Mastering Dendrimer Self‐Assembly for Efficient siRNA Delivery: From Conceptual Design to In Vivo Efficient Gene Silencing

Self‐assembly is a fundamental concept and a powerful approach in molecular science. However, creating functional materials with the desired properties through self‐assembly remains challenging. In this work, through a combination of experimental and computational approaches, the self‐assembly of small amphiphilic dendrons into nanosized supramolecular dendrimer micelles with a degree of structural definition similar to traditional covalent high‐generation dendrimers is reported. It is demonstrated that, with the optimal balance of hydrophobicity and hydrophilicity, one of the self‐assembled nanomicellar systems, totally devoid of toxic side effects, is able to deliver small interfering RNA and achieve effective gene silencing both in cells – including the highly refractory human hematopoietic CD34⁺ stem cells – and in vivo, thus paving the way for future biomedical implementation. This work presents a case study of the concept of generating functional supramolecular dendrimers via self‐assembly. The ability of carefully designed and gauged building blocks to assemble into supramolecular structures opens new perspectives on the design of self‐assembling nanosystems for complex and functional applications.     

Pick‐and‐Place Assembly of Single Microtubules

Intracellular transport is affected by the filament network in the densely packed cytoplasm. Biophysical studies focusing on intracellular transport based on microtubule–kinesin system frequently use in vitro motility assays, which are performed either on individual microtubules or on random (or simple) microtubule networks. Assembling intricate networks with high flexibility requires the manipulation of 25 nm diameter microtubules individually, which can be achieved through the use of pick‐and‐place assembly. Although widely used to assemble tiny objects, pick‐and‐place is not a common practice for the manipulation of biological materials. Using the high‐level handling capabilities of microelectromechanical systems (MEMS) technology, tweezers are designed and fabricated to pick and place single microtubule filaments. Repeated picking and placing cycles provide a multilayered and multidirectional microtubule network even for different surface topographies. On‐demand assembly of microtubules forms crossings at desired angles for biophysical studies as well as complex networks that can be used as nanotransport systems.     

Precise Self‐Assembly of Nanoparticles into Ordered Nanoarchitectures Directed by Tobacco Mosaic Virus Coat Protein

Self‐assembly guided by biological molecules is a promising approach for fabricating predesigned nanostructures. Protein is one such biomolecule possessing deterministic 3D crystal structure and peptide information, which acts as a good candidate for templating functional nanoparticles (fNPs). However, inadequate coordination efficacy during the establishment of interfacial interactions with fNPs makes it highly challenging to precisely fabricate designed nanostructures and functional materials. Here, a facile and robust strategy is reported for the hierarchical assembly of fNPs into ordered architectures, with unprecedentedly large sizes up to tens of micrometers, using a hollow cylinder‐shaped tobacco mosaic virus coat protein (TMV disk). The rational design of the site‐specific functional groups on the TMV disk not only demonstrates the powerful capability of directing various discrete fNP assemblies with high controllability but also assists in precise assembly of a TMV monolayer sheet structure for further organizing homogeneous and heterogeneous fNP periodic lattices by varying the types of fNPs. The high precision and adjustability of the pattern fashions of different fNPs unambiguously corroborate the validity of this innovative strategy, which provides a convenient route to design and assemble protein‐based hierarchical ordered architectures for use in nanophotonics and nanodevices.     

Encapsulation of Gold Nanoparticles into DNA Minimal Cages for 3D‐Anisotropic Functionalization and Assembly

Gold nanoparticles (AuNPs) endowed with anisotropic DNA valency are an important class of materials, as they can assemble into complex structures with a minimal number of DNA strands. However, methods to encode 3D DNA strand patterns on AuNPs with a controlled number of unique DNA strands in a predesigned spatial arrangement remain elusive. In this work, a simple one‐step method to yield such DNA‐decorated AuNPs is demonstrated, through encapsulating AuNPs into DNA minimal nanocages. The AuNP@DNA cage encapsulation complex inherits the 3D anisotropic molecular information from the DNA nanocage with enhanced structural stability. The DNA nanocage can be further functionalized and used as a building block for the self‐assembly of complex architectures, such as dimers and trimers, programmed assemblies with sequential growth DNA backbones and DNA origami.     

Independent Blue and Red Light Triggered Narcissistic Self‐Sorting Self‐Assembly of Colloidal Particles

The ability of living systems to self‐sort different cells into separate assemblies and the ability to independently regulate different structures are one ingredient that gives rise to their spatiotemporal complexity. Here, this self‐sorting behavior is replicated in a synthetic system with two types of colloidal particles; where each particle type independently self‐assembles either under blue or red light into distinct clusters, known as narcissistic self‐sorting. For this purpose, each particle type is functionalized either with the light‐switchable protein VVDHigh or Cph1, which homodimerize under blue and red light, respectively. The response to different wavelengths of light and the high specificity of the protein interactions allows for the independent self‐assembly of each particle type with blue or red light and narcissistic self‐sorting. Moreover, as both of the photoswitchable protein interactions are reversible in the dark; also, the self‐sorting is reversible and dynamic. Overall, the independent blue and red light controlled self‐sorting in a synthetic system opens new possibilities to assemble adaptable, smart, and advanced materials similar to the complexity observed in tissues.     

Scaffold‐Free Liver‐On‐A‐Chip with Multiscale Organotypic Cultures

The considerable advances that have been made in the development of organotypic cultures have failed to overcome the challenges of expressing tissue‐specific functions and complexities, especially for organs that require multitasking and complex biological processes, such as the liver. Primary liver cells are ideal biological building blocks for functional organotypic reconstruction, but are limited by their rapid loss of physiological integrity in vitro. Here the concept of lattice growth used in material science is applied to develop a tissue incubator, which provides physiological cues and controls the 3D assembly of primary cells. The cues include a biological growing template, spatial coculture, biomimetic radial flow, and circulation in a scaffold‐free condition. The feasibility of recapitulating a multiscale physiological structural hierarchy, complex drug clearance, and zonal physiology from the cell to tissue level in long‐term cultured liver‐on‐a‐chip is demonstrated. These methods are promising for future applications in pharmacodynamics and personal medicine.     

Programmable Construction of Peptide‐Based Materials in Living Subjects: From Modular Design and Morphological Control to Theranostics

Self‐assembled nanomaterials show potential high efficiency as theranostics for high‐performance bioimaging and disease treatment. However, the superstructures of pre‐assembled nanomaterials may change in the complicated physiological conditions, resulting in compromised properties and/or biofunctions. Taking advantage of chemical self‐assembly and biomedicine, a new strategy of “in vivo self‐assembly” is proposed to in situ construct functional nanomaterials in living subjects to explore new biological effects. Herein, recent advances on peptide‐based nanomaterials constructed by the in vivo self‐assembly strategy are summarized. Modular peptide building blocks with various functions, such as targeting, self‐assembly, tailoring, and biofunctional motifs, are employed for the construction of nanomaterials. Then, self‐assembly of these building blocks in living systems to construct various morphologies of nanostructures and corresponding unique biological effects, such as assembly/aggregation‐induced retention (AIR), are introduced, followed by their applications in high‐performance drug delivery and bioimaging. Finally, an outlook and perspective toward future developments of in vivo self‐assembled peptide‐based nanomaterials for translational medicine are concluded.     

Bottom‐Up Assembly of Functional Intracellular Synthetic Organelles by Droplet‐Based Microfluidics

Bottom‐up synthetic biology has directed most efforts toward the construction of artificial compartmentalized systems that recreate living cell functions in their mechanical, morphological, or metabolic characteristics. However, bottom‐up synthetic biology also offers great potential to study subcellular structures like organelles. Because of their intricate and complex structure, these key elements of eukaryotic life forms remain poorly understood. Here, the controlled assembly of lipid enclosed, organelle‐like architectures is explored by droplet‐based microfluidics. Three types of giant unilamellar vesicles (GUVs)‐based synthetic organelles (SOs) functioning within natural living cells are procedured: (A) synthetic peroxisomes supporting cellular stress‐management, mimicking an organelle innate to the host cell by using analogous enzymatic modules; (B) synthetic endoplasmic reticulum (ER) as intracellular light‐responsive calcium stores involved in intercellular calcium signalling, mimicking an organelle innate to the host cell but utilizing a fundamentally different mechanism; and (C) synthetic magnetosomes providing eukaryotic cells with a magnetotactic sense, mimicking an organelle that is not natural to the host cell but transplanting its functionality from other branches of the phylogenetic tree. Microfluidic assembly of functional SOs paves the way for high‐throughput generation of versatile intracellular structures implantable into living cells. This in‐droplet SO design may support or expand cellular functionalities in translational nanomedicine.     

Photo‐Induced Polymerization and Reconfigurable Assembly of Multifunctional Ferrocene‐Tyrosine

The photo‐induced reconfigurable assembly of nanostructures via the simultaneous noncovalent and covalent polymerization of a functional ferrocene‐tyrosine (Fc‐Y) molecule is reported. The Fc‐Y monomers can directly self‐assemble into nanospheres with a smooth surface driven by noncovalent interactions. By covalent photo‐crosslinking of the Fc‐Y monomers, the nanospheres transform spontaneously into hollow vesicles composed of hierarchically ordered lamellar structures. It is worth noting that the formed nanostructures exhibit both reducing property for in situ mineralization of gold nanoparticles with tunable biocatalytic behavior, and the redox activity for superior energy storage capacity. The measured energy storage capacity is 31 mAh g⁻¹ for the nanospheres, which is the highest value reported so far for peptide assemblages as supercapacitor. The results offer insights into the dynamic self‐assembly of highly ordered multifunctional materials with promising applications in catalysis, sensing, energy and biomedical fields.     

Supramolecular Two-Dimensional Systems and Their Biological Applications

Various biological systems rely on the supramolecular assembly of biomolecules through noncovalent bonds for performing sophisticated functions. In particular, cell membranes, which are 2D structures in biological systems, have various characteristics such as a large surface, flexibility, and molecule-recognition ability. Supramolecular 2D materials based on biological systems provide a novel perspective for the development of functional 2D materials. The physical and chemical properties of 2D structures, attributed to their large surface area, can enhance the sensitivity of the detection of target molecules, molecular loading, and bioconjugation efficiency, suggesting the potential utility of functional 2D materials as candidates for biological systems. Although several types of studies on supramolecular 2D materials have been reported, supramolecular biofunctional 2D materials have not been reviewed previously. In this regard, the current advances in 2D material development using molecular assembly are discussed with respect to the rational design of self-assembling aromatic amphiphiles, the formation of 2D structures, and the biological applications of functional 2D materials.     

Under Diffusion Control: from Structuring Matter to Directional Motion

Self‐organization in synthetic chemical systems is quickly developing into a powerful strategy for designing new functional materials. As self‐organization requires the system to exist far from thermodynamic equilibrium, chemists have begun to go beyond the classical equilibrium self‐assembly that is often applied in bottom‐up supramolecular synthesis, and to learn about the surprising and unpredicted emergent properties of chemical systems that are characterized by a higher level of complexity and extended reactivity networks. The present review focuses on self‐organization in reaction‐diffusion systems. Selected examples show how the emergence of complex morphogenesis is feasible in synthetic systems leading to hierarchically and nanostructured matter. Starting from well‐investigated oscillating reactions, recent developments extend diffusion‐limited reactivity to supramolecular systems. The concept of dynamic instability is introduced and illustrated as an additional tool for the design of smart materials and actuators, with emphasis on the realization of motion even at the macroscopic scale. The formation of spatio‐temporal patterns along diffusive chemical gradients is exploited as the main channel to realize symmetry breaking and therefore anisotropic and directional mechanical transformations. Finally, the interaction between external perturbations and chemical gradients is explored to give mechanistic insights in the design of materials responsive to external stimuli.     

Biphasic Supramolecular Self‐Assembly of Ferric Ions and Tannic Acid across Interfaces for Nanofilm Formation

Cell nanoencapsulation provides a chemical tool for the isolation and protection of living cells from harmful, and often lethal, external environments. Although several strategies are available to form nanometric films, most methods heavily rely on time‐consuming multistep processes and are not biocompatible. Here, the interfacial supramolecular self‐assembly and film formation of ferric ions (Fe^III) and tannic acid (TA) in biphasic systems is reported, where Fe^III and TA come into contact each other and self‐assemble across the interface of two immiscible phases. The interfacial nanofilm formation developed is simple, fast, and cytocompatible. Its versatility is demonstrated with various biphasic systems: hollow microcapsules, encasing microbial or mammalian cells, that are generated at the water–oil interface in a microfluidic device; a cytoprotective Fe^III–TA shell that forms on the surface of the alginate microbead, which then entraps probiotic Lactobacillus acidophilus; and a pericellular Fe^III–TA shell that forms on individual Saccharomyces cerevisiae. This biphasic interfacial reaction system provides a simple but versatile structural motif in materials science, as well as advancing chemical manipulability of living cells.     

Printing Polymer Nanocomposites and Composites in Three Dimensions

Recent advances in materials science and three‐dimensional (3D) printing hold great promises to conceive new classes of multifunctional materials and components for functional devices and products. Various functionalities (e.g., mechanical, electrical, and thermal properties, magnetism) can be offered by the nano‐ and micro‐reinforcements to the non‐functional pure printing materials for the realization of advanced materials and innovative systems. In addition, the ability to print 3D structures in a layer‐by‐layer manner enables manufacturing of highly‐customized complex features and allows an efficient control over the properties of fabricated structures. Here, the authors present a brief overview mainly over the latest progresses in 3D printing of multifunctional polymer nanocomposites and microfiber‐reinforced composites including the benefits, limitations, and potential applications. Only those 3D printing techniques that are compatible with polymer nanocomposites and composites, that is, materials that have already been used as printing materials, are introduced. The very hot topic of 3D printing of thermoplastic composites featuring continuous microfibers is also briefly introduced.     

Beta‐Sheet‐Forming,Self‐Assembled Peptide Nanomaterials towards Optical,Energy, and Healthcare Applications

Peptide self‐assembly is an attractive route for the synthesis of intricate organic nanostructures that possess remarkable structural variety and biocompatibility. Recent studies on peptide‐based, self‐assembled materials have expanded beyond the construction of high‐order architectures; they are now reporting new functional materials that have application in the emerging fields such as artificial photosynthesis and rechargeable batteries. Nevertheless, there have been few reviews particularly concentrating on such versatile, emerging applications. Herein, recent advances in the synthesis of self‐assembled peptide nanomaterials (e.g., cross β‐sheet‐based amyloid nanostructures, peptide amphiphiles) are selectively reviewed and their new applications in diverse, interdisciplinary fields are described, ranging from optics and energy storage/conversion to healthcare. The applications of peptide‐based self‐assembled materials in unconventional fields are also highlighted, such as photoluminescent peptide nanostructures, artificial photosynthetic peptide nanomaterials, and lithium‐ion battery components. The relation of such functional materials to the rapidly progressing biomedical applications of peptide self‐assembly, which include biosensors/chips and regenerative medicine, are discussed. The combination of strategies shown in these applications would further promote the discovery of novel, functional, small materials.     

Human‐on‐Leaf‐Chip: A Biomimetic Vascular System Integrated with Chamber‐Specific Organs

The vascular network is a central component of the organ‐on‐a‐chip system to build a 3D physiological microenvironment with controlled physical and biochemical variables. Inspired by ubiquitous biological systems such as leaf venation and circulatory systems, a fabrication strategy is devised to develop a biomimetic vascular system integrated with freely designed chambers, which function as niches for chamber‐specific vascularized organs. As a proof of concept, a human‐on‐leaf‐chip system with biomimetic multiscale vasculature systems connecting the self‐assembled 3D vasculatures in chambers is fabricated, mimicking the in vivo complex architectures of the human cardiovascular system connecting vascularized organs. Besides, two types of vascularized organs are built independently within the two halves of the system to verify its feasibility for conducting comparative experiments for organ‐specific metastasis studies in a single chip. Successful culturing of human hepatoma G2 cells (HepG2s) and mesenchymal stem cells (MSCs) with human umbilical vein endothelial cells (HUVECs) shows good vasculature formation, and organ‐specific metastasis is simulated through perfusion of pancreatic cancer cells and shows distinct cancer encapsulation by MSCs, which is absent in HepG2s. Given good culture efficacy, study design flexibility, and ease of modification, these results show that the bioinspired human‐on‐leaf‐chip possesses great potential in comparative and metastasis studies while retaining organ‐to‐organ crosstalk.     

Magnetic Plasmon Networks Programmed by Molecular Self‐Assembly

Nanoscale manipulation of magnetic fields has been a long‐term pursuit in plasmonics and metamaterials, as it can enable a range of appealing optical properties, such as high‐sensitivity circular dichroism, directional scattering, and low‐refractive‐index materials. Inspired by the natural magnetism of aromatic molecules, the cyclic ring cluster of plasmonic nanoparticles (NPs) has been suggested as a promising architecture with induced unnatural magnetism, especially at visible frequencies. However, it remains challenging to assemble plasmonic NPs into complex networks exhibiting strong visible magnetism. Here, a DNA‐origami‐based strategy is introduced to realize molecular self‐assembly of NPs forming complex magnetic architectures, exhibiting emergent properties including anti‐ferromagnetism, purely magnetic‐based Fano resonances, and magnetic surface plasmon polaritons. The basic building block, a gold NP (AuNP) ring consisting of six AuNP seeds, is arranged on a DNA origami frame with nanometer precision. The subsequent hierarchical assembly of the AuNP rings leads to the formation of higher‐order networks of clusters and polymeric chains. Strong emergent plasmonic properties are induced by in situ growth of silver upon the AuNP seeds. This work may facilitate the development of a tunable and scalable DNA‐based strategy for the assembly of optical magnetic circuitry, as well as plasmonic metamaterials with high fidelity.     

Artificial Biosystems by Printing Biology

The continuous progress of printing technologies over the past 20 years has fueled the development of a plethora of applications in materials sciences, flexible electronics, and biotechnologies. More recently, printing methodologies have started up to explore the world of Artificial Biology, offering new paradigms in the direct assembly of Artificial Biosystems (small condensates, compartments, networks, tissues, and organs) by mimicking the result of the evolution of living systems and also by redesigning natural biological systems, taking inspiration from them. This recent progress is reported in terms of a new field here defined as Printing Biology, resulting from the intersection between the field of printing and the bottom up Synthetic Biology. Printing Biology explores new approaches for the reconfigurable assembly of designed life‐like or life‐inspired structures. This work presents this emerging field, highlighting its main features, i.e., printing methodologies (from 2D to 3D), molecular ink properties, deposition mechanisms, and finally the applications and future challenges. Printing Biology is expected to show a growing impact on the development of biotechnology and life‐inspired fabrication.     

Protein Bricks: 2D and 3D Bio‐Nanostructures with Shape and Function on Demand

Precise patterning of polymer‐based biomaterials for functional bio‐nanostructures has extensive applications including biosensing, tissue engineering, and regenerative medicine. Remarkable progress is made in both top‐down (based on lithographic methods) and bottom‐up (via self‐assembly) approaches with natural and synthetic biopolymers. However, most methods only yield 2D and pseudo‐3D structures with restricted geometries and functionalities. Here, it is reported that precise nanostructuring on genetically engineered spider silk by accurately directing ion and electron beam interactions with the protein's matrix at the nanoscale to create well‐defined 2D bionanopatterns and further assemble 3D bionanoarchitectures with shape and function on demand, termed “Protein Bricks.” The added control over protein sequence and molecular weight of recombinant spider silk via genetic engineering provides unprecedented lithographic resolution (approaching the molecular limit), sharpness, and biological functions compared to natural proteins. This approach provides a facile method for patterning and immobilizing functional molecules within nanoscopic, hierarchical protein structures, which sheds light on a wide range of biomedical applications such as structure‐enhanced fluorescence and biomimetic microenvironments for controlling cell fate.     

3D Lattice Engineering of Nanoparticles by DNA Shells

With the development of structural DNA nanotechnology, DNA has now far exceeded its original function: as a genetic code. It can, in principle, self‐assemble into desired shapes with accurate size. Moreover, it can perform as a functional linker to program other materials by grafting DNA onto these materials. Nanoparticles, both inorganic and organic, can now be programmatically assembled into complex 3D superlattices with high order when guided by DNA. By encoding functions into the as‐assembled nanoparticles, materials with excellent collective effects may be invented. Here, how nanoparticles with different shapes or functions are successfully fabricated into 3D lattices with the help of DNA shells coated on the surface and how scientists can produce desired lattices by design are reviewed. The cases to achieve dynamic superlattices of nanoparticles by affecting the environment where DNA survives are also discussed.     

Efficient Cisplatin Pro‐Drug Delivery Visualized with Sub‐100 nm Resolution: Interfacing Engineered Thermosensitive Magnetomicelles with a Living System

Temperature‐responsive magnetic nanomicelles can serve as thermal energy and cargo carriers with controlled drug release functionality. In view of their potential biomedical applications, understanding the modes of interaction between nanomaterials and living systems and evaluation of efficiency of cargo delivery is of the utmost importance. In this work, we investigate the interaction between the hybrid magnetic nanomicelles engineered for controlled platinum complex drug delivery and a biological system at three fundamental levels: subcellular compartments, a single cell and whole living animal. Nanomicelles with polymeric P(NIPAAm‐co‐AAm)‐b‐PCL core‐shell were loaded with a hydrophobic Pt(IV) complex and Fe₃O₄ nanoparticles though self‐assembly. The distribution of a platinum complex on subcellular level is visualized using hard X‐ray fluorescence microscopy with unprecedented level of detail at sub‐100 nm spatial resolution. We then study the cytotoxic effects of platinum complex‐loaded micelles in vitro on a head and neck cancer cell culture model SQ20B. Finally, by employing the magnetic functionality of the micelles and additionally loading them with a near infrared fluorescent dye, we magnetically target them to a tumor site in a live animal xenografted model which allows to visualize their biodistribution in vivo.