Matching Together Living Cells and Prototissues: Will There Be Chemistry?

Scientific advancements in bottom-up synthetic biology have led to the development of numerous models of synthetic cells, or protocells. To date, research has mainly focused on increasing the (bio)chemical complexity of these bioinspired micro-compartmentalized systems, yet the successful integration of protocells with living cells remains one of the major challenges in bottom-up synthetic biology. In this review, we aim to summarize the current state of the art in hybrid protocell/living cell and prototissue/living cell systems. Inspired by recent breakthroughs in tissue engineering, we review the chemical, bio-chemical, and mechano-chemical aspects that hold promise for achieving an effective integration of non-living and living matter. The future production of fully integrated protocell/living cell systems and increasingly complex prototissue/living tissue systems not only has the potential to revolutionize the field of tissue engineering, but also paves the way for new technologies in (bio)sensing, personalized therapy, and drug delivery.

Site Selectivity of Peptoids as Azobenzene Scaffold for Molecular Solar Thermal Energy Storage.

Storing solar energy is a key challenge in modern science. MOlecular Solar Thermal (MOST) systems, in particular those based on azobenzene switches, have received great interest in the last decades. The energy storage properties of azobenzene (t1/2 <4 days; ΔH~270 kJ/kg) must be improved for future applications. Herein, we introduce peptoids as programmable supramolecular scaffolds to improve the energy storage properties of azobenzene-based MOST systems. We demonstrate with 3-unit peptoids bearing a single azobenzene chromophore that dynamics of the MOST systems can be tuned depending on the anchoring position of the photochromic unit on the macromolecular backbone. We measured a remarkable increase of the half-life of the metastable form up to 14 days at 20 °C for a specific anchoring site, significantly higher than the isolated azobenzene moiety, thus opening new perspectives for MOST development. We also highlight that liquid chromatography coupled to mass spectrometry does not only enable to monitor the different stereoisomers during the photoisomerization process as traditionally done, but also allows to determine the thermal back-isomerization kinetics.

Automated analysis of soft material microindentation.

An understanding of the mechanical properties of soft hydrogel materials over multiple length scales is important for their application in many fields. Typical measurement methods provide either bulk mechanical properties (compression, tensile, rheology) or probing of nano or microscale properties and heterogeneity (nanoindentation, AFM). In this work we demonstrate the complementarity of instrumented microindentation to these techniques, as it provides representative Young's moduli for soft materials with minimal influence of the experimental parameters chosen, and allows mechanical property mapping across macroscopic areas. To enable automated analysis of the large quantities of data required for these measurements, we develop a new fitting algorithm to process indentation data. This method allows for the determination of Young's moduli from imperfect data by automatic selection of a region of the indentation curve which does not display inelastic deformation or substrate effects. We demonstrate the applicability of our approach with a range of hydrogels, including materials with patterns and gradients in stiffness, and expect the techniques described here to be useful developments for the mechanical analysis of a wide range of soft and biological systems.

Asymmetric Dressing of WSe2 with (Macro)molecular Switches: Fabrication of Quaternary-Responsive Transistors.

The forthcoming saturation of Moore's law has led to a strong demand for integrating analogue functionalities within semiconductor-based devices. As a step toward this goal, we fabricate quaternary-responsive WSe2-based field-effect transistors (FETs) whose output current can be remotely and reversibly controlled by light, heat, and electric field. A photochromic silane-terminated spiropyran (SP) is chemisorbed on SiO2 forming a self-assembled monolayer (SAM) that can switch from the SP to the merocyanine (MC) form in response to UV illumination and switch back by either heat or visible illumination. Such a SAM is incorporated at the dielectric-semiconductor interface in WSe2-based FETs. Upon UV irradiation, a drastic decrease in the output current of 82% is observed and ascribed to the zwitterionic MC isomer acting as charge scattering site. To provide an additional functionality, the WSe2 top surface is coated with a ferroelectric co-polymer layer based on poly(vinylidene fluoride-co-trifluoroethylene). Because of its switchable inherent electrical polarization, it can promote either the accumulation or depletion of charge carriers in the WSe2 channel, thereby inducing a current modulation with 99% efficiency. Thanks to the efficient tuning induced by the two components and their synergistic effects, the device polarity could be modulated from n-type to p-type. Such a control over the carrier concentration and device polarity is key to develop 2D advanced electronics. Moreover, the integration strategy of multiple stimuli-responsive elements into a single FET allows us to greatly enrich its functionality, thereby promoting the development for More-than-Moore technology.

A Floating Mold Technique for the Programmed Assembly of Protocells into Protocellular Materials Capable of Non-Equilibrium Biochemical Sensing.

Despite important breakthroughs in bottom-up synthetic biology, a major challenge still remains the construction of free-standing, macroscopic, and robust materials from protocell building blocks that are stable in water and capable of emergent behaviors. Herein, a new floating mold technique for the fabrication of millimeter- to centimeter-sized protocellular materials (PCMs) of any shape that overcomes most of the current challenges in prototissue engineering is reported. Significantly, this technique also allows for the generation of 2D periodic arrays of PCMs that display an emergent non-equilibrium spatiotemporal sensing behavior. These arrays are capable of collectively translating the information provided by the external environment and are encoded in the form of propagating reaction-diffusion fronts into a readable dynamic signal output. Overall, the methodology opens up a route to the fabrication of macroscopic and robust tissue-like materials with emergent behaviors, providing a new paradigm of bottom-up synthetic biology and biomimetic materials science.

Bioinspired Networks of Communicating Synthetic Protocells.

The bottom-up synthesis of cell-like entities or protocells from inanimate molecules and materials is one of the grand challenges of our time. In the past decade, researchers in the emerging field of bottom-up synthetic biology have developed different protocell models and engineered them to mimic one or more abilities of biological cells, such as information transcription and translation, adhesion, and enzyme-mediated metabolism. Whilst thus far efforts have focused on increasing the biochemical complexity of individual protocells, an emerging challenge in bottom-up synthetic biology is the development of networks of communicating synthetic protocells. The possibility of engineering multi-protocellular systems capable of sending and receiving chemical signals to trigger individual or collective programmed cell-like behaviours or for communicating with living cells and tissues would lead to major scientific breakthroughs with important applications in biotechnology, tissue engineering and regenerative medicine. This mini-review will discuss this new, emerging area of bottom-up synthetic biology and will introduce three types of bioinspired networks of communicating synthetic protocells that have recently emerged.

Graphene transistors for real-time monitoring molecular self-assembly dynamics.

Mastering the dynamics of molecular assembly on surfaces enables the engineering of predictable structural motifs to bestow programmable properties upon target substrates. Yet, monitoring self-assembly in real time on technologically relevant interfaces between a substrate and a solution is challenging, due to experimental complexity of disentangling interfacial from bulk phenomena. Here, we show that graphene devices can be used as highly sensitive detectors to read out the dynamics of molecular self-assembly at the solid/liquid interface in-situ. Irradiation of a photochromic molecule is used to trigger the formation of a metastable self-assembled adlayer on graphene and the dynamics of this process are monitored by tracking the current in the device over time. In perspective, the electrical readout in graphene devices is a diagnostic and highly sensitive means to resolve molecular ensemble dynamics occurring down to the nanosecond time scale, thereby providing a practical and powerful tool to investigate molecular self-organization in 2D.

A New Class of Rigid Multi(azobenzene) Switches Featuring Electronic Decoupling: Unravelling the Isomerization in Individual Photochromes.

We report a novel class of star-shaped multiazobenzene photoswitches comprising individual photochromes connected to a central trisubstituted 1,3,5-benzene core. The unique design of such C3-symmetric molecules, consisting of conformationally rigid and pseudoplanar scaffolds, made it possible to explore the role of electronic decoupling in the isomerization of the individual azobenzene units. The design of our tris-, bis-, and mono(azobenzene) compounds limits the π-conjugation between the switches belonging to the same molecule, thus enabling the efficient and independent isomerization of each photochrome. An in-depth experimental insight by making use of different complementary techniques such as UV-vis absorption spectroscopy, high performance liquid chromatography, and advanced mass spectrometry methods as ion mobility revealed an almost complete absence of electronic delocalization. Such evidence was further supported by both experimental (electrochemistry, kinetical analysis) and theoretical (DFT calculations) analyses. The electronic decoupling provided by this molecular design guarantees a remarkably efficient photoswitching of all azobenzenes, as evidenced by their photoisomerization quantum yields, as well as by the Z-rich UV photostationary states. Ion mobility mass spectrometry was exploited for the first time to study multiphotochromic compounds revealing the occurrence of a large molecular shape change in such rigid star-shaped azobenzene derivatives. In view of their high structural rigidity and efficient isomerization, our multiazobenzene photoswitches can be used as key components for the fabrication of complex stimuli-responsive porous materials.

Electronic Decoupling in C3-Symmetrical Light-Responsive Tris(Azobenzene) Scaffolds: Self-Assembly and Multiphotochromism.

We report the synthesis of a novel C3-symmetrical multiphotochromic molecule bearing three azobenzene units at positions 1, 3, 5 of the central phenyl ring. The unique geometrical design of such a rigid scaffold enables the electronic decoupling of the azobenzene moieties to guarantee their simultaneous isomerization. Photoswitching of all azobenzenes in solution was demonstrated by means of UV-vis absorption spectroscopy and high performance liquid chromatography (HPLC) analysis. Scanning tunneling microscopy investigations at the solid-liquid interface, corroborated by molecular modeling, made it possible to unravel the dynamic self-assembly of such systems into ordered supramolecular architectures, by visualizing and identifying the patterns resulting from three different isomers, thereby demonstrating that the multiphotochromism is retained when the molecules are confined in two dimensions.

Fluorescence Commutation and Surface Photopatterning with Porphyrin Tetradithienylethene Switches.

Four tetradithienylethene (DTE) substituted porphyrins, the free base 1H2 , and three metal derivatives (1Zn, 1Co, 1Ni), were synthesized and studied. These dyads, for which the DTE units are connected to the porphyrin's meso positions via a meta-phenyl spacer, exhibit reversible photochromic properties in all cases, with conversion to the photostationary state (PSS) up to 88 %, as confirmed by absorption and NMR spectroscopies. Compounds 1H2 and 1Zn are fluorescent in solution and display a red emission. Upon irradiation with UV light to trigger the closing of the DTEs, the fluorescence of both the free base and zinc porphyrin was very efficiently quenched in solution. The reversible, photo-switching of luminescence was retained in a tetra-DTE free-base porphyrin-doped polystyrene film, for which photo-patterning was demonstrated by confocal scanning microscopy. The tunable fluorescent properties of this multi-DTE framework render this compound of interest as a photo-rewritable fluorescent ink.

Photoisomerisation and light-induced morphological switching of a polyoxometalate-azobenzene hybrid.

The functionalization of a spherical Keplerate-type polyoxometalate {Mo72V30} with a cationic azobenzene surfactant has been achieved through ionic self-assembly. The photoisomerisation reaction of this complex, which emerges in a light-triggered aggregation-disaggregation process, has been followed by 1H NMR spectroscopy, dynamic light scattering, absorption spectroscopy and scanning electron microscopy analyses.

Exploring the Effect of Ligand Structural Isomerism in Langmuir-Blodgett Films of Chiral Luminescent Eu(III) Self-Assemblies.

Here we have investigated the influence of the antenna group position on both the formation of chiral amphiphilic Eu(III) -based self-assemblies in CH3 CN solution and, on the ability to form monolayers on the surface of quartz substrates using the Langmuir-Blodgett technique, by changing from the 1-naphthyl (2(R), 2(S)) to the 2-naphthyl (1(R), 1(S)) position. The evaluation of binding constants of the self- assemblies in CH3 CN solution was achieved using conventional techniques such as UV/Visible and luminescence spectroscopies along with more specific circular dichroism (CD) spectroscopy. The binding constants obtained for EuL, EuL2 and EuL3 species in the case of 2-naphthyl derivatives were comparable to those obtained for 1-naphthyl derivatives. The analysis of the changes in the CD spectra of 1(R) and 1(S) upon addition of Eu(III) not only allowed us to evaluate the values of the binding constants but the resulting recalculated spectra may also be used as fingerprints for assignment of the chiral self-assembly species formed in solution. The obtained monolayers were predominantly formed from EuL3 (≈85 %) with the minor species present in ≈15 % EuL2 .