Cell Morphology on Poly(methyl methacrylate) Microstructures as Function of Surface Energy.

Whilst the significance of substrate topography as a regulator of cell function is well established, a systematic analysis of the principles underlying this is still unavailable. Here we evaluate the hypothesis that surface energy plays a decisive role in substrate-mediated modulation of cell phenotype by evaluation of cell behaviour on synthetic microstructures exhibiting pronounced differences in surface energy. These microstructures, specifically cubes and walls, were fabricated from a biocompatible base polymer, poly(methyl methacrylate), by variotherm injection molding. The dimensions of the cubes were 1 µm x 1 µm x 1 µm (height x width x length) with a periodicity of 1:1 and 1:5 and the dimensions of the walls 1 µm x 1 µm x 15 mm (height x width x length) with a periodicity of 1:1 and 1:5. Mold inserts were made by lithography and electroplating. The surface energy of the resultant microstructures was determined by static contact angle measurements. Light scanning microscopy of the morphology of NT2/D1 and MC3T3-E1 preosteoblast cells cultured on structured PMMA samples in both cases revealed a profound surface energy dependence. "Walls" appeared to promote significant cell elongation, whilst a lack of cell adhesion was observed on "cubes" with the lowest periodicity. Contact angle measurements on walls revealed enhanced surface energy anisotropy (55 mN/m max., 10 mN/m min.) causing a lengthwise spreading of the test liquid droplet, similar to cell elongation. Surface energy measurements for cubes revealed increased isotropic hydrophobicity (87° max., H2O). A critical water contact angle of ≤ 80° appears to be necessary for adequate cell adhesion. A "switch" for cell adhesion and subsequently cell growth could therefore be applied by, for example, adjusting the periodicity of hydrophobic structures. In summary cell elongation on walls and a critical surface energy level for cell adhesion could be produced for NT2/D1 and MC3T3-E1 cells by symmetrical and asymmetrical energy barrier levels. We, furthermore, propose a water-drop model providing a common physicochemical cause regarding similar cell/droplet geometries and cell adhesion on the investigated microstructures.

Polymer-based metal nano-coated disposable target for matrix-assisted and matrix-free laser desorption/ionization mass spectrometry.

The ideal MALDI/LDI mass spectrometry sample target for an axial TOF instrument possesses a variety of properties. Primarily, it should be chemically inert to the sample, i.e. analyte, matrix and solvents, highly planar across the whole target, without any previous chemical contact and provide a uniform surface to facilitate reproducible measurements without artifacts from previous sample or matrix compounds. This can be hard to achieve with a metal target, which has to be extensively cleaned every time after use. Any cleaning step may leave residues behind, may change the surface properties due to the type of cleaning method used or even cause microscopic scratches over time hence altering matrix crystallization behavior. Alternatively, use of disposable targets avoids these problems. As each possesses the same surface they therefore have the potential to replace the conventional full metal targets so commonly employed. Furthermore, low cost single-use targets with high planarity promise an easier compliance with GLP guidelines as they alleviate the problem of low reproducibility due to inconsistent sample/matrix crystallization and changes to the target surface properties. In our tests, polymeric metal nano-coated targets were compared to a stainless steel reference. The polymeric metal nano-coated targets exhibited all the performance characteristics for a MALDI MS sample support, and even surpassed the - in our lab commonly used - reference in some aspects like limit of detection. The target exhibits all necessary features such as electrical conductivity, vacuum, laser and solvent compatibility.

Effects of the polymeric niche on neural stem cell characteristics during primary culturing.

The polymeric niche encountered by cells during primary culturing can affect cell fate. However, most cell types are primarily propagated on polystyrene (PS). A cell type specific screening for optimal primary culture polymers particularly for regenerative approaches seems inevitable. The effect of physical and chemical properties of treated (corona, oxygen/nitrogen plasma) and untreated cyclic olefin polymer (COP), polymethymethacrylate (PMMA), PP, PLA, PS, PC on neuronal stem cell characteristics was analyzed. Our comprehensive approach revealed plasma treated COP and PMMA as optimal polymers for primary neuronal stem cell culturing and propagation. An increase in the number of NT2/D1 cells with pronounced adhesion, metabolic activities and augmented expression of neural precursor markers was associated to the plasma treatment of surfaces of COP and PMMA with nitrogen or oxygen, respectively. A shift towards large cell sizes at stable surface area/volume ratios that might promote the observed increase in metabolic activities and distinct modulations in F-actin arrangements seem to be primarily mediated by the plasma treatment of surfaces. These results indicate that the polymeric niche has a distinct impact on various cell characteristics. The selection of distinct polymers and the controlled design of an optimized polymer microenvironment might thereby be an effective tool to promote essential cell characteristics for subsequent approaches.

Intact cell/intact spore mass spectrometry (IC/ISMS) on polymer-based, nano-coated disposable targets.

Identification and differentiation of microorganisms has and still is a long arduous task, involving culturing of the organism in question on different growth media. This procedure, which is still commonly applied, is an established method, but takes a lot of time, up to several days or even longer. It has thus been a great achievement when other analytical tools like matrix-assisted laser desorption/ionization (MALDI) mass spectrometry were introduced for faster analysis based on the surface protein pattern. Differentiation and identification of human pathogens as well as plant/animal pathogens is of increasing importance in medical care (e.g. infection, sepsis, and antibiotics resistance), biotechnology, food sciences and detection of biological warfare agents. A distinction between microorganisms on the species and strain level was made by comparing peptide/protein profiles to patterns already stored in databases. These profiles and patterns were obtained from the surface of vegetative forms of microorganisms or even their spores by MALDI MS. Thus, an unknown sample can be compared against a database of known pathogens or microorganisms of interest. To benefit from newly available, metal-based disposable microscope-slide format MALDI targets that promise a clean and even surface at a fraction of the cost from full metal targets or MTP (microtiter plate) format targets, IC/ISMS analysis was performed on these and the data evaluated. Various types of bacteria as well as fungal spores were identified unambiguously on this disposable new type of metal nano-coated targets. The method even allowed differentiation between strains of the same species. The results were compared with those gained from using full metal standard targets and found to be equal or even better in several aspects, making the use of disposable MALDI targets a viable option for use in IC/ISMS, especially e.g. for large sample throughput and highly pathogenic species.