Modulation of Living Cell Behavior with Ultra‐Low Fouling Polymer Brush Interfaces

Víšová, Ivana; Smolková, Barbora; Uzhytchak, Mariia; Vrabcová, Markéta; Zhigunová, Yulia; Houška, Milan; Surman, František; Pereira, Andres los Santos; Lunov, Oleg; Dejneka, A.; Vaisocherová‐Lísalová, Hana

doi:10.1002/mabi.202070005

Cited by 6 publications

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“…A set of pCB brush coatings and OEG-based SAM coatings functionalized with variable concentrations of RGD peptides were prepared to investigate the effects of the RGD peptide-decorated antifouling background on mechanotransduction cell signaling. Based on our previous study [ 31 ], we used optimized biocompatible and highly hydratable antifouling pCB brush coatings with wet thicknesses around ~80 nm (characterized by spectroscopic ellipsometry; data not shown). Two different approaches to the preparation of the coatings functionalized with varying RGD concentrations were employed as follows: first, only carboxy-functional coatings of pCBAA or SAM of HS-(CH 2 ) 11 -(EG) 6 -OCH 2 -COOH were RGD-functionalized using immobilization solutions of RGD peptides of different concentrations ( Figure 1 A1,A2).…”

Section: Resultsmentioning

confidence: 99%

“…Hepatocytes are very sensitive cells [ 35 , 36 , 37 ], which makes them a good model system for mechanotransduction-signaling studies. Recently we showed that nonfunctionalized antifouling polymer brush coatings pCBAA, pCBMAA, pHPMAA, and their copolymers are not cytotoxic for Huh7 cells [ 31 ]. Here, we confirmed no cytotoxicity, even for activated and deactivated or RGD-functionalized pCB antifouling coatings.…”

Section: Resultsmentioning

confidence: 99%

“…However, even the RGD-functionalized homopolymer of 100% pCBMAA (functionalizable part of the copolymer brushes) did not reach the level of YAP colocalization induced by the control or the RGD-functionalized homopolymer of pCBAA ( Figure 3 B). In our previous study, the sensitivity of cell growth and spreading on the surface swelling characteristics was noticed, causing the higher growth and cytoskeleton distribution of cells growing on the nonfunctionalized pCBAA compared to pCBMAA [ 31 ]. Such a phenomenon can explain the different responses of the cells on functionalized pCBAA (method A-1 in Figure 1 ) and pCBMAA (method B-1 in Figure 1 ) and opens new possibilities in the design of coatings for the even finer modulation of cell growth and cell-signaling pathways.…”

Section: Resultsmentioning

confidence: 99%

“…pCB brushes were prepared by surface-initiated atom transfer radical polymerization (SI-ATRP), as described elsewhere [ 31 ], and their structures were checked by infrared spectroscopy (data not shown). After polymerization, all coatings were stored in PBS.…”

Section: Methodsmentioning

confidence: 99%

“…To define the level of mechanical stress, we monitored the colocalization of YAP in the cell nucleus. Two distinct types of state-of-art functionable antifouling coatings based on pCB zwitterionic brushes [ 29 , 30 , 31 ] were employed and compared. First, a set of RGD-functionalized homopolymers poly(carboxybetaine acrylamide) (pCBAA) was prepared by employing different concentrations of the RGD peptide during the immobilization.…”

Section: Introductionmentioning

confidence: 99%

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Functionalizable Antifouling Coatings as Tunable Platforms for the Stress-Driven Manipulation of Living Cell Machinery

et al. 2020

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Cells are continuously sensing their microenvironment and subsequently respond to different physicochemical cues by the activation or inhibition of different signaling pathways. To study a very complex cellular response, it is necessary to diminish background environmental influences and highlight the particular event. However, surface-driven nonspecific interactions of the abundant biomolecules from the environment influence the targeted cell response significantly. Yes-associated protein (YAP) translocation may serve as a marker of human hepatocellular carcinoma (Huh7) cell responses to the extracellular matrix and surface-mediated stresses. Here, we propose a platform of tunable functionable antifouling poly(carboxybetain) (pCB)-based brushes to achieve a molecularly clean background for studying arginine, glycine, and aspartic acid (RGD)-induced YAP-connected mechanotransduction. Using two different sets of RGD-functionalized zwitterionic antifouling coatings with varying compositions of the antifouling layer, a clear correlation of YAP distribution with RGD functionalization concentrations was observed. On the other hand, commonly used surface passivation by the oligo(ethylene glycol)-based self-assembled monolayer (SAM) shows no potential to induce dependency of the YAP distribution on RGD concentrations. The results indicate that the antifouling background is a crucial component of surface-based cellular response studies, and pCB-based zwitterionic antifouling brush architectures may serve as a potential next-generation easily functionable surface platform for the monitoring and quantification of cellular processes.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Functionalizable Antifouling Coatings as Tunable Platforms for the Stress-Driven Manipulation of Living Cell Machinery

et al. 2020

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Tuning of Surface Charge of Functionalized Poly(Carboxybetaine) Brushes Can Significantly Improve Label‐Free Biosensing in Complex Media

Víšová

Houska

Spasovová

et al. 2022

Adv Materials Inter

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Besides the resistance to fouling, the platform should be easily functionalized, i.e., conjugated with molecules having specific biological activity, usually a high affinity for specific targets. [9][10][11][12][13][14][15][16] Applications that require such functionalized antifouling platforms span from rapid detection of chemical and biological species, coatings of nanoparticles used in drug delivery, membranes for separation and cleaning technologies, to scaffolds for tissue engineering.Poly(carboxybetaine) (pCB) brushes are outstanding antifouling platforms allowing facile functionalization with biorecognition elements (BREs) via EDC/NHS (carbodiimide/N-hydroxysuccinimide) chemistry. [17][18][19] Their extraordinary properties stem from their high hydrophilicity and overall electroneutrality, which makes them resistant to hydrophobic as well as electrostatic adsorption from contacted media. However, their net electric charge is pH dependent. The positive charge of quaternary ammonium group is permanent but pKa of pCB carboxyl group is somewhere between 2 and 4. [17,20,21] The measurements of zeta potential of the surface bound poly(carboxybetaine methacrylamide) (pCBMAA), [22] indicated that its isoelectric point (pI) is around 8.5 and thus it is positively charged at lower pH values. The net positive charge of functionalized pCB brushes is further enhanced due to the consumption of betaine carboxyl groups following BRE conjugation reaction (Figure 1). It should be taken into consideration that conjugated BRE may also induce charge shifts and thus an effective optimization of the platform surface charge balance is of importance. [16] The overall reaction scheme for the BRE conjugation via EDC/NHS is shown in Figure 1. The carboxyl group of pCB is converted to active NHS ester which readily reacts with amino group of BRE to create stable amide bond. However, not all NHS esters buried inside pCB brushes are able to react with bulky BREs that cannot penetrate below the surface, but can react nonspecifically with other smaller amino compounds present in complex biological media. Therefore, all residual NHS esters must be eliminated (deactivated) as otherwise they could Poly(carboxybetaine) brushes are excellent antifouling platforms allowing facile functionalization with biorecognition elements via carbodiimide/N-hydroxysuccinimide (EDC/NHS) chemistry. However, residual active NHS esters and the loss of zwitterionic balance after the conjugation may impair initially excellent antifouling properties. This problem has so far been addressed either by using spontaneous hydrolysis or deactivation of residual NHS esters by the reaction with a small amino compound bearing hydroxyl or carboxyl groups. In contrast to this approach, and instead of using a single deactivator, here the use of tailored mixtures of deactivating agents containing carboxyl groups and sulfo or sulfate groups with permanent negative charge that allow to tune surface charge balance is investigated. The approach is applied to poly(carboxybetaine acr...

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Synthetic Evolution of a Supramolecular Harpooning Mechanism to Immobilize Vesicles at Antifouling Interfaces

Englert,

Witzdam,

Söder

et al. 2023

Macro Chemistry & Physics

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The immobilization of vesicles has been conceptualized as a method to functionalize biointerfaces. However, the preservation of their integrity post immobilization remains a considerable challenge. Interfacial interactions can cause vesicle rupture upon close surface contact and non‐specific protein adsorption impairing surface functions. To date, immobilization of vesicles has relied solely on either entrapment or prior modification of vesicles, both of which require laborious preparation and limit their applications. In this work, we develop a bioinspired strategy to pin vesicles without prior modification while preserving their intact shape. We introduce antifouling diblock copolymers and ultrathin surface‐attached hydrogels containing a brush‐like interface consisting of a bottle brush copolymer of N‐(2‐hydroxypropyl) methacrylamide (HPMA) and N‐(3‐methacrylamidopropyl)‐N,N‐dimethyldodecan‐1‐aminiumiodide (C12+). The presence of positive charges generates an attractive force that pulls vesicles toward the surface. At the surface, the amphiphilic properties of the combs facilitate their insertion into the membrane, mimicking the harpooning mechanism observed in antimicrobial peptides. Importantly, the antifouling poly(HPMA) backdrop serves to safeguard the vesicles by preventing deformation and breakage. Using a combination of thermodynamic analysis, surface plasmon resonance, and confocal laser scanning microscopy, we demonstrate the efficiency of this biomimetic system to capture vesicles while maintaining an antifouling interface necessary for bioapplications.This article is protected by copyright. All rights reserved

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Modulation of Living Cell Behavior with Ultra‐Low Fouling Polymer Brush Interfaces

Cited by 6 publications

References 0 publications

Functionalizable Antifouling Coatings as Tunable Platforms for the Stress-Driven Manipulation of Living Cell Machinery

Functionalizable Antifouling Coatings as Tunable Platforms for the Stress-Driven Manipulation of Living Cell Machinery

Tuning of Surface Charge of Functionalized Poly(Carboxybetaine) Brushes Can Significantly Improve Label‐Free Biosensing in Complex Media

Synthetic Evolution of a Supramolecular Harpooning Mechanism to Immobilize Vesicles at Antifouling Interfaces

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