scholarly journals Localized Enzyme-Assisted Self-Assembly in the Presence of Hyaluronic Acid for Hybrid Supramolecular Hydrogel Coating

Polymers ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 1793
Author(s):  
Jennifer Rodon Fores ◽  
Alexis Bigo-Simon ◽  
Déborah Wagner ◽  
Mathilde Payrastre ◽  
Camille Damestoy ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Self Assembly ◽  
Biologically Active ◽  
Supramolecular Hydrogel ◽  
3T3 Fibroblasts ◽  
Hydrogel Matrix ◽  
Hydrogel Coatings ◽  
Nih 3T3 ◽  
Micrometer Scale ◽  
Hydrogel Coating

Hydrogel coating is highly suitable in biomaterial design. It provides biocompatibility and avoids protein adsorption leading to inflammation and rejection of implants. Moreover, hydrogels can be loaded with biologically active compounds. In this field, hyaluronic acid has been largely studied as an additional component since this polysaccharide is naturally present in extracellular matrix. Strategies to direct hydrogelation processes exclusively from the surface using a fully biocompatible approach are rare. Herein we have applied the concept of localized enzyme-assisted self-assembly to direct supramolecular hydrogels in the presence of HA. Based on electronic and fluorescent confocal microscopy, rheological measurements and cell culture investigations, this work highlights the following aspects: (i) the possibility to control the thickness of peptide-based hydrogels at the micrometer scale (18–41 µm) through the proportion of HA (2, 5 or 10 mg/mL); (ii) the structure of the self-assembled peptide nanofibrous network is affected by the growing amount of HA which induces the collapse of nanofibers leading to large assembled microstructures underpinning the supramolecular hydrogel matrix; (iii) this changing internal architecture induces a decrease of the elastic modulus from 2 to 0.2 kPa when concentration of HA is increasing; (iv) concomitantly, the presence of HA in supramolecular hydrogel coatings is suitable for cell viability and adhesion of NIH 3T3 fibroblasts.

scholarly journals Mechanical Characterization of a Dynamic and Tunable Methacrylated Hyaluronic Acid Hydrogel

2016 ◽  
Vol 138 (2) ◽  
Author(s):  
Matthew G. Ondeck ◽  
Adam J. Engler
Keyword(s):  
Hyaluronic Acid ◽  
Elastic Modulus ◽  
Monomer Concentration ◽  
Degree Of Polymerization ◽  
Force Microscopy ◽  
3T3 Fibroblasts ◽  
Atomic Force ◽  
Spread Area ◽  
Nih 3T3

Hyaluronic acid (HA) is a commonly used natural polymer for cell scaffolding. Modification by methacrylate allows it to be polymerized by free radicals via addition of an initiator, e.g., light-sensitive Irgacure, to form a methacrylated hyaluronic acid (MeHA) hydrogel. Light-activated crosslinking can be used to control the degree of polymerization, and sequential polymerization steps allow cells plated onto or in the hydrogel to initially feel a soft and then a stiff matrix. Here, the elastic modulus of MeHA hydrogels was systematically analyzed by atomic force microscopy (AFM) for a number of variables including duration of UV exposure, monomer concentration, and methacrylate functionalization. To determine how cells would respond to a specific two-step polymerization, NIH 3T3 fibroblasts were cultured on the stiffening MeHA hydrogels and found to reorganize their cytoskeleton and spread area upon hydrogel stiffening, consistent with cells originally cultured on substrates of the final elastic modulus.

scholarly journals Posttranscriptional control of human gamma interferon gene expression in transfected mouse fibroblasts.

1986 ◽  
Vol 6 (6) ◽  
pp. 2253-2256 ◽  
Author(s):  
H A Young ◽  
L Varesio ◽  
P Hwu
Keyword(s):  
Gene Expression ◽  
Genomic Dna ◽  
Gamma Interferon ◽  
Biologically Active ◽  
Posttranscriptional Control ◽  
Transfected Cells ◽  
3T3 Fibroblasts ◽  
Nih 3T3 ◽  
Interferon Gene ◽  
Calcium Phosphate Precipitation

Human gamma interferon genomic DNA was introduced into NIH 3T3 fibroblasts by calcium phosphate precipitation and was not expressed in these cells at the cytoplasmic mRNA or protein level. Treatment of the transfected cells with cycloheximide (1 microgram/ml) induced the accumulation of cytoplasmic gamma interferon mRNA and biologically active human gamma interferon. Analysis of the nuclear enriched RNA from untreated cells indicated that human gamma interferon mRNA was present, suggesting that cycloheximide may act by inhibiting a specific nuclease or may enhance the processing or transport of the RNA from the nucleus to the cytoplasm.

Posttranscriptional control of human gamma interferon gene expression in transfected mouse fibroblasts

1986 ◽  
Vol 6 (6) ◽  
pp. 2253-2256
Author(s):  
H A Young ◽  
L Varesio ◽  
P Hwu
Keyword(s):  
Gene Expression ◽  
Genomic Dna ◽  
Gamma Interferon ◽  
Biologically Active ◽  
Posttranscriptional Control ◽  
Transfected Cells ◽  
3T3 Fibroblasts ◽  
Nih 3T3 ◽  
Interferon Gene ◽  
Calcium Phosphate Precipitation

Human gamma interferon genomic DNA was introduced into NIH 3T3 fibroblasts by calcium phosphate precipitation and was not expressed in these cells at the cytoplasmic mRNA or protein level. Treatment of the transfected cells with cycloheximide (1 microgram/ml) induced the accumulation of cytoplasmic gamma interferon mRNA and biologically active human gamma interferon. Analysis of the nuclear enriched RNA from untreated cells indicated that human gamma interferon mRNA was present, suggesting that cycloheximide may act by inhibiting a specific nuclease or may enhance the processing or transport of the RNA from the nucleus to the cytoplasm.

scholarly journals Differential processing of colony-stimulating factor 1 precursors encoded by two human cDNAs.

1988 ◽  
Vol 8 (11) ◽  
pp. 5026-5034 ◽  
Author(s):  
C W Rettenmier ◽  
M F Roussel
Keyword(s):  
Amino Acid ◽  
Growth Factor ◽  
Biologically Active ◽  
Colony Stimulating Factor ◽  
Acid Product ◽  
Human Cdna ◽  
3T3 Fibroblasts ◽  
Nih 3T3 ◽  
Stimulating Factor

The biosynthesis of macrophage colony-stimulating factor 1 (CSF-1) was examined in mouse NIH-3T3 fibroblasts transfected with a retroviral vector expressing the 554-amino-acid product of a human 4-kilobase (kb) CSF-1 cDNA. Similar to results previously obtained with a 1.6-kb human cDNA that codes for a 256-amino-acid CSF-1 precursor, the results of the present study showed that NIH-3T3 cells expressing the product of the 4-kb clone produced biologically active human CSF-1 and were transformed by an autocrine mechanism when cotransfected with a vector containing a human c-fms (CSF-1 receptor) cDNA. The 4-kb CSF-1 cDNA product was synthesized as an integral transmembrane glycoprotein that was assembled into disulfide-linked dimers and rapidly underwent proteolytic cleavage to generate a soluble growth factor. Although the smaller CSF-1 precursor specified by the 1.6-kb human cDNA was stably expressed as a membrane-bound glycoprotein at the cell surface and was slowly cleaved to release the extracellular growth factor, the cell-associated product of the 4-kb clone was efficiently processed to the secreted form and was not detected on the plasma membrane. Digestion with glycosidic enzymes indicated that soluble CSF-1 encoded by the 4-kb cDNA contained both asparagine(N)-linked and O-linked carbohydrate chains, whereas the product of the 1.6-kb clone had only N-linked oligosaccharides. Removal of the carbohydrate indicated that the polypeptide chain of the secreted 4-kb cDNA product was longer than that of the corresponding form encoded by the smaller clone. These differences in posttranslational processing may reflect diverse physiological roles for the products of the two CSF-1 precursors in vivo.

Differential processing of colony-stimulating factor 1 precursors encoded by two human cDNAs

1988 ◽  
Vol 8 (11) ◽  
pp. 5026-5034
Author(s):  
C W Rettenmier ◽  
M F Roussel
Keyword(s):  
Amino Acid ◽  
Growth Factor ◽  
Biologically Active ◽  
Colony Stimulating Factor ◽  
Acid Product ◽  
Human Cdna ◽  
3T3 Fibroblasts ◽  
Nih 3T3 ◽  
Stimulating Factor

The biosynthesis of macrophage colony-stimulating factor 1 (CSF-1) was examined in mouse NIH-3T3 fibroblasts transfected with a retroviral vector expressing the 554-amino-acid product of a human 4-kilobase (kb) CSF-1 cDNA. Similar to results previously obtained with a 1.6-kb human cDNA that codes for a 256-amino-acid CSF-1 precursor, the results of the present study showed that NIH-3T3 cells expressing the product of the 4-kb clone produced biologically active human CSF-1 and were transformed by an autocrine mechanism when cotransfected with a vector containing a human c-fms (CSF-1 receptor) cDNA. The 4-kb CSF-1 cDNA product was synthesized as an integral transmembrane glycoprotein that was assembled into disulfide-linked dimers and rapidly underwent proteolytic cleavage to generate a soluble growth factor. Although the smaller CSF-1 precursor specified by the 1.6-kb human cDNA was stably expressed as a membrane-bound glycoprotein at the cell surface and was slowly cleaved to release the extracellular growth factor, the cell-associated product of the 4-kb clone was efficiently processed to the secreted form and was not detected on the plasma membrane. Digestion with glycosidic enzymes indicated that soluble CSF-1 encoded by the 4-kb cDNA contained both asparagine(N)-linked and O-linked carbohydrate chains, whereas the product of the 1.6-kb clone had only N-linked oligosaccharides. Removal of the carbohydrate indicated that the polypeptide chain of the secreted 4-kb cDNA product was longer than that of the corresponding form encoded by the smaller clone. These differences in posttranslational processing may reflect diverse physiological roles for the products of the two CSF-1 precursors in vivo.

scholarly journals Evaluation of a Model Photo-Caged Dehydropeptide as a Stimuli-Responsive Supramolecular Hydrogel

Nanomaterials ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 704
Author(s):  
Peter J. Jervis ◽  
Loic Hilliou ◽  
Renato B. Pereira ◽  
David M. Pereira ◽  
José A. Martins ◽  
...  
Keyword(s):  
Self Assembly ◽  
Uv Light ◽  
Aqueous Media ◽  
Intramolecular Interactions ◽  
Biologically Active ◽  
Wound Dressings ◽  
Supramolecular Hydrogel ◽  
On Demand ◽  
Proteolytic Resistance

Short peptides capped on the N-terminus with aromatic groups are often able to form supramolecular hydrogels, via self-assembly, in aqueous media. The rheological properties of these readily tunable hydrogels resemble those of the extracellular matrix (ECM) and therefore have potential for various biological applications, such as tissue engineering, biosensors, 3D bioprinting, drug delivery systems and wound dressings. We herein report a new photo-responsive supramolecular hydrogel based on a “caged” dehydropeptide (CNB-Phe-ΔPhe-OH 2), containing a photo-cleavable carboxy-2-nitrobenzyl (CNB) group. We have characterized this hydrogel using a range of techniques. Irradiation with UV light cleaves the pendant aromatic capping group, to liberate the corresponding uncaged model dehydropeptide (H-Phe-ΔPhe-OH 3), a process which was investigated by 1H NMR and HPLC studies. Crucially, this cleavage of the capping group is accompanied by dissolution of the hydrogel (studied visually and by fluorescence spectroscopy), as the delicate balance of intramolecular interactions within the hydrogel structure is disrupted. Hydrogels which can be disassembled non-invasively with temporal and spatial control have great potential for specialized on-demand drug release systems, wound dressing materials and various topical treatments. Both 2 and 3 were found to be non-cytotoxic to the human keratinocyte cell line, HaCaT. The UV-responsive hydrogel system reported here is complementary to previously reported related UV-responsive systems, which are generally composed of peptides formed from canonical amino acids, which are susceptible to enzymatic proteolysis in vivo. This system is based on a dehydrodipeptide structure which is known to confer proteolytic resistance. We have investigated the ability of the photo-activated system to accelerate the release of the antibiotic, ciprofloxacin, as well as some other small model drug compounds. We have also conducted some initial studies towards skin-related applications. Moreover, this model system could potentially be adapted for on-demand “self-delivery”, through the uncaging of known biologically active dehydrodipeptides.

Layer-by-Layer Self-Assembly of Carbon Nanotubes with Polyelectrolytes for Biomedical Application

2013 ◽  
Vol 749 ◽  
pp. 359-361
Author(s):  
Lin Su ◽  
Pei Pei Zhao ◽  
Yun Feng Shi ◽  
Li Ting Yao ◽  
Wei Xue ◽  
...  
Keyword(s):  
Carbon Nanotube ◽  
Self Assembly ◽  
Biomedical Application ◽  
Layer By Layer ◽  
3T3 Fibroblasts ◽  
Multi Walled Carbon Nanotube ◽  
Biomedical Field ◽  
Cell Biocompatibility ◽  
Nih 3T3 ◽  
Positively Charged

The layer-by-layer (LbL) self-assembly of carbon nanotube (CNT)/polyelectrolyte has been widely investigated for various applications. In this study, multi-walled carbon nanotube (MWCNT) was LbL assembled with three positively charged polyelectrolytes, which was PEI, chitosan and PDDA respectively. In order to investigate its potential application in biomedical field, cell biocompatibility was examined by examining the viability and morphologies of NIH-3T3 fibroblasts cultured on the glass coated with the MWCNT/polyelectrolyte assembly multi-layers.

scholarly journals Self-Targeting of Carbon Dots into the Cell Nucleus: Diverse Mechanisms of Toxicity in NIH/3T3 and L929 Cells

2021 ◽  
Vol 22 (11) ◽  
pp. 5608
Author(s):  
Markéta Havrdová ◽  
Iztok Urbančič ◽  
Kateřina Bartoň Tománková ◽  
Lukáš Malina ◽  
Janez Štrancar ◽  
...  
Keyword(s):  
Dna Damage ◽  
Carbon Dots ◽  
L929 Cell ◽  
Fluorescent Properties ◽  
Cell Cycle Profile ◽  
Mechanisms Of Toxicity ◽  
3T3 Fibroblasts ◽  
Nih 3T3 ◽  
Positively Charged ◽  
Endocytic Pathways

It is important to understand the nanomaterials intracellular trafficking and distribution and investigate their targeting into the nuclear area in the living cells. In our previous study, we firstly observed penetration of nonmodified positively charged carbon dots decorated with quaternary ammonium groups (QCDs) into the nucleus of mouse NIH/3T3 fibroblasts. Thus, in this work, we focused on deeper study of QCDs distribution inside two healthy mouse NIH/3T3 and L929 cell lines by fluorescence microspectroscopy and performed a comprehensive cytotoxic and DNA damage measurements. Real-time penetration of QCDs across the plasma cell membrane was recorded, concentration dependent uptake was determined and endocytic pathways were characterized. We found out that the QCDs concentration of 200 µg/mL is close to saturation and subsequently, NIH/3T3 had a different cell cycle profile, however, no significant changes in viability (not even in the case with QCDs in the nuclei) and DNA damage. In the case of L929, the presence of QCDs in the nucleus evoked a cellular death. Intranuclear environment of NIH/3T3 cells affected fluorescent properties of QCDs and evoked fluorescence blue shifts. Studying the intracellular interactions with CDs is essential for development of future applications such as DNA sensing, because CDs as DNA probes have not yet been developed.

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