, UV-VIS spectra were recorded, and feasibility of this quick simultaneous quantification method for these two widely used antibiotics was demonstrated.

Rheological analysis of hybrid hydrogels during polymerization processes

Abstract Development of new implant coatings with temperature-controlled drug release to treat infections after device implantation can be triggered by highly elastic hydrogels with adequate stability and adhesive strength in the swollen state. By using an ionic liquid (IL [ViPrIm]+[Br]−) as additive to N-isopropylacrylamide (NIPAAm) unique effects on volumetric changes and mechanical properties as well as thermoresponsive drug release of the obtained hybrid hydrogels were observed. In this context, rheological measurements allow the monitoring of gelation processes as well as chemical, mechanical, and thermal treatments and effects of additives. Hybrid hydrogels of pNIPAAm and poly (ionic liquid) (PIL) were prepared by radical emulsion polymerization with N,N′-methylenebis(acrylamide) as 3D crosslinking agent. By varying monomer, initiator and crosslinker amounts the multi-compound system during polymerization was monitored by oscillatory time sweep experiments. The time dependence of the storage modulus (G′) and the loss modulus (G″) was measured, whereby the intersection of G′ and G″ indicates the sol-gel transition. Viscoelastic behavior and complex viscosity of crosslinked and non-crosslinked hydrogels were obtained. Within material characterization rheology can be used to determine process capability and optimal working conditions. For biomedical applications complete hydrogelation inter-connecting all compounds can be received providing the possibility to process mechanically stable, swellable implant coatings or wound closures.

Synthesis and characterization of PIL/pNIPAAm hybrid hydrogels

Abstract In this study, varying amounts of NIPAAm and an ionic liquid (IL), namely 1-vinyl-3-isopropylimidazolium bromide ([ViPrIm]+[Br]−), have been used to synthesize hybrid hydrogels by radical emulsion polymerization. Amounts of 70/30%, 50/50%, 30/70%, 15/85% and 5/95% (wt/wt) of PIL/pNIPAAm were used to produce hybrid hydrogels as well as the parental hydrogels. The adhesive strength was investigated and evaluated for mechanical characterization. Thermal properties of resulting hydrogels have been investigated using differential scanning calorimetry (DSC) in a default heating temperature range (heating rate 10 K min−1). The presence of poly ionic liquids (PIL) in the polymer matrix leads to a moved LCST (lower critical solution temperature) to a higher temperature range for certain hybrid hydrogels PIL/pNIPAAm. While pNIPAAm exhibits an LCST at 33.9 ± 0.3°C, PIL/pNIPAAm 5/95% and PIL/pNIPAAm 15/85% were found to have LCSTs at 37.6 ± 0.9°C and 52 ± 2°C, respectively. This could be used for controlled drug release that goes along with increasing body temperature in response to an implantation caused infection.