On‐chip technology for single‐cell arraying,electrorotation‐based analysis and selective release

This paper reports a method for label‐free single‐cell biophysical analysis of multiple cells trapped in suspension by electrokinetic forces. Tri‐dimensional pillar electrodes arranged along the width of a microfluidic chamber define actuators for single cell trapping and selective release by electrokinetic force. Moreover, a rotation can be induced on the cell in combination with a negative DEP force to retain the cell against the flow. The measurement of the rotation speed of the cell as a function of the electric field frequency define an electrorotation spectrum that allows to study the dielectric properties of the cell. The system presented here shows for the first time the simultaneous electrorotation analysis of multiple single cells in separate micro cages that can be selectively addressed to trap and/or release the cells. Chips with 39 micro‐actuators of different interelectrode distance were fabricated to study cells with different sizes. The extracted dielectric properties of Henrietta Lacks, human embryonic kidney 293, and human immortalized T lymphocytes cells were found in agreements with previous findings. Moreover, the membrane capacitance of M17 neuroblastoma cells was investigated and found to fall in in the range of 7.49 ± 0.39 mF/m².     

Efficient Release of Affinity‐Captured Cells Using a Coiled‐Coil‐Based Molecular Trigger

Affinity‐based cell separation is label‐free and highly specific, but it is difficult to efficiently and gently release affinity‐captured cells due to the multivalent nature of cell‐material interactions. To address this challenge, we have developed a platform composed of a capture substrate and a cell‐releasing molecular trigger. The capture substrate is functionalized with a cell‐capture antibody and a coiled‐coil A . The cell‐releasing molecular trigger B ‐PEG (polyethylene glycol), a conjugate of a coiled‐coil B and polyethylene glycol, can drive efficient and gentle release of the captured cells, because A / B heterodimerization brings B ‐PEG to the substrate and PEG chains adopt extended conformations and break nearby multivalent antibody‐biomarker interactions. No enzymes or excessive shear stress are involved, and the released cells have neither external molecules attached nor endogenous cell‐surface molecules cleaved, which is critical for the viability, phenotype, and function of sensitive cells.

     

Ile‐Lys‐Val‐ala‐Val (IKVAV) peptide for neuronal tissue engineering

Despite the great advances in microsurgery, some neural injuries cannot be treated surgically. Stem cell therapy is a potential approach for treating neuroinjuries and neurodegenerative disease. Researchers have developed various bioactive scaffolds for tissue engineering, exhibiting enhanced cell viability, attachment, migration, neurite elongation, and neuronal differentiation, with the aim of developing functional tissue grafts that can be incorporated in vivo. Facilitating the appropriate interactions between the cells and extracellular matrix is crucial in scaffold design. Modification of scaffolds with biofunctional motifs such as growth factors, drugs, or peptides can improve this interaction. In this review, we focus on the laminin‐derived Ile‐Lys‐Val‐Ala‐Val peptide as a biofunctional epitope for neuronal tissue engineering. Inclusion of this bioactive peptide within a scaffold is known to enhance cell adhesion as well as neuronal differentiation in both 2‐dimensional and 3‐dimensional environments. The in vivo application of this peptide is also briefly described.     

Coating Strategies Using Layer‐by‐layer Deposition for Cell Encapsulation

The layer‐by‐layer (LbL) deposition technique is widely used to develop multilayered films based on the directed assembly of complementary materials. In the last decade, thin multilayers prepared by LbL deposition have been applied in biological fields, namely, for cellular encapsulation, due to their versatile processing and tunable properties. Their use was suggested as an alternative approach to overcome the drawbacks of bulk hydrogels, for endocrine cells transplantation or tissue engineering approaches, as effective cytoprotective agents, or as a way to control cell division. Nanostructured multilayered materials are currently used in the nanomodification of the surfaces of single cells and cell aggregates, and are also suitable as coatings for cell‐laden hydrogels or other biomaterials, which may later be transformed to highly permeable hollow capsules. In this Focus Review, we discuss the applications of LbL cell encapsulation in distinct fields, including cell therapy, regenerative medicine, and biotechnological applications. Insights regarding practical aspects required to employ LbL for cell encapsulation are also provided.     

Synthesis and properties of mono‐ and di‐fluoro‐substituted 2,3‐didodecylquinoxaline‐based polymers for polymer solar cells

We synthesized two new alternating polymers, namely P(Tt‐FQx) and P(Tt‐DFQx) , incorporating electron rich tri‐thiophene and electron deficient 6‐fluoroquinoxaline or 6,7‐difluoroquinoxaline derivatives. Both polymers P(Tt‐FQx) and P(Tt‐DFQx) exhibited high thermal stabilities and the estimated 5% weight loss temperatures are 425 and 460 °C, respectively. Polymers P(Tt‐FQx) and P(Tt‐DFQx) displayed intense absorption band between 450 and 700 nm with an optical band gap (E_g) of 1.78 and 1.80 eV, respectively. The determined highest occupied/lowest unoccupied molecular orbital's (HOMO/LUMO) of P(Tt‐DFQx) (?5.48 eV/?3.68 eV) are slightly deeper than those of P(Tt‐FQx) ( ?5.32 eV/?3.54 eV). The polymer solar cells fabricated with a device structure of ITO/PEDOT:PSS/ P(Tt‐FQx) or P(Tt‐DFQx) :PC₇₀BM (1:1.5 wt %) + 3 vol % DIO/Al offered a maximum power conversion efficiency (PCE) of 3.65% with an open‐circuit voltage (V_oc) of 0.59 V, a short‐circuit current (J_sc) of 10.65 mA/cm² and fill factor (FF) of 59% for P(Tt‐FQx) ‐based device and a PCE of 4.36% with an V_oc of 0.69 V, a J_sc of 9.92 mA/cm², and FF of 63% for P(Tt‐DFQx) ‐based device. © 2018 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2019 , 57, 545–552     

Folic‐Acid‐Modified Conducting Polymer: Electrochemical Detection of the Cell Attachment

Here, postfunctionalization and bioapplication of a π‐conjugated polymer named 4‐[4H‐dithieno(3,2‐b:2′,3′‐d)pyrrol‐4‐yl]aniline (DTP‐aryl‐NH₂) are reported, which is successfully synthesized via electropolymerization onto the glassy carbon electrode. Folic acid (FA) is used to modify the amino functional polymer via N‐(3‐dimethylaminopropyl)‐N′‐ethylcarbodiimide hydrochloride/N‐hydroxysuccinimide chemistry for the further steps. The selective adhesion of folate receptor positive cells on the surface is followed by the electrochemical methods. Cyclic voltammetry and electrochemical impedance spectroscopy have been used to characterize stepwise modification of the electroactive surface. After optimization studies such as scan rate during the polymer deposition, FA amount for the efficient surface targeting, incubation time with the cells etc., analytical characterization is carried out. The surface morphologies at each step are imaged by using fluorescence microscopy.

     

Dual‐micropillar‐based microfluidic platform for single embryonic stem cell‐derived neuronal differentiation

We developed the dual‐micropillar‐based microfluidic platform to direct embryonic stem (ES) cell fate. 4 × 4 dual‐micropillar‐based microfluidic platform consisted of 16 circular‐shaped outer micropillars and 8 saddle‐shaped inner micropillars in which single ES cells were cultured. We hypothesized that dual‐micropillar arrays would play an important role in controlling the shear stress and cell docking. Circular‐shaped outer micropillars minimized the shear stress, whereas saddle‐shaped inner micropillars allowed for docking of individual ES cells. We observed the effect of saddle‐shaped inner micropillars on cell docking in response to hydrodynamic resistance. We also demonstrated that ES cells cultured for 6 days within the dual‐micropillar‐based microfluidic platform differentiated into neural‐like cells. Therefore, this dual‐micropillar‐based microfluidic platform could be a potentially powerful method for screening of lineage commitments of single ES cells.     

A Novel BODIPY‐Based Low‐Band‐Gap Small‐Molecule Acceptor for Efficient Non‐fullerene Polymer Solar Cells

We report herein an efficient A¹‐C≡C‐A²‐C≡C‐A¹ type small‐molecule 4,4'‐difluoro‐4‐bora‐3a,4a‐diaza‐s‐ indacene (BODIPY) acceptor (A¹=BODIPY and A²=diketopyrrolopyrrole (DPP)) by following the A‐to‐A excited electron delocalization via the BODIPY meso ‐position, the inherent directionality for the excited electron delocalization. The lowest unoccupied molecular orbital (LUMO) delocalizes across over whole the two flanking A¹ and the central A², and the highest occupied molecular orbital (HOMO) localizes dominantly on the ‐C≡C‐DPP‐C≡C‐ segment. The excited electron upon light excitation of the DPP segment delocalizes over both the BODIPY and DPP segments. The acceptor in chloroform shows an unprecedented plateau‐like broad absorption between 550 and 700 nm with a large FWHM value of 195 nm. Upon transition into solid film, the acceptor shows absorption in the whole near ultraviolet‐visible‐near infrared wavelength region (300‐830 nm) with a low band gap of 1.5 eV and a maximum absorptivity of 0.85×10⁵ cm^‐1. Introduction of the ethynyl spacer between the A¹ and A² and the close BODIPY‐to‐DPP LUMO energy levels are crucial for the excited π−electron delocalization across over whole the conjugation backbone. A power conversion efficiency of 6.60% was obtained from the ternary non‐fullerene solar cell with PTB7‐Th:p ‐DTS(FBTTh₂)₂ (0.5 : 0.5) as the donor materials, which is the highest value among the non‐fullerene organic solar cells with BODIPY as the electron acceptor material.     

Effect of engineered PLGA‐gelatin‐chitosan/PLGA‐gelatin/PLGA‐gelatin‐graphene three‐layer scaffold on adhesion/proliferation of HUVECs

A three‐layered fibrous scaffold composed of fibers of different diameters in each layer was fabricated in correspondence with the structure of the blood vessels. Effect of solution and electrospinning parameters on morphology and diameters of the fibers were investigated by scanning electron microscopy (SEM), for each layer. The SEM images showed that 18% poly (lactic‐co‐glycolic acid) (PLGA)‐gelatin‐chitosan in 1,1,1,3,3,3‐hexafluoro‐2‐propanol (HFIP)/acid acetic solution resulted in bead‐free fibers for the outer layer. For the middle layer, 18% PLGA‐gelatin in HFIP at 13 kV with 13 cm needle to collector distance was chosen as the optimum condition. SEM imaging demonstrated that by increasing graphene content from 0.5 to 2 wt% in the inner layer (as an electrically conductive/platelet anti‐adhesion material), the fiber diameter decreased from 324.01 ± 58.90 to 288.59 ± 70.77 nm. The effect of gelatin crosslinking on the microstructure of the fibers was also examined. Shrinkage ratio decreased from 57 to below 21% upon crosslinking of the three‐layered scaffold in exposure to vapor of 50% glutaraldehyde solution for 2 hours. Mechanical test showed that tensile strength of the crosslinked three‐layer scaffold in the longitudinal direction was 2.90 MPa which is comparable to that of the vein and artery. The MTT [3‐(4,5‐Dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide] assay displayed cell viability of above 96% for the PLGA‐gelatin containing 2 wt% graphene. SEM analysis revealed that the addition of graphene to PLGA‐gelatin (up to 2%) causes a remarkable improvement in cell adhesion.     

Visible‐Light‐Mediated Thiol‐Ene Hydrogelation Using Eosin‐Y as the Only Photoinitiator

The utility of visible‐light‐mediated polymerization in tissue engineering has been limited due to the necessary use of potentially cytotoxic coinitiator and comonomer. Here, we report a visible‐light‐mediated thiol‐ene hydrogelation scheme using eosin‐Y as the only photoinitiator. Under visible light exposure, rapid and highly tunable step‐growth gelation is achieved using PEG‐norbornene and a model cross‐linker dithiothreitol. In addition to investigating the gelation kinetics and properties of thiol‐ene hydrogels formed by this new gelation scheme, we also report high cytocompatibility of these hydrogels using human mesenchymal stem cells (hMSCs) and pancreatic MIN6 β‐cells.     

A Drug‐Free Tumor Therapy Strategy: Cancer‐Cell‐Targeting Calcification

Herein, we propose a drug‐free approach to cancer therapy that involves cancer cell targeting calcification (CCTC). Several types of cancer cells, such as HeLa cells, characterized by folate receptor (FR) overexpression, can selectively adsorb folate (FA) molecules and then concentrate Ca²⁺ locally to induce specific cell calcification. The resultant calcium mineral encapsulates the cancer cells, inducing their death, and in vivo assessments confirm that CCTC treatment can efficiently inhibit tumor growth and metastasis without damaging normal cells compared with conventional chemotherapy. Accordingly, CCTC remarkably improve the survival rate of tumor mice. Notably, both FA and calcium ions are essential ingredients in human metabolism, which means that CCTC is a successful drug‐free method for tumor therapy. This achievement may further represent an alternative cancer therapy characterized by selective calcification‐based substitution of sclerosis for tumor disease.     

Photo‐crosslinkable hydrogel‐based 3D microfluidic culture device

We developed the photo‐crosslinkable hydrogel‐based 3D microfluidic device to culture neural stem cells (NSCs) and tumors. The photo‐crosslinkable gelatin methacrylate (GelMA) polymer was used as a physical barrier in the microfluidic device and collagen type I gel was employed to culture NSCs in a 3D manner. We demonstrated that the pore size was inversely proportional to concentrations of GelMA hydrogels, showing the pore sizes of 5 and 25 w/v% GelMA hydrogels were 34 and 4 μm, respectively. It also revealed that the morphology of pores in 5 w/v% GelMA hydrogels was elliptical shape, whereas we observed circular‐shaped pores in 25 w/v% GelMA hydrogels. To culture NSCs and tumors in the 3D microfluidic device, we investigated the molecular diffusion properties across GelMA hydrogels, indicating that 25 w/v% GelMA hydrogels inhibited the molecular diffusion for 6 days in the 3D microfluidic device. In contrast, the chemicals were diffused in 5 w/v% GelMA hydrogels. Finally, we cultured NSCs and tumors in the hydrogel‐based 3D microfluidic device, showing that 53–75% NSCs differentiated into neurons, while tumors were cultured in the collagen gels. Therefore, this photo‐crosslinkable hydrogel‐based 3D microfluidic culture device could be a potentially powerful tool for regenerative tissue engineering applications.     

Synthesis of 1‐C‐Glycoside‐Linked Lipid II Analogues Toward Bacterial Transglycosylase Inhibition

Preparation of Lipid II analogues containing an enzymatically uncleavable 1‐C‐glycoside linkage between the disaccharide moiety and the pyrophosphate‐ or pyrophosphonate‐lipid moiety is described. The synthesis of a common 1‐C‐vinyl disaccharide intermediate has been developed that allows easy preparation of both an elongated sugar‐phosphate bond and a sugar‐phosphonate moiety, which are coupled with the polyprenyl phosphate to give the desired molecules. Inhibition studies show how a subtle structural modification results in dramatically different potency toward bacterial transglycosylase (TGase), and the results identify Lipid II‐C‐O‐PP (IC₅₀=25 μM ) as a potential TGase inhibitor.     

Effect of Casting Solvent on Morphology and Physical Properties of Partially Sulfonated Polystyrene‐block‐poly(ethylene‐ran‐butylene)‐block‐polystyrene Copolymers

The effect of casting solvent on the morphology of sulfonated polystyrene‐block‐poly(ethylene‐ran‐butylene)‐block‐polystyrene (SSEBS) was investigated using transmission electron microscopy and small‐angle X‐ray scattering. Proton conductivities and methanol permeabilities were measured and the related impact on morphological changes is discussed. SSEBS is transformed from a well‐ordered lamellar to a disordered structure as the concentration of MeOH in MeOH/THF mixtures increases.     

Synthesis of all‐conjugated poly(3‐hexylthiophene)‐block‐ poly(3‐(4′‐(3″,7″‐dimethyloctyloxy)‐3′‐pyridinyl)thiophene) and its blend for photovoltaic applications

New all‐conjugated block copolythiophene, poly(3‐hexylthiophene)‐block‐poly(3‐(4′‐(3″,7″‐dimethyloctyloxy)‐3′‐pyridinyl)thiophene) (P3HT‐b‐P3PyT) was successfully prepared by Grignard metathesis polymerization. The supramolecular interaction between [6,6]‐phenyl‐C₆₁‐butyric acid methyl ester (PCBM) and P3PyT was proposed to control the aggregated size of PCBM and long‐term thermal stability of the photovoltaic cell, as evidenced by differential scanning calorimetry (DSC), transmission electron microscopy (TEM), and optical microscopy. The effect of different solvents on the electronic and optoelectronic properties was studied, including chloroform (CL), dichlorobenzene (DCB), and mixed solvent of CL/DCB. The optimized bulk heterojunction solar cell devices using the P3HT‐b‐P3PyT/PCBM blend showed a power conversion efficiency of 2.12%, comparable to that of P3HT/PCBM device despite the fact that former had a lower crystallinity or absorption coefficient. Furthermore, P3HT‐b‐P3PyT could be also used as a surfactant to enhance the long‐term thermal stability of P3HT/PCBM‐based solar cells by limiting the aggregated size of PCBM. This study represents a new supramolecular approach to design all‐conjugated block copolymers for high‐performance photovoltaic devices. © 2011 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2011.