Kinetics of Activation of Carboxyls to Succinimidyl Ester Groups in Monolayers Grafted on Silicon: An in Situ Real-Time Infrared Spectroscopy Study

Touahir, Larbi; Chazalviel, J.‐N.; Sam, S.; Moraillon, A.; Villeneuve, Catherine Henry de; Allongue, P.; Ozanam, F.; Gouget-Laemmel, Anne Chantal

doi:10.1021/jp200150m

Cited by 17 publications

(22 citation statements)

References 32 publications

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“…Figure 1 demonstrates the corresponding FT-IR spectra of non-modified and modified (5:1, 10:1) fibrinogen and NHS-functionalized PEG. PEG-NHS has a specific succinimidyl ester triplet band (1739, 1782, and 1813 cm −1 ), which was not revealed in the modified fibrinogen samples due to the reaction with primary amino groups caused by imide ringopening [6,7]. These samples had an increasing band at 1103 cm −1 , which indicates the insertion of PEG-derived (C-O) units and varies depending on molar ratio.…”

Section: Resultsmentioning

confidence: 95%

Transparent PEG-Fibrin Gel as a Flexible Tool for Cell Encapsulation

Shpichka¹,

Revkova²,

Aksenova³

et al. 2018

Sovrem Tehnol Med

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The aim of this study was to modify the chemical structure and to optimize the composition of the fibrin gel for effective cell encapsulation. Materials and Methods. We prepared PEGylated fibrin gels using different fibrinogen concentrations (25-50 mg/ml) and PEGfibrinogen molar ratio 10:1 and 5:1 and characterized them via Fourier transform infrared spectroscopy and differential scanning calorimetry. Within the gels, we encapsulated primary culture of fibroblasts and analyzed using light and laser confocal microscopy. Results. PEGylation of fibrinogen allowed us to achieve the gel transparency and preserve its biocompatibility. We revealed that the gel prepared from PEGylated 5:1 fibrinogen (25 mg/ml) provided the most favorable microenvironment for spreading, growth, and proliferation of fibroblasts. This PEG-fibrin gel can be used for encapsulation of different cell types that is essential for various approaches in tissue engineering and diagnostic systems.

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Section: Resultsmentioning

confidence: 95%

Transparent PEG-Fibrin Gel as a Flexible Tool for Cell Encapsulation

Shpichka¹,

Revkova²,

Aksenova³

et al. 2018

Sovrem Tehnol Med

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“…98 The incomplete coverage with NHS esters is probably due to the complex carbodiimide-mediated mechanism of the reaction, which leads to the formation of byproducts during the activation step (N-acylurea) or side products (anhydrides between two adjacent COOH groups), which in both cases could interfere with the formation of new active sites on the surface. 98 However, the presence of NHS functionalities was confirmed by the C 1s, N 1s, and O 1s XPS narrow scans (Supporting Information Figure S1a,c,e). The two O 1s peaks at 532.8 eV (oxygen in carbonyl group) and 535.5 eV (oxygen in C−O−N bonds) are characteristic for ester formation.…”

Section: Resultsmentioning

confidence: 99%

Copper-Free Click Biofunctionalization of Silicon Nitride Surfaces via Strain-Promoted Alkyne–Azide Cycloaddition Reactions

et al. 2012

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Cu-free "click" chemistry is explored on silicon nitride (Si(3)N(4)) surfaces as an effective way for oriented immobilization of biomolecules. An ω-unsaturated ester was grafted onto Si(3)N(4) using UV irradiation. Hydrolysis followed by carbodiimide-mediated activation yielded surface-bound active succinimidyl and pentafluorophenyl ester groups. These reactive surfaces were employed for the attachment of bicyclononyne with an amine spacer, which subsequently enabled room temperature strain-promoted azide-alkyne cycloaddition (SPAAC). This stepwise approach was characterized by means of static water contact angle, X-ray photoelectron spectroscopy, and fluorescence microscopy. The surface-bound SPAAC reaction was studied with both a fluorine-tagged azide and an azide-linked lactose, yielding hydrophobic and bioactive surfaces for which the presence of trace amounts of Cu ions would have been problematic. Additionally, patterning of the Si(3)N(4) surface using this metal-free click reaction with a fluorescent azide is shown. These results demonstrate the ability of the SPAAC as a generic tool for anchoring complex molecules onto a surface under extremely mild, namely ambient and metal-free, conditions in a clean and relatively fast manner.

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“…Figure 1A shows the corresponding FT-IR spectra of pure fibrinogen, NHS-functionalized PEG, and the gels modified with 5:1 and 10:1 molar ratios of PEG to fibrinogen. The PEG-NHS shows a characteristic triplet band of the succinimidyl ester (1739, 1782, and 1813 cm −1 ), which is not present in the modified gels due to imide ring-opening within the reaction with primary amino groups [19,20]. The modified gels show an increasing band at 1103 cm −1 , indicating the insertion of PEG-derived (C-O) units depending on the amount of added NHS-activated PEG.…”

Section: Ft-ir Analysis Of Fibrinogen Pegylationmentioning

confidence: 99%

Hydrogel-based microfluidics for vascular tissue engineering

Koroleva

Deiwick

Nguyen³

et al. 2016

BioNanoMaterials

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Abstract:In this work, we have explored 3-D co-culture of vasculogenic cells within a synthetically modified fibrin hydrogel. Fibrinogen was covalently linked with PEG-NHS in order to improve its degradability resistance and physico-optical properties. We have studied influences of the degree of protein PEGylation and the concentration of enzyme thrombin used for the gel preparation on cellular responses. Scanning electron microscopy analysis of prepared gels revealed that the degree of PEGylation and the concentration of thrombin strongly influenced microstructural characteristics of the protein hydrogel. Human umbilical vein endothelial cells (HUVECs) and human adipose-derived stem cells (hASCs), used as vasculogenic co-culture, could grow in 5:1 PEGylated fibrin gels prepared using 1:0.2 protein to thrombin ratio. This gel formulation supported hASCs and HUVECs spreading and the formation of cell extensions and cell-to-cell contacts. Expression of specific ECM proteins and vasculogenic process inherent cellular enzymatic activity were investigated by immunofluorescent staining, gelatin zymography, western blot and RT-PCR analysis. After evaluation of the optimal gel composition and PEGylation ratio, the hydrogel was utilized for investigation of vascular tube formation within a perfusable microfluidic system. The morphological development of this co-culture within a perfused hydrogel over 12 days led to the formation of interconnected HUVEC-hASC network. The demonstrated PEGylated fibrin microfluidic approach can be used for incorporating other cell types, thus representing a unique experimental platform for basic vascular tissue engineering and drug screening applications.

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Kinetics of Activation of Carboxyls to Succinimidyl Ester Groups in Monolayers Grafted on Silicon: An in Situ Real-Time Infrared Spectroscopy Study

Cited by 17 publications

References 32 publications

Transparent PEG-Fibrin Gel as a Flexible Tool for Cell Encapsulation

Transparent PEG-Fibrin Gel as a Flexible Tool for Cell Encapsulation

Copper-Free Click Biofunctionalization of Silicon Nitride Surfaces via Strain-Promoted Alkyne–Azide Cycloaddition Reactions

Hydrogel-based microfluidics for vascular tissue engineering

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