Benjamin Richter

Two‐Component Polymer Scaffolds for Controlled Three‐Dimensional Cell Culture

O N Cell behavior is governed by interactions with the cellular environment. [ 1–3 ] These interactions include cell–cell as well as cell–extracellular matrix (ECM) contacts and act, in addition to soluble growth factors, as key regulators of cell survival, proliferation, and differentiation. However, not only the molecular composition of the contact sites, but also their spatial distributions impact cell behavior. [ 4 , 5 ] Realizing cell-culture scaffolds that mirror the complex in vivo arrangement of ECM components is an active area of biomaterials engineering. Corresponding 2D lithographically defi ned micropatterned structures are widely used and have even become commercially available. [ 6–9 ] In addition to patterned ligand distributions, mechanical interactions between cells and their environment also play an important role in regulating cellular functions. [ 10 ] Consequently, corresponding fl exible substrates were developed that allow measurements of cellular forces in 2D on planar substrates or pillar arrays. [ 11 , 12 ]

Micro-engineered 3D scaffolds for cell culture studies.

Cells in physiological 3D environments differ considerably in morphology and differentiation from those in 2D tissue culture. Naturally derived polymer systems are frequently used to study cells in 3D. These 3D matrices are complex with respect to their chemical composition, mechanical properties, and geometry. Therefore, there is a demand for well-defined 3D scaffolds to systematically investigate cell behavior in 3D. Here, fabrication techniques, materials, architectures, biochemical functionalizations, and mechanical properties of 3D scaffolds are discussed. In particular, work focusing on single cells and small cell assemblies grown in tailored synthetic 3D scaffolds fabricated by computer-based techniques are reviewed and the influence of these environments on cell behavior is evaluated.

Three‐Dimensional Microscaffolds Exhibiting Spatially Resolved Surface Chemistry

Spatial control over the surface chemistry of 3D organic-inorganic hybrid microscaffolds is achieved by a two-photon-triggered cycloaddition. Following a coating step with photoactivatable dienes via silanization, surface irradiation with a femtosecond-pulsed laser in the presence of functional dienophiles enables a site-selective alteration of the surface chemistry. Bioconjugation with fluorescent protein tags is employed to reveal the 3D molecular patterns.

Preparation of Reactive Three-Dimensional Microstructures via Direct Laser Writing and Thiol-ene Chemistry

Three-dimensional microstructures are fabricated employing the direct laser writing process and radical thiol-ene polymerization. The resin system consists of a two-photon photoinitiator and multifunctional thiols and olefins. Woodpile photonic crystals with 22 layers and a rod distance of 2 μm are fabricated. The structures are characterized via scanning electron microscopy and focused ion beam milling. The thiol-ene polymerization during fabrication is verified via infrared spectroscopy. The structures are grafted in a subsequent thiol-Michael addition reaction with different functional maleimides. The success of the grafting reaction is evaluated via laser scanning microscopy and X-ray photoelectron spectroscopy. The grafting density is calculated to be close to 200 molecules μm(-2) .

On-chip microlasers for biomolecular detection via highly localized deposition of a multifunctional phospholipid ink

We report on a novel approach to realize on-chip microlasers, by applying highly localized and material-saving surface functionalization of passive photonic whispering gallery mode microresonators. We apply dip-pen nanolithography on a true three-dimensional structure. We coat solely the light-guiding circumference of pre-fabricated poly(methyl methacrylate) resonators with a multifunctional molecular ink. The functionalization is performed in one single fabrication step and simultaneously provides optical gain as well as molecular binding selectivity. This allows for a direct and flexible realization of on-chip microlasers, which can be utilized as biosensors in optofluidic lab-on-a-chip applications. In a proof-of-concept we show how this highly localized molecule deposition suffices for low-threshold lasing in air and water, and demonstrate the capability of the ink-lasers as biosensors in a biotin-streptavidin binding experiment.

Guiding Cell Attachment in 3D Microscaffolds Selectively Functionalized with Two Distinct Adhesion Proteins.

The combination of three different photoresists into a single direct laser written 3D microscaffold permits functionalization with two bioactive full-length proteins. The cell-instructive microscaffolds consist of a passivating framework equipped with light activatable constituents featuring distinct protein-binding properties. This allows directed cell attachment of epithelial or fibroblast cells in 3D.

Cell type-specific adaptation of cellular and nuclear volume in micro-engineered 3D environments.

Bio-functionalized three-dimensional (3D) structures fabricated by direct laser writing (DLW) are structurally and mechanically well-defined and ideal for systematically investigating the influence of three-dimensionality and substrate stiffness on cell behavior. Here, we show that different fibroblast-like and epithelial cell lines maintain normal proliferation rates and form functional cell-matrix contacts in DLW-fabricated 3D scaffolds of different mechanics and geometry. Furthermore, the molecular composition of cell-matrix contacts forming in these 3D micro-environments and under conventional 2D culture conditions is identical, based on the analysis of several marker proteins (paxillin, phospho-paxillin, phospho-focal adhesion kinase, vinculin, β1-integrin). However, fibroblast-like and epithelial cells differ markedly in the way they adapt their total cell and nuclear volumes in 3D environments. While fibroblast-like cell lines display significantly increased cell and nuclear volumes in 3D substrates compared to 2D substrates, epithelial cells retain similar cell and nuclear volumes in 2D and 3D environments. Despite differential cell volume regulation between fibroblasts and epithelial cells in 3D environments, the nucleus-to-cell (N/C) volume ratios remain constant for all cell types and culture conditions. Thus, changes in cell and nuclear volume during the transition from 2D to 3D environments are strongly cell type-dependent, but independent of scaffold stiffness, while cells maintain the N/C ratio regardless of culture conditions.

3D Laser Micro- and Nano-Printing: Challenges for Chemistry

3D printing is a powerful emerging technology for the tailored fabrication of advanced functional materials. This Review summarizes the state-of-the art with regard to 3D laser micro- and nanoprinting and explores the chemical challenges limiting its full exploitation: from the development of advanced functional materials for applications in cell biology and electronics to the chemical barriers that need to be overcome to enable fast writing velocities with resolution below the diffraction limit. We further explore chemical means to enable direct laser writing of multiple materials in one resist by highly wavelength selective (λ-orthogonal) photochemical processes. Finally, chemical processes to construct adaptive 3D written structures that are able to respond to external stimuli, such as light, heat, pH value, or specific molecules, are highlighted, and advanced concepts for degradable scaffolds are explored.

Tunable free space optical delay interferometer for demodulation of differential phase shift keying signals

With a continuously tunable free space optical delay interferometer and direct detection D(Q)PSK signals can be received at varying symbol rates or under optimized conditions. The interferometers phase is stabilized and its polarization dependence mitigated.

Photonic molecules with a tunable inter-cavity gap

Optical micro-resonators have broad applications. They are used, for example, to enhance light–matter interactions in optical sensors or as model systems for investigating fundamental physical mechanisms in cavity quantum electrodynamics. Coupling two or more micro-cavities is particularly interesting as it enlarges the design freedom and the field of application. In this context, achieving tunability of the coupling strength and hence the inter-cavity gap is of utmost importance for adjusting the properties of the coupled micro-resonator system. In this paper, we report on a novel coupling approach that allows highly precise tuning of the coupling gap of polymeric micro-resonators that are fabricated side by side on a common substrate. We structure goblet-shaped whispering-gallery-mode resonators on an elastic silicone-based polymer substrate by direct laser writing. The silicone substrate is mechanically stretched in order to exploit the lateral shrinkage to reduce the coupling gap. Incorporating a laser dye into the micro-resonators transforms the cavities into micro-lasers that can be pumped optically. We have investigated the lasing emission by micro-photoluminescence spectroscopy, focusing on the spatial localization of the modes. Our results demonstrate the formation of photonic molecules consisting of two or even three resonators, for which the coupling strengths and hence the lasing performance can be precisely tuned. Flexibility and tunability are key elements in future photonics, making our approach interesting for various photonic applications. For instance, as our coupling approach can also be extended to larger cavity arrays, it might serve as a platform for tunable coupled-resonator optical waveguide devices.