Recombinant human elastin polypeptides self‐assemble into biomaterials with elastin‐like properties

Bellingham, Catherine M.; Lillie, Margo A.; Gosline, John M.; Wright, Glenda M.; Starcher, Barry; Bailey, Allen J.; Woodhouse, Kimberly A.; Keeley, Fred W.

doi:10.1002/bip.10512

Cited by 235 publications

(232 citation statements)

References 34 publications

(53 reference statements)

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“…Elastin is an insoluble, polymeric, ECM protein that provides various tissues in the body with the properties of extensibility and elastic recoil. Elastin is synthesized as tropoelastin (~70kDa), a soluble precursor to insoluble elastin [30]. Tropoelastin has a repeating domain structure, alternating between hydrophobic and crosslinking regions.…”

Section: Biomimetic Mechanical Propertiesmentioning

confidence: 99%

Biomimetic materials for tissue engineering

Peter

2008

Advanced Drug Delivery Reviews

1,201

819

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Tissue engineering and regenerative medicine is an exciting research area that aims at regenerative alternatives to harvested tissues for transplantation. Biomaterials play a pivotal role as scaffolds to provide three-dimensional templates and synthetic extracellular-matrix environments for tissue regeneration. It is often beneficial for the scaffolds to mimic certain advantageous characteristics of the natural extracellular matrix, or developmental or would healing programs. This article reviews current biomimetic materials approaches in tissue engineering. These include synthesis to achieve certain compositions or properties similar to those of the extracellular matrix, novel processing technologies to achieve structural features mimicking the extracellular matrix on various levels, approaches to emulate cell-extracellular matrix interactions, and biologic delivery strategies to recapitulate a signaling cascade or developmental/would-healing program. The article also provides examples of enhanced cellular/tissue functions and regenerative outcomes, demonstrating the excitement and significance of the biomimetic materials for tissue engineering and regeneration.

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Section: Biomimetic Mechanical Propertiesmentioning

confidence: 99%

Biomimetic materials for tissue engineering

Peter

2008

Advanced Drug Delivery Reviews

1,201

819

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“…We, and others, have shown that elastin-like polypeptides (ELPs) derived from the native sequence of elastin act as suitable proxies to study phase separation and elasticity (18,28,31,39). In the present study, we highlight the design and characterization of an elastin mimetic that undergoes phase separation and can be cross-linked to create resilient biomaterials whose assembly can be monitored by NMR to obtain site-specific resolution of structure and dynamics of both the phase-separated and cross-linked materials.…”

Section: Significancementioning

confidence: 99%

“…Conversely, simple coacervation is considered an entropically driven process that depends entirely on hydrophobic interactions between protein chains (4, 16), although few molecular details of this process have been reported. Thus, a detailed understanding of protein structure and dynamics during LLPS is needed to better define the principles governing protein phase separation and to further develop applications of this type of self-assembly in biomaterial design.The elastin monomer, tropoelastin, undergoes a wellcharacterized lower critical solution temperature (LCST) phase transition that is promoted by the presence of salt and heat (8,17,18). Tropoelastin is an IDP consisting of an alternating arrangement of hydrophobic domains (HDs) and cross-linking domains (CLDs) (7,19).…”

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confidence: 99%

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Direct observation of structure and dynamics during phase separation of an elastomeric protein

Reichheld

Muiznieks

Keeley

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

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212

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Despite its growing importance in biology and in biomaterials development, liquid-liquid phase separation of proteins remains poorly understood. In particular, the molecular mechanisms underlying simple coacervation of proteins, such as the extracellular matrix protein elastin, have not been reported. Coacervation of the elastin monomer, tropoelastin, in response to heat and salt is a critical step in the assembly of elastic fibers in vivo, preceding chemical crosslinking. Elastin-like polypeptides (ELPs) derived from the tropoelastin sequence have been shown to undergo a similar phase separation, allowing formation of biomaterials that closely mimic the material properties of native elastin. We have used NMR spectroscopy to obtain site-specific structure and dynamics of a self-assembling elastin-like polypeptide along its entire self-assembly pathway, from monomer through coacervation and into a cross-linked elastic material. Our data reveal that elastin-like hydrophobic domains are composed of transient β-turns in a highly dynamic and disordered chain, and that this disorder is retained both after phase separation and in elastic materials. Cross-linking domains are also highly disordered in monomeric and coacervated ELP 3 and form stable helices only after chemical cross-linking. Detailed structural analysis combined with dynamic measurements from NMR relaxation and diffusion data provides direct evidence for an entropy-driven mechanism of simple coacervation of a protein in which transient and nonspecific intermolecular hydrophobic contacts are formed by disordered chains, whereas bulk water and salt are excluded.phase separation | elastin | NMR | protein structure | dynamics T he liquid-liquid phase separation (LLPS) of molecules has long been exploited to concentrate and encapsulate molecules for drug delivery and food preparation (1, 2). In biology, there is increasing awareness that many proteins exhibit this type of phase behavior, allowing transient microenvironments to be quickly assembled and disassembled in response to changing solution conditions, or the availability of binding partners (3, 4). Protein phase separation occurs intracellularly to generate various membraneless organelles, such as ribonucleoprotein (RNP) bodies involved in nucleic acid processing, transport, and storage (5). A similar phenomenon is observed in the extracellular matrix as a critical step in the synthesis of elastic fibers, which provide extensibility, recoil, and resilience to tissues (6, 7). In the latter system, LLPS of monomeric elastin results in hydrated protein-rich coacervate droplets that are deposited and crosslinked to form an elastic matrix (7,8). In addition to their fundamental importance to biology, the dynamic reversibility of LLPS makes phase-separated states of proteins an attractive platform for development of responsive biomaterials with broad application, for instance as scaffolds for tissue engineering or as carriers in drug delivery systems (9-11).Despite the keen interest in proteins that undergo s...

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“…Current elastin preservative strategies aim to (a) protect existing elastin against degradation by matrix metalloproteinases (MMP) [6] and elastases [7], (b) replace lost elastin structures (e.g., with synthetic elastomers [8,9], peptide-derived elastomers [10,11], allogeneic elastomers [12]), or (c) regenerate elastin matrices by providing appropriate elastogenic cues (e.g., scaffolds, growth factors). However these approaches have thus far met with only limited success due to non-identification of suitable biochemical cues that can upregulate inherently poor tropoelastin synthesis by adult vascular cells.…”

Section: Introductionmentioning

confidence: 99%

Impact of delivery mode of hyaluronan oligomers on elastogenic responses of adult vascular smooth muscle cells

2007

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Our prior studies demonstrated that exogenous supplements of pure hyaluronan (HA) tetramers (HA4) dramatically upregulate elastin matrix synthesis by adult vascular smooth muscle cells (SMCs). Some studies suggest that exogenous HA likely only transiently contacts and signals cells, and may elicit different cell responses when presented on a substrate (e.g., scaffold surface). To clarify such differences, we used a carbodiimide-based chemistry to tether HA4 onto glass, and compared elastin matrix synthesis by SMCs cultured on these substrates, with those cultured with equivalent amounts of exogenous HA4. Tethered HA4-layers were first characterized for homogeneity, topography, and hydrolytic stability using SEM, XPS, AFM, and FACE. In general, mode of HA4 presentation did not influence its impact on SMC proliferation, or cell synthesis of tropoelastin and matrix elastin, relative to non-HA controls; however, surface-tethered HA4 stimulated SMCs to generate significantly greater amounts of elastin-stabilizing desmosine crosslinks, which partially accounts for the greater resistance to enzymatic breakdown of elastin derived from these cultures. Elastin derived from both sets of cultures contained peptide masses that correspond to the predominant peptides present in rat aortic elastin. SEM and TEM showed that HA4 stimulated fibrillin-mediated elastin matrix deposition, and organization into fibrils. Surfaceimmobilized HA4 was particularly conducive to organization of elastin into aggregating fibrils, and their networking to form closely-woven sheets of elastin fibers, as seen in cardiovascular tissues. The results suggest that incorporation of elastogenic HA4 mers onto cell culture substrates or scaffolds is a better approach than exogenous supplementation for in vitro or in vivo regeneration of architecturally and compositionally faithful-, and more stable mimics of native vascular elastin matrices.

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Recombinant human elastin polypeptides self‐assemble into biomaterials with elastin‐like properties

Cited by 235 publications

References 34 publications

Biomimetic materials for tissue engineering

Biomimetic materials for tissue engineering

Direct observation of structure and dynamics during phase separation of an elastomeric protein

Impact of delivery mode of hyaluronan oligomers on elastogenic responses of adult vascular smooth muscle cells

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