Design of bFGF-tethered self-assembling extracellular matrix proteins via coiled-coil triple-helix formation

Mizuguchi, Yoshinori; Mashimo, Yasumasa; Mie, Masayasu; Kobatake, Eiry

doi:10.1088/1748-605x/aa7616

Cited by 4 publications

(6 citation statements)

References 41 publications

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“…Mizuguchi et al 209 developed a heterodimer triple-helix self-assembling ECM comprised of an elastin-derived structural unit (APGVGV)12 and an RGD sequence. They showed that basic fibroblast growth factor (bFGF) can be tethered to this hydrogel using the complimentary triple-helix formation, which increased the stability and activity of immobilized bFGF compared with soluble bFGF and enhanced the proliferation of Human umbilical vein endothelial cells (HUVECs) cultured on the artificial ECM.…”

Section: Other Biofunctionalitiesmentioning

confidence: 99%

Modular Enzyme-Responsive Polysaccharide-Based Hydrogels for Biofabrication

Naeimipour

2023

Linköping Studies in Science and Technology. Dissertations

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Engineered human tissue and disease models can decrease the cost and time of developing new drugs and treatments, facilitate personalized medicine, and eliminate the need for animal models that poorly represent the human body and are ethically problematic. However, the current conventional cell expansion methods using 2D culture flasks cannot enable the development of such complex multi-cellular 3D models. In general, hydrogels are considered promising materials that can make the biofabrication of tissue models possible. Hydrogels are highly hydrated materials comprised of either synthetic or naturally derived polymers, or a combination of both, and can form an environment mimicking the biomacromolecular network surrounding cells in the body. This network of biopolymers, known as extracellular matrix (ECM), is comprised of proteins such as collagen, laminin, fibronectin, and polysaccharides such as hyaluronan (HA), heparan, keratan, and chondroitin sulfate. The design of hydrogels representing the physical and biochemical properties of the ECM and which can be used for biofabrication is challenging but of increasing interest due to the rapid progress in the development of 3D and 4D bioprinting techniques. As the ECM properties differ between various tissues and disease conditions and change over time, a dynamic modular hydrogel system is needed to that can be optimized for each cell and tissue type. This thesis aims to develop modular enzyme-responsive polysaccharide-based hydrogels for 3D cell culture and biofabrication. The natural polysaccharides, hyaluronic acid (HA) and alginate (Alg) were used as the main backbone in the hydrogels developed in this thesis. HA was modified by conjugating bicyclo[6.1.0]non-4-yne (BCN) to the backbone to form HA-BCN-based hydrogels by a bioorthogonal strain-promoted alkyne-azide cycloaddition. The click reaction between BCN and azide groups allowed for modulating the biochemical and mechanical properties of the HA-BCN hydrogels. HA-BCN was further decorated with peptides to explore peptide folding and dimerizationmediated dynamic cross-linking and biofunctionalization. This system was further used to explore possibilities to dynamically alter the properties of 3D bioprinted structures, mimicking the biomineralization process in bone tissue. In a different study, a tumor model comprising fibroblast and breast cancer cells (MCF7) was bioprinted using HA-BCN cross-linked by matrix metalloporotease (MMP) cleavable and PEG-diazide MMP-resistant crosslinkers, demonstrating the synergistic relationship between hydrogel degradability and cancer cell growth, intensified by the presence of fibroblasts. The possibility of incorporating a conductive module into this hydrogel system was explored using the enzyme-assisted polymerization of ETE-S to form an

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Section: Other Biofunctionalitiesmentioning

confidence: 99%

Modular Enzyme-Responsive Polysaccharide-Based Hydrogels for Biofabrication

Naeimipour

2023

Linköping Studies in Science and Technology. Dissertations

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“… 504 As an alternative, pendant helical groups can be attached to a covalent material and used to drive the noncovalent attachment of bioactive groups such as growth factors via self-assembly into coiled-coil triple helices. 505 …”

Section: Noncovalent Systemsmentioning

confidence: 99%

“…This is most easily achieved by mixing a self-assembling peptide into a covalent hydrogel, composed of either a noninteracting polymer such as interpenetrating networks of PEG or systems where additional charge interactions further stabilize the final construct, for example between positively charged peptides and negatively charged alginate gels . As an alternative, pendant helical groups can be attached to a covalent material and used to drive the noncovalent attachment of bioactive groups such as growth factors via self-assembly into coiled-coil triple helices …”

Section: Noncovalent Systemsmentioning

confidence: 99%

Achieving Controlled Biomolecule–Biomaterial Conjugation

2018

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The conjugation of biomolecules can impart materials with the bioactivity necessary to modulate specific cell behaviors. While the biological roles of particular polypeptide, oligonucleotide, and glycan structures have been extensively reviewed, along with the influence of attachment on material structure and function, the key role played by the conjugation strategy in determining activity is often overlooked. In this review, we focus on the chemistry of biomolecule conjugation and provide a comprehensive overview of the key strategies for achieving controlled biomaterial functionalization. No universal method exists to provide optimal attachment, and here we will discuss both the relative advantages and disadvantages of each technique. In doing so, we highlight the importance of carefully considering the impact and suitability of a particular technique during biomaterial design.

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“…The commonly used neurosphere culture method for expanding neural stem cells has the disadvantage of producing a mixture of cell types such as neurons and astrocytes. Currently, surface modification with chimeric proteins is being used to develop functional biomaterials worldwide . Previously, a substrate immobilized with chimeric epidermal growth factor (EGF), which acts as a mitogen for neural stem cells and NSPCs, was designed for the uniform expansion of cells expressing only nestin, a marker of neural stem cell .…”

Section: Introductionmentioning

confidence: 99%

“…Currently, surface modification with chimeric proteins is being used to develop functional biomaterials worldwide. 13,14 Previously, a substrate immobilized with chimeric epidermal growth factor (EGF), which acts as a mitogen for neural stem cells and NSPCs, was designed for the uniform expansion of cells expressing only nestin, a marker of neural stem cell. [15][16][17][18] Therefore, we developed a novel culture method, which utilizes an EGF chimeric protein-anchored substrate to obtain highly pure NSPCs.…”

Section: Introductionmentioning

confidence: 99%

Functional surfaces for efficient differentiation of neural stem/progenitor cells into dopaminergic neurons

Nakaji-Hirabayashi

Fujimoto

Yoshikawa

et al. 2019

J Biomedical Materials Res

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Various techniques and systems have been reported for the efficient differentiation of neural stem/progenitor cells into dopaminergic neurons. Although a comparatively high percentage of dopaminergic neurons can be obtained using these techniques, the differentiated cells display varied cellular phenotypes such as astrocytes and oligodendrocytes. Generation of highly pure dopaminergic neurons is important for cell‐based therapy and in vitro evaluation of dopaminergic neuron function. In this study, we developed a culture surface anchored with several neurotrophic factors and a neuronal cell‐adhesive protein for efficient differentiation of neural stem/progenitor cells into dopaminergic neurons. Oligohistidine‐fused brain‐derived neurotrophic factor and glial cell line–derived neurotrophic factor, synthesized using genetic engineering, were co‐immobilized on the surface via metal chelation. To facilitate cell adhesion, a cell‐adhesive chimeric protein derived from laminin (LN‐G) was also immobilized on the surface. Approximately 40% of the cells cultured for 14 days with these protein‐immobilized substrates expressed tyrosine hydroxylase, a marker of dopaminergic neurons, with a three‐fold increase in differentiation efficiency than that reported previously. In addition, the number of tyrosine hydroxylase‐positive cells increased to approximately 80% of the culture after 30 days. These cells secreted dopamine and expressed dopaminergic neuron‐specific genes. Interestingly, cell types (glial cells and oligodendrocytes) other than neuronal cells (immature and mature dopaminergic neurons) were not detected on the protein‐anchored surface. Our results demonstrate that highly pure dopaminergic neurons can be exclusively obtained using the novel substrate without extra purification steps such as cell sorting. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 860–871, 2019.

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Design of bFGF-tethered self-assembling extracellular matrix proteins via coiled-coil triple-helix formation

Cited by 4 publications

References 41 publications

Modular Enzyme-Responsive Polysaccharide-Based Hydrogels for Biofabrication

Modular Enzyme-Responsive Polysaccharide-Based Hydrogels for Biofabrication

Achieving Controlled Biomolecule–Biomaterial Conjugation

Functional surfaces for efficient differentiation of neural stem/progenitor cells into dopaminergic neurons

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