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This study aims to equip students with conceptual modeling skills to address compelling 21st-century challenges in chemistry and chemical engineering education. System-based concept mapping is a critical competence for analyzing global, often complex, problems. We examined how conceptual modeling could scaffold practical experimental design, transitioning from problem identification to testable hypotheses. We set up a project in which first-year undergraduates in chemical engineering work in groups of 5–6 students. Their task was to develop concrete hypotheses for assignments that center on finding sustainable solutions for polluted environments. A set of educational roles (i.e., lecturers, tutors, learning assistants, educational specialist, and project coordinator) were implemented to ensure that students could accomplish their main learning outcome; that is, to become familiar with the academic way of thinking and apply critical thinking skills as a team. Interviews were conducted after the project was finished and revealed that, while conceptual modeling helped students to structure their ideas (i.e., to learn how to design research questions, incorporate interventions, and test models), developing hypotheses remains a challenging task. Our findings brought us to the recommendations for teaching conceptual modeling in the curriculum rather than at the project level, allowing students to progressively transition from understanding and applying concept mapping in their first year into creating solutions within the context of solving complex real-world problems in the final year of their bachelor’s degree. The collaborative learning environment and project format employed in this work could spark new ways to teach science that facilitates systems thinking in chemistry.

Three-dimensional (3D) scaffolds with chemical diversity are significant to direct cell adhesion onto targeted surfaces, which provides solutions to further control over cell fates and even tissue formation. However, the site-specific modification of specific biomolecules to realize selective cell adhesion has been a challenge with the current methods when building 3D scaffolds. Conventional methods of immersing as-prepared structures in solutions of biomolecules lead to nonselective adsorption; recent printing methods have to address the problem of switching multiple nozzles containing different biomolecules. The recently developed concept of macroscopic supramolecular assembly (MSA) based on the idea of “modular assembly” is promising to fabricate such 3D scaffolds with advantages of flexible design and combination of diverse modules with different surface chemistry. Herein we report an MSA method to fabricate 3D ordered structures with internal chemical diversity for site-selective cell adhesion. The 3D structure is prepared via 3D alignment of polydimethylsiloxane (PDMS) building blocks with magnetic pick-and-place operation and subsequent interfacial bindings between PDMS based on host/guest molecular recognition. The site-specific cell affinity is realized by distributing targeted building blocks that are modified with polylysine molecules of opposite chiralities: PDMS modified with films containing poly-l-lysine (PLL) show higher cell density than those with poly-d-lysine (PDL). This principle of selective cell adhesion directed simply by spatial distribution of chiral molecules has been proven effective for five different cell lines. This facile MSA strategy holds promise to build complex 3D microenvironment with on-demand chemical/biological diversities, which is meaningful to study cell/material interactions and even tissue formation.

Quantification of the multivalent interactions of influenza viruses binding at interfaces may provide ways to tackle key biological questions regarding influenza virulence and zoonoses. Yet, the deconvolution of the contributions of molecular and interfacial parameters, such as valency, interaction area, and receptor density, to the binding of whole viruses is hindered by difficulties in the direct determination of these parameters. We report here a chemical platform technology to study the binding of multivalent recombinant hemagglutinin (rHA) nanoparticles at artificial sialoglycan cell receptor-presenting interfaces in which all these parameters can be derived, thus allowing the desired full and quantitative binding analysis. SiO2substrates were functionalized with supported lipid bilayers containing a targeted and tunable fraction of a biotinylated lipid, followed by the adsorption of streptavidin and biotinylated polyvalent 2,3- or 2,6-sialyl lactosamine (SLN). rHA nanoparticles were used as a virus mimic to provide a good prediction of the number of interactions involved in binding. Low nanomolar affinities and selectivities for binding at the 2,6-SLN platforms were observed for rHA particles from three different virus variants. When fitting the data to a multivalency model, the nanomolar overall affinity appears to be achieved by 6–9 HA–sugar molecular interaction pairs, which individually present a rapid association/dissociation behavior. This dynamic behavior may be an essential biological attribute in the functioning of the influenza virus.

Intrahepatic islet transplantation is a promising therapy for treatment of type 1 diabetes. During islet isolation, collagenase is used to extract islets from the pancreas, leading to loss of important cell–matrix interactions. Loss of the native pancreatic microenvironment is associated with apoptosis of islet cells, early graft failure, and poor islet function. The islet extracellular matrix (ECM) is composed of a specific combination of collagen (Col), laminin (LN), and fibronectin (FN) molecules. Reintroducing these molecules has been shown to boost the function, viability, morphology, and proliferation of β-cells. In this research, the effect of combinatorial ECM on islet function and survival was investigated. Specifically, thin-film microwell array scaffolds made from two distinct biomaterials were coated with FN, collagen type IV (Col4), LN111, LN332, or a combination thereof. We found that coatings containing a single type of ECM molecule, for example, FN or Col, can improve short-term islet function. However, these single proteins do not prevent loss of morphology and subsequent loss of islet function afterward. In contrast, combining Col4 with LN111 at a ratio of 8:2 not only improved short-term islet function but also preserved islet structure and islet function on a longer term. This effect was reproducibly shown on poly(ester-urethane) and poly(ethylene-glycol-terephthalate-poly(butylene-terephthalate) microwell islet delivery devices as well as tissue culture polystyrene. We concluded that biofunctionalization of inert biomaterials regardless of their molecular composition with a specific combination of islet ECM molecules can support and improve islet function over longer time periods. Our data suggested that creating a biomimetic islet niche by biofunctionalization of biomaterials can significantly improve the endocrine function of β-cells. The creation of islet mimicking niches in islet delivery devices leads to an improvement of islet function by restoring part of the islet's ECM in these devices.Impact StatementThis research deals with finding a proper bioengineering strategy to improve the outcome of islets transplantation for treatment of type 1 diabetes. It is focused on the mimicking of islet extracellular matrix niche in microwell islet delivery devices to improve their endocrine function.

Extracellular matrix (ECM) molecules play significant roles in regulating β-cell function and viability within pancreatic islets by providing mechanical and biological support, stimulating cell survival, proliferation, and their endocrine function. During clinical islet transplantation, the β-cell's ECM environment is degraded by enzymatic digestion. Literature suggests that interactions between islet cells and ECM molecules, such as fibronectin (FN), collagen type IV (Col4), and laminin (LN), are essential for maintaining, or stimulation of islet function and survival, and can effect differentiation and proliferation of the endocrine cells. It is also thought that three-dimensional (3D) culture of β-cells can improve glucose responsiveness by providing a specific niche, in which cells can interact with each other in a more natural manner. Conventional suspension cultures with β-cells results generally in a heterogeneous population with small and large aggregates, in which cells experience different nutrient diffusion limitations, negatively affecting their physiology and function. In this study, we have explored the effect of FN, Col4, and LN111 on INS1E insulinoma cells by using microcontact printing (μCP) to investigate whether a controlled environment and aggregate dimensions would improve their endocrine function. Using this method, we produced a pattern of well-defined circular spots of FN, Col4, and LN111 on polydimethylsiloxane with high spatial resolution. Cell seeding of the INS1E cells on these ECM protein spots resulted in the formation of 3D β-cell aggregates. We show that these INS1E aggregates have very reproducible dimensions, and that the cell culture method can be easily adjusted, leading to a highly accurate way of forming 3D β-cell aggregates on an ECM-functionalized substrate. In addition, we show that ECM molecules can act as anchoring points for β-cells on an otherwise non-cell-adherent material, and this can improve both the endocrine function and viability. We found a significant increase in the secretion of insulin by INS1E cells cultured on μCP FN and Col4 substrates, in comparison to cells that were growing in monolayers on substrates without ECM molecules. Moreover, INS1E cells growing on circular ECM spots in a 3D manner showed improved endocrine function in comparison to their two-dimensional counterparts.Impact StatementThis research deals with finding a proper bioengineering strategy for the creation of improved β-cell replacement therapy in type 1 diabetes. It specifically deals with the microenvironment of β-cells and its relationship to their endocrine function.

Release of growth factors while simultaneously maintaining their full biological activity over a period of days to weeks is an important issue in controlled drug delivery and in tissue engineering. In addition, the selected strategy to immobilize growth factors largely determines their biological activity. Silica surfaces derivatized with glycidyloxy propyl trimethoxysilane and poly(glycidyl methacrylate) brushes yielded epoxide-functionalized surfaces onto which human bone morphogenetic protein-6 (hBMP-6) was immobilized giving stable secondary amine bonds. The biological activity of hBMP-6 was unleashed by hydrolysis of the surface siloxane and ester bonds. We demonstrate that this type of labile bonding strategy can be applied to biomaterial surfaces with relatively simple and biocompatible chemistry, such as siloxane, ester, and imine bonds. Our data indicates that the use of differential hydrolytically labile linkers is a versatile method for functionalization of biomaterials with a variety of growth factors providing control over their biological activity.

We report oriented immobilization of proteins using the standard hexahistidine (His6)-Ni2+:NTA (nitrilotriacetic acid) methodology, which we systematically tuned to give control of surface coverage. Fluorescence microscopy and surface plasmon resonance measurements of self-assembled monolayers (SAMs) of red fluorescent proteins (TagRFP) showed that binding strength increased by 1 order of magnitude for each additional His6-tag on the TagRFP proteins. All TagRFP variants with His6-tags located on only one side of the barrel-shaped protein yielded a 1.5 times higher surface coverage compared to variants with His6-tags on opposite sides of the so-called β-barrel. Time-resolved fluorescence anisotropy measurements supported by polarized infrared spectroscopy verified that the orientation (and thus coverage and functionality) of proteins on surfaces can be controlled by strategic placement of a His6-tag on the protein. Molecular dynamics simulations show how the differently tagged proteins reside at the surface in “end-on” and “side-on” orientations with each His6-tag contributing to binding. Also, not every dihistidine subunit in a given His6-tag forms a full coordination bond with the Ni2+:NTA SAMs, which varied with the position of the His6-tag on the protein. At equal valency but different tag positions on the protein, differences in binding were caused by probing for Ni2+:NTA moieties and by additional electrostatic interactions between different fractions of the β-barrel structure and charged NTA moieties. Potential of mean force calculations indicate there is no specific single-protein interaction mode that provides a clear preferential surface orientation, suggesting that the experimentally measured preference for the end-on orientation is a supra-protein, not a single-protein, effect.

Biomimetic and stimuli-responsive cell-material interfaces are actively being developed to study and control various cell-dynamics phenomena. Since cells naturally reside in the highly dynamic and complex environment of the extracellular matrix, attempts are being made to replicate these conditions in synthetic biomaterials. Supramolecular chemistry, dealing with noncovalent interactions, has recently provided possibilities to incorporate such dynamicity and responsiveness in various types of architectures. Using a cucurbit[8]uril-based host–guest system, we have successfully established a dynamic and electrochemically responsive interface for the display of the integrin-specific ligand, Arg-Gly-Asp (RGD), to promote cell adhesion. Due to the weak nature of the noncovalent forces by which the components at the interface are held together, we expected that cell adhesion would also be weaker in comparison to traditional interfaces where ligands are usually immobilized by covalent linkages. To assess the stability and limitations of our noncovalent interfaces, we performed single-cell force spectroscopy studies using fluid force microscopy. This technique enabled us to measure rupture forces of multiple cells that were allowed to adhere for several hours on individual substrates. We found that the rupture forces of cells adhered to both the noncovalent and covalent interfaces were nearly identical for up to several hours. We have analyzed and elucidated the reasons behind this result as a combination of factors including the weak rupture force between linear Arg-Gly-Asp and integrin, high surface density of the ligand, and increase in effective concentration of the supramolecular components under spread cells. These characteristics enable the construction of highly dynamic biointerfaces without compromising cell-adhesive properties.

Supramolecular assemblies, formed through noncovalent interactions, has become particularly attractive to develop dynamic and responsive architectures to address living systems at the nanoscale. Cucurbit[8]uril (CB[8]), a pumpkin shaped macrocylic host molecule, has been successfully used to construct various self-assembled architectures for biomedical applications since it can simultaneously bind two aromatic guest molecules within its cavity. Such architectures can also be designed to respond to external stimuli. Integrating living organisms as an active component into such supramolecular architectures would add a new dimension to the capabilities of such systems. To achieve this, we have incorporated supramolecular functionality at the bacterial surface by genetically modifying a transmembrane protein to display a CB[8]-binding motif as part of a cystine-stabilized miniprotein. We were able to confirm that this supramolecular motif on the bacterial surface specifically binds CB[8] and forms multiple intercellular ternary complexes leading to aggregation of the bacterial solution. We performed various aggregation experiments to understand how CB[8] interacts with this bacterial strain and also demonstrate that it can be chemically reversed using a competitor. To confirm that this strain can be incorporated with a CB[8] based architecture, we show that the bacterial cells were able to adhere to CB[8] self-assembled monolayers (SAMs) on gold and still retain considerable motility for several hours, indicating that the system can potentially be used to develop supramolecular bacterial biomotors. The bacterial strain also has the potential to be combined with other CB[8] based architectures like nanoparticles, vesicles and hydrogels.

Supramolecular assembly of proteins on surfaces and vesicles was investigated by site‐selective incorporation of a supramolecular guest element on proteins. Fluorescent proteins were site‐selectively labeled with bisadamantane by SNAP‐tag technology. The assembly of the bisadamantane functionalized SNAP‐fusion proteins on cyclodextrin‐coated surfaces yielded stable monolayers. The binding of the fusion proteins is specific and occurs with an affinity in the order of 106M−1as determined by surface plasmon resonance. Reversible micropatterns of the fusion proteins on micropatterned cyclodextrin surfaces were visualized by using fluorescence microscopy. Furthermore, the guest‐functionalized proteins could be assembled out of solution specifically onto the surface of cyclodextrin vesicles. The SNAP‐tag labeling of proteins thus allows for assembly of modified proteins through a host–guest interaction on different surfaces. This provides a new strategy in fabricating protein patterns on surfaces and takes advantage of the high labeling efficiency of the SNAP‐tag with designed supramolecular elements.

Using poly(trimethylene carbonate) (PTMC) network surfaces containing unreacted macromer methacrylate groups, thiol‐functionalized biotin (SH‐undecyl‐biotin) can be bound to the surface. With micro contact printing techniques and making use of the biotin‐streptavidin (SAv) binding system patterned surfaces with biological functionality can be prepared in a very straightforward manner. This method has high potential in numerous biomedical applications.

Einfach und unter milden Bedingungenwerden Proteine durch Thiol‐En‐Photokupplung zwischen S‐Farnesylgruppen an einer genetisch kodierbaren „CAAX‐Box“‐Tetrapeptidsequenz (A ist aliphatisch) am C‐Terminus des Proteins und Oberflächen‐Thiolen immobilisiert (siehe Schema). Auf diese Art gelingt die gerichtete kovalente Immobilisierung von Proteinen direkt aus Expressionslysaten ohne zusätzliche Reinigungs‐ oder Derivatisierungsschritte.

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Progress in the development of proteinimmobilization strategies and methods has made protein biochips increasingly accessible. The integration of these assay and analysis platforms into biomedical and biotechnological research has substantially expanded the repertoire of methods available for proteomics and biomarker research and for drug development. This Minireview highlights selected developments in the application of protein biochips in these fields.

Dank Fortschritten bei Immobilisierungsstrategien für Proteine sind Proteinbiochips in den letzten Jahren immer besser verfügbar geworden. Durch die Integration dieser Assay und Analyseplattformen in die biomedizinische und biotechnologische Forschung konnte das Repertoire an Methoden für die Proteomik, die Biomarkerforschung und die Wirkstoffentwicklung deutlich vergrößert werden. In diesem Kurzaufsatz werden ausgewählte Entwicklungen bei der Anwendung von Proteinbiochips auf diesen Forschungsgebieten beleuchtet.

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Synthetically designed biomaterials strive to recapitulate and mimic the complex environment of natural systems. Using natural materials as a guide, the ability to create high performance biomaterials that control cell fate, and support the next generation of cell and tissue-based therapeutics, is starting to emerge. Supramolecular chemistry takes inspiration from the wealth of non-covalent interactions found in natural materials that are inherently complex, and using the skills of synthetic and polymer chemistry, recreates simple systems to imitate their features. Within the past decade, supramolecular biomaterials have shown utility in tissue engineering and the progress predicts a bright future. On this 30th anniversary of the Netherlands Biomaterials and Tissue Engineering society, we will briefly recount the state of supramolecular biomaterials in the Dutch academic and industrial research and development context. This review will provide the background, recent advances, industrial successes and challenges, as well as future directions of the field, as we see it. Throughout this work, we notice the intricate interplay between simplicity and complexity in creating more advanced solutions. We hope that the interplay and juxtaposition between these two forces can propel the field forward.

Protein biochips are at the heart of many medical and bioanalytical applications. Increasing interest has been focused on surface activation and subsequent functionalization strategies for immobilizing these biomolecules. Different approaches using covalent and noncovalent chemistry are reviewed; particular emphasis is placed on the chemical specificity of protein attachment and on retention of protein function. Strategies for creating protein patterns as opposed to protein arrays are also outlined. An outlook on promising and challenging future directions for protein biochip research and applications is also offered.

Proteinbiochips sind integrale Bestandteile einer wachsenden Zahl medizinischer und bioanalytischer Anwendungen, weshalb das Interesse an Methoden zur Proteinimmobilisierung für die Herstellung solcher Chips in den letzten Jahren stark gestiegen ist. Dieser Aufsatz gibt einen Überblick über chemische Verfahren zur kovalenten und nichtkovalenten Anbindung von Proteinen auf Oberflächen, wobei ein spezielles Augenmerk auf chemische Selektivität und die Erhaltung der Proteinfunktion bei der Immobilisierung gelegt wird. Ferner werden Strategien zur Herstellung strukturierter Proteinoberflächen umrissen. Abschließend wird ein Ausblick auf mögliche Entwicklungsrichtungen im Bereich der Proteinbiochipforschung und anwendung gegeben.

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With scanning tunneling microscopy (STM), the intramolecular conformational and intermolecular ordering aspects have been investigated of a variety of organic molecules physisorbed at the liquid-solid interface. By balancing the interplay between intramolecular and intermolecular interactions (hydrogen bonding), leading to control of the molecular conformation, foldamers were created which order into well-defined two-dimensional crystals. The nature of the hydrogen bonding groups in conjugated oligomers leads to the formation of infinite stacks and cyclic multimers, expressing the chiral nature of the molecules.