Red-light-controlled supramolecular assemblies of indigo amphiphiles at multiple length scales.

Amphiphilic molecules functionalized with photoresponsive motifs have attractive prospects for applications in smart functional bio-material ranging from cell-material interfaces to drug delivery systems owing to the precisely controllable functionality of self-assembled hierarchical supramolecular structures in aqueous media by a non-invasive light stimulation with high temporal- and spatial-resolution. However, most of reported photoresponsive amphiphiles are triggered by bio-damaging UV-light, which greatly limits the potential in bio-related applications. Herein, we present newly designed red-light controlled N,N'-diaryl-substituted indigo amphiphiles (IA), exhibiting excellent photoswitchablity and photostability with dual red-/green-light in organic media. Meanwhile, aqueous solutions of IA assembled into supramolecular structures in both microscopic and macroscopic length-scale, though the photoresponsiveness of IA is slightly compromised in aqueous media. At macroscopic length-scale, morphological changes of IA macroscopic scaffold prepared by a shear-flow method can be fine adjusted upon red-light irradiation. Moreover, the preferential attachment of live h-MSCs to IA macroscopic scaffold surface also indicates a good biocompatibility of IA macroscopic scaffold. These results provide the potential for developing the next generation of red-light controlled soft functional materials with good biocompatibility.

Tailoring Multicontrolled Supramolecular Assemblies of Stiff-Stilbene Amphiphiles into Macroscopic Soft Scaffolds as Cell-Material Interfaces.

Biocompatible synthetic supramolecular systems have shed light on biomedical and tissue-regenerative material applications. The intrinsic functional applicability, tunability, and stimuli-responsiveness of synthetic supramolecular systems allow one to develop various multicontrolled supramolecular assemblies in aqueous media. However, it remains highly challenging to use state-of-the-art supramolecular assemblies of photoresponsive amphiphiles controlled by multiple stimulations in fabricating macroscopic materials. Herein, we demonstrate a stiff-stilbene amphiphile (SA) multicontrolled supramolecular assembling system that comprises two different charged end groups. The excellent photoswitchabilities of SA in both organic and aqueous media are demonstrated. Furthermore, multiple stimuli, i.e., light, pH, and counterions, are applied to control the supramolecular assembling behaviors, which are monitored by circular dichroism spectroscopy and electron microscopies. This multicontrolled supramolecular system can be systematically assembled into macroscopic soft functional scaffolds, whose structural parameters are investigated by electron microscopies and X-ray diffraction techniques, suggesting the large aspect ratio of SA nanostructures assembled into macroscopic soft scaffolds. The fabricated soft functional scaffold is highly biocompatible for photocontrolled biotarget encapsulation/release selectively, as well as a cell-material interface for diverse cells' attachment. This new synthetic multicontrolled soft functional material provides a new strategy toward the development of next-generation controllable and biocompatible soft functional materials.

Synthesis and Characterization of Side-Chain Liquid-Crystalline Block Copolymers Containing Cyano-Terminated Phenyl Benzoate Moieties.

Block copolymers, known for their capacity to undergo microphase separation, spontaneously yield various periodic nanostructures. These precisely controlled nanostructures have attracted considerable interest due to their potential applications in microfabrication templates, conducting films, filter membranes, and other areas. However, it is crucial to acknowledge that microphase-separated structures typically exhibit random alignment, making alignment control a pivotal factor in functional material development. To address this challenge, researchers have explored the use of block copolymers containing liquid-crystalline (LC) polymers, which offer a promising technique for alignment control. The molecular structure and LC behavior of these polymers significantly impact the morphology and alignment of microphase-separated structures. In this study, we synthesized LC diblock copolymers with cyano-terminated phenyl benzoate moieties and evaluated the microphase-separated structures and molecular alignment behaviors. The LC diblock copolymers with a narrow molecular weight distribution were synthesized by atom transfer radical polymerization. Small angle X-ray scattering measurements revealed that the block copolymers exhibit smectic LC phases and form cylinder structures with a lattice period of about 18 nm by microphase separation. The examination of block copolymer films using polarized optical microscopy and polarized UV-visible absorption spectroscopy corroborated that the LC moieties were uniaxially aligned along the alignment treatment direction.

A Coformer Approach for Supramolecular Polymerization at High Concentrations.

Insolubility of functional molecules caused by polymorphism sometimes poses limitations for their solution-based processing. Such a situation can also occur in the preparation processes of supramolecular polymers formed in a solution. An effective strategy to address this issue is to prepare amorphous solid states by introducing a "coformer" molecule capable of inhibiting the formation of an insoluble polymorph through co-aggregation. Herein, inspired by the coformer approach, we demonstrated a solubility enhancement of a barbiturate π-conjugated compound that can supramolecularly polymerize through six-membered hydrogen-bonded rosettes. Our newly synthesized supramolecular coformer molecule features a sterically demanding methyl group in the π-conjugated unit of the parent molecule. Although the parent molecule exhibits low solubility in nonpolar solvents due to the formation of a crystalline polymorph comprising a tape-like hydrogen-bonded array prior to the supramolecular polymerization, mixing with the coformer compound enhanced the solubility by inhibiting mesoscopic organization of the tapes. The two monomers were then co-polymerized into desired helicoidal supramolecular polymers through the formation of heteromeric rosettes.

Collective bending motion of a two-dimensionally correlated bowl-stacked columnar liquid crystalline assembly under a shear force.

Stacked teacups inspired the idea that columnar assemblies of stacked bowl-shaped molecules may exhibit a unique dynamic behavior, unlike usual assemblies of planar disc- and rod-shaped molecules. On the basis of the molecular design concept for creating higher-order discotic liquid crystals, found in our group, we synthesized a sumanene derivative with octyloxycarbonyl side chains. This molecule forms an ordered hexagonal columnar mesophase, but unexpectedly, the columnar assembly is very soft, similar to sugar syrup. It displays, upon application of a shear force on solid substrates, a flexible bending motion with continuous angle variations of bowl-stacked columns while preserving the two-dimensional hexagonal order. In general, alignment control of higher-order liquid crystals is difficult to achieve due to their high viscosity. The present system that brings together higher structural order and mechanical softness will spark interest in bowl-shaped molecules as a component for developing higher-order liquid crystals with unique mechanical and stimuli-responsive properties.

Overcoming the entropy of polymer chains by making a plane with terminal groups: a thermoplastic PDMS with a long-range 1D structural order.

Due to its unique physical and chemical properties, polydimethylsiloxane (PDMS) is widely used in many applications, in which covalent cross-linking is commonly used to cure the fluidic polymer. The formation of a non-covalent network achieved through the incorporation of terminal groups that exhibit strong intermolecular interactions has also been reported to improve the mechanical properties of PDMS. Through the design of a terminal group capable of two-dimensional (2D) assembly, rather than the generally used multiple hydrogen bonding motifs, we have recently demonstrated an approach for inducing long-range structural ordering of PDMS, resulting in a dramatic change in the polymer from a fluid to a viscous solid. Here we present an even more surprising terminal-group effect: simply replacing a hydrogen with a methoxy group leads to extraordinary enhancement of the mechanical properties, giving rise to a thermoplastic PDMS material without covalent cross-linking. This finding would update the general notion that less polar and smaller terminal groups barely affect polymer properties. Based on a detailed study of the thermal, structural, morphological and rheological properties of the terminal-functionalized PDMS, we revealed that 2D assembly of the terminal groups results in networks of PDMS chains, which are arranged as domains with long-range one-dimensional (1D) periodic order, thereby increasing the storage modulus of the PDMS to exceed its loss modulus. Upon heating, the 1D periodic order is lost at around 120 °C, while the 2D assembly is maintained up to ∼160 °C. The 2D and 1D structures are recovered in sequence upon cooling. Due to the thermally reversible, stepwise structural disruption/formation as well as the lack of covalent cross-linking, the terminal-functionalized PDMS shows thermoplastic behavior and self-healing properties. The terminal group presented herein, which can form a 'plane', might also drive other polymers to assemble into a periodically ordered network structure, thereby allowing for significant modulation of their mechanical properties.

Individually separated supramolecular polymer chains toward solution-processable supramolecular polymeric materials.

Herein, we present a simple design concept for a monomer that affords individually separated supramolecular polymer chains. Random introduction of alkyl chains with different lengths onto a monomer prevented its supramolecular polymers from bundling, permitting the preparation of concentrated solutions of the supramolecular polymer without gelation, precipitation, or crystallization. With such a solution in hand, we succeeded in fabricating self-standing films and threads consisting of supramolecular polymers.

Amplification of Molecular Asymmetry during the Hierarchical Self-Assembly of Foldable Azobenzene Dyads into Nanotoroids and Nanotubes.

The amplification of molecular asymmetry through self-assembly is a phenomenon that not only comprehends the origin of homochirality in nature but also produces chiroptically active functional materials from molecules with minimal enantiomeric purity. Understanding how molecular asymmetry can be transferred and amplified into higher-order structures in a hierarchical self-assembly system is important but still unexplored. Herein, we present an intriguing example of the amplification of molecular asymmetry in hierarchically self-assembled nanotubes that feature discrete and isolatable toroidal intermediates. The hierarchical self-assembly is initiated via asymmetric intramolecular folding of scissor-shaped azobenzene dyads furnished with chiral side chains. When scalemic mixtures of the enantiomers are dissolved in a non-polar solvent and cooled to 20 °C, single-handed nanotoroids are formed, as confirmed using atomic force microscopy and circular dichroism analyses. A strong majority-rules effect at the nanotoroid level is observed and can be explained by a low mismatch penalty and a high helix-reversal penalty. The single-handed nanotoroids stack upon cooling to 0 °C to exclusively afford their respective single-handed nanotubes. Thus, the same degree of amplification of molecular asymmetry is realized at the nanotube level. The internal packing efficiency of molecules within nanotubes prepared from the pure enantiomers or their scalemic mixtures is likely different, as suggested by the absence of higher-order structure (supercoil) formation in the latter. X-ray diffraction analysis of the anisotropically oriented nanotube films revealed looser molecular packing within the scalemic nanotubes, which clearly reflects the lower enantiomeric purity of their internal chiral side chains.

Design of discotic liquid crystal enabling complete switching along with memory of homeotropic and homogeneous alignment over a large area.

The alignment control of discotic columnar liquid crystals (LCs), featuring a low motility of the constituent molecules and thus having a large viscosity, is a challenging task. Here we show that triphenylene hexacarboxylic ester, when functionalized with hybrid side chains consisting of alkyl and perfluoroalkyl groups in an appropriate ratio, gives a hexagonal columnar (Colh) LC capable of selectively forming large-area uniform homeotropic or homogeneous alignments, upon cooling from its isotropic melt or upon application of a shear force at its LC temperature, respectively. In addition to the alignment switching ability, each alignment state remains persistent unless the LC is heated to its melting temperature. In situ X-ray diffraction analysis under the application of a shear force, together with polarized optical microscopy observations, revealed how the columnar assembly is changed during the alignment-switching process. The remarkable behavior of the discotic LC is discussed in terms of its rheological properties.

Visible-light controlled supramolecular transformations of donor-acceptor Stenhouse adducts amphiphiles at multiple length-scale.

Designing responsive, adaptive, and dynamic supramolecular systems in water, the incorporation of photoresponsive units in amphiphilic molecular structures enables functional responses in a non-invasive way by using light. However, in aqueous media, vast majority of reported synthetic photoresponsive molecular amphiphiles are commonly driven by high energy and bio-damaging UV-light for supramolecular transformation at multiple length-scale. Herein, we present newly designed visible-light controlled supramolecular assembly of donor-acceptor Stenhouse adducts amphiphiles (DA) with excellent stability and solubility in aqueous media. The excellent photoswitchability in organic media and photoresponsiveness in aqueous media driven by visible-light are found, as confirmed with UV-vis absorption and NMR spectroscopies. Supramolecular assembly at multiple length-scale of DAs is investigated with electron microscopies and X-ray diffraction to show large aspect-ratio of nanostructures assembled into macroscopic soft scaffolds. Upon visible-light irradiation, the large geometrical transformation of DAs enables supramolecular transformations, and subsequently destabilizes the macroscopic soft scaffold to release fluorophores from the scaffolds. These results provide the feasibility in developing the next generation of visible-light controlled macroscopic soft functional scaffold from supramolecular assembly across multiple length-scale without and offer ample opportunity to design future soft robotic materials and functional biomaterials.

Supramolecular polymerization of thiobarbituric acid naphthalene dye.

2-Thiobarbituric acid-functionalized naphthalene dye selectively self-assembles into crystalline fibers to show material properties that are different from those of a previously reported oxo-barbituric acid derivative affording curved supramolecular polymers via the formation of hydrogen-bonded rosettes. Detailed studies using 1H-NMR, DFT calculation and X-ray diffraction revealed that the crystalline fiber is formed through a tape-like hydrogen-bonding pattern due to the weaker hydrogen-bond-acceptor ability of the thiocarbonyl group.

Aqueous Supramolecular Co-Assembly of Anionic and Cationic Photoresponsive Stiff-Stilbene Amphiphiles.

Fabrication of macroscopic soft functional materials, such as macroscopic photoresponsive soft materials and artificial muscles, can be commonly prepared by charge screening of supramolecular assemblies with inorganic salt solutions using a shear-flow method. However, some of the charged end-groups of photoresponsive molecular amphiphiles cannot be stabilized with inorganic salt solutions to fabricate macroscopic soft materials. Stiff stilbene amphiphiles (SAs) functionalized with anionic phosphite and cationic quaternary ammonium end groups are designed and synthesized and their photochemical and supramolecular assembly properties are determined. Supramolecular co-assembly of anionic and cationic nanotubes of SAs allows to transform into nanoribbons, confirmed by transmission electron microscopy, critical aggregation concentration, and Zeta potential measurements. Nanoribbons of anionic and cationic SAs can be prepared into macroscopic soft materials with inorganic salt solutions and surprisingly also with deionized water. The macroscopic soft material of anionic and cationic SAs can be stabilized at low concentration ≈5 mm. Meanwhile, the photoresponsiveness of the macroscopic soft materials is retained to provide macroscopic morphological change upon photoirradiation. These results exhibit the feasibility in fabrication of macroscopic functional soft materials from supramolecular assembly across multiple length-scale without help of inorganic salts and offer ample opportunity in developing future soft supramolecular robotic systems.

Sorafenib targets and inhibits the oncogenic properties of endometrial cancer stem cells via the RAF/ERK pathway.

BACKGROUND: Distinct subsets of cancer stem cells (CSCs) drive the initiation and progression of malignant tumors via enhanced self-renewal and development of treatment/apoptosis resistance. Endometrial CSC-selective drugs have not been successfully developed because most endometrial cell lines do not contain a sufficient proportion of stable CSCs. Here, we aimed to identify endometrial CSC-containing cell lines and to search for endometrial CSC-selective drugs. METHODS: We first assessed the presence of CSCs by identifying side populations (SPs) in several endometrial cancer cell lines. We then characterized cell viability, colony-formation, transwell invasion and xenotransplantion capability using the isolated SP cells. We also conducted real-time RT-PCR, immunoblot and immunofluorescence analyses of the cells' expression of CSC-associated markers. Focusing on 14 putative CSC-selective drugs, we characterized their effects on the proliferation and apoptosis of endometrial cancer cell lines, examining cell viability and annexin V staining. We further examined the inhibitory effects of the selected drugs, focusing on proliferation, invasion, expression of CSC-associated markers and tumor formation. RESULTS: We focused on HHUA cells, an endometrial cancer cell line derived from a well-differentiated endometrial adenocarcinoma. HHUA cells contained a sufficient proportion of stable CSCs with an SP phenotype (HHUA-SP). HHUA-SP showed greater proliferation, colony-formation, and invasive capabilities compared with the main population of HHUA cells (HHUA-MP). HHUA-SP generated larger tumors with higher expression of proliferation-related markers, Ki67, c-MYC and phosphorylated ERK compared with HHUA-MP when transplanted into immunodeficient mice. Among the 14 candidate drugs, sorafenib, an inhibitor of RAF pathways and multiple kinase receptors, inhibited cell proliferation and invasion in both HHUA-SP and -MP, but more profoundly in HHUA-SP. In vivo treatment with sorafenib for 4 weeks reduced the weights of HHUA-SP-derived tumors and decreased the expression of Ki67, ZEB1, and RAF1. CONCLUSIONS: Our results suggest that HHUA is a useful cell line for discovery and identification of endometrial CSC-selective drugs, and that sorafenib may be an effective anti-endometrial cancer drug targeting endometrial CSCs.

Photoactuating Artificial Muscles of Motor Amphiphiles as an Extracellular Matrix Mimetic Scaffold for Mesenchymal Stem Cells.

Mimicking the native extracellular matrix (ECM) as a cell culture scaffold has long attracted scientists from the perspective of supramolecular chemistry for potential application in regenerative medicine. However, the development of the next-generation synthetic materials that mimic key aspects of ECM, with hierarchically oriented supramolecular structures, which are simultaneously highly dynamic and responsive to external stimuli, remains a major challenge. Herein, we present supramolecular assemblies formed by motor amphiphiles (MAs), which mimic the structural features of the hydrogel nature of the ECM and additionally show intrinsic dynamic behavior that allow amplifying molecular motions to macroscopic muscle-like actuating functions induced by light. The supramolecular assembly (named artificial muscle) provides an attractive approach for developing responsive ECM mimetic scaffolds for human bone marrow-derived mesenchymal stem cells (hBM-MSCs). Detailed investigations on the photoisomerization by nuclear magnetic resonance and UV-vis absorption spectroscopy, assembled structures by electron microscopy, the photoactuation process, structural order by X-ray diffraction, and cytotoxicity are presented. Artificial muscles of MAs provide fast photoactuation in water based on the hierarchically anisotropic supramolecular structures and show no cytotoxicity. Particularly important, artificial muscles of MAs with adhered hBM-MSCs still can be actuated by external light stimulation, showing their ability to convert light energy into mechanical signals in biocompatible systems. As a proof-of-concept demonstration, these results provide the potential for building photoactuating ECM mimetic scaffolds by artificial muscle-like supramolecular assemblies based on MAs and offer opportunities for signal transduction in future biohybrid systems of cells and MAs.

Changing the structural and physical properties of 3-arm star poly(δ-valerolactone)s by a branch-point design.

The chemical structure of a branch point of star-shaped polymers has been considered to have a small influence on the physical properties of the entire polymer. Contrary to this general notion, here we show that a 3-arm star polymer, composed of three poly(δ-valerolactone) arms extended from one side of a triptycene branch point, exhibits a remarkably high complex viscosity, compared to the analogous star-shaped polymers with a branch point of a triptycene isomer or triphenylethane.

Introduction of Triptycene with a Particular Substitution Pattern into Polymer Chains Can Dramatically Improve the Structural and Rheological Properties.

Although a large number of polymers that contain triptycene units in the main chains have been developed, no polymer design using 1,8-substituted triptycene has been reported to date. In this study, we investigated the properties of linear homo- and copolymers obtained by ring-opening polymerization of a triptycene monomer bearing a macrocyclic olefin linked at its 1,8-position and its copolymerization with cyclooctene, respectively. We found that the introduction of triptycene with this substitution pattern leads to nanoscale molecular ordering, thereby greatly improving the physical properties of the polymers. The key to this remarkable behavior of 1,8-substituted triptycene-containing polymers is the formation of a particular two-dimensional assembly of the triptycene units by nested hexagonal packing, which aligns one-dimensionally while folding the polymer chains into a well-defined layered structure. The polymer design using 1,8-substituted triptycene can be applied to other polymers, unless their main chain contains functional groups capable of a strong intermolecular interaction such as hydrogen bonding.

Room-Temperature Pentacene Fluids: Oligoethylene Glycol Substituent-Controlled Morphologies and Singlet Fission.

We report the first synthesis of solvent-free pentacene fluids at room temperature together with observation of singlet fission (SF). Three pentacenes with different number of ethylene glycol (EG) side chains (n) were employed (denoted as (EG)n-Pc-(EG)n: n = 2, 3, and 4). The morphologies of these pentacenes largely depend on the lengths of EG chains (n). (EG)3-Pc-(EG)3 and (EG)4-Pc-(EG)4 indicate fluid compounds at room temperature, whereas (EG)2-Pc-(EG)2 is a solid compound. Microscopic clustering with short-range interactions between pentacene chromophores was confirmed in X-ray diffraction profiles of solvent-free fluids. Such a structural trend is an important origin of SF and consistent with the steady-state spectroscopic results. To one's surprise, femtosecond transient absorption spectroscopy demonstrated that SF occurred in thin films prepared from solvent-free fluids of (EG)3-Pc-(EG)3 and (EG)4-Pc-(EG)4 in spite of such excessive EG chains.

Self-assembly of alkylated and perfluoroalkylated scissor-shaped azobenzene dyads into distinct structures.

Scissor-shaped azobenzene dyads possessing alkyl side chains can hierarchically self-assemble through a folded conformation into toroidal and tubular nanostructures. We found that the introduction of perfluoroalkyl side chains transforms the assembly pathway into direct one-dimensional stacking of the folded conformer, resulting in the formation of gel-forming supramolecular fibers that can reversibly dissociate and reassemble on exposure to light.

Hydrogen bond-directed supramolecular polymorphism leading to soft and hard molecular ordering.

Transformation of metastable supramolecular stacks of hydrogen-bonded rosettes composed of an ester-containing barbiturated naphthalene into crystalline nanosheets occurs through the rearrangement of hydrogen-bonding patterns. The involvement of the ester group in the crystalline hydrogen-bonded pattern is demonstrated, guiding us to a new molecular design that can afford supramolecular polymorphs with soft and hard molecular packing.

Design of a molecular memory element with an alternating circular array of dipolar rotors and rotation suppressors.

As a new element for electric-field driven molecular memory, we developed a hexaarylbenzene derivative in which three difluorophenyl groups and three aryl groups as a dipolar rotor and a rotation suppressor, respectively, are alternately positioned on the central benzene core. This molecule has two rotational isomeric forms, both of which preserve their conformational states at room temperature but exhibit interconversion at high temperatures. Amorphous thin films fabricated from the hexaarylbenzene show a reversible change in surface potential by application of electric fields.