Genetically engineered silk–collagen-like copolymer for biomedical applications: Production, characterization and evaluation of cellular response

Włodarczyk‐Biegun, Małgorzata K.; Werten, Marc W. T.; Wolf, Frits A. de; Beucken, Jeroen J.J.P. van den; Leeuwenburgh, Sander C. G.; Kamperman, Marleen; Stuart, Martien A. Cohen

doi:10.1016/j.actbio.2014.05.006

Cited by 35 publications

(60 citation statements)

References 46 publications

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“…It appears, therefore, that the present D A ::D B pair of heterodimerizing leucine zippers is best used at low concentrations, where the homodimer melting temperatures are low, and one can safely assume that homodimerization does not occur. For example, we have previously designed and produced silk‐like protein polymers that form fibrillar hydrogels (Golinska et al, ; Martens et al, ; Włodarczyk‐Biegun et al, ). By mixing in a very small fraction of fibril‐forming silk‐like protein polymers that additionally have D A and D B domains, we may be able to bundle and/or cross‐link the fibrils in order to modulate the mechanical properties of the gels, as required by many biomedical applications of protein polymer hydrogels (Ehrick et al, ; Seliktar, ; Tan and Marra, ).…”

Section: Discussionmentioning

confidence: 99%

Production in Pichia pastoris of protein‐based polymers with small heterodimer‐forming blocks

Domeradzka

Werten

Vries

et al. 2015

Biotech & Bioengineering

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Some combinations of leucine zipper peptides are capable of forming α-helical heterodimeric coiled coils with very high affinity. These can be used as physical cross-linkers in the design of protein-based polymers that form supramolecular structures, for example hydrogels, upon mixing solutions containing the complementary blocks. Such two-component physical networks are of interest for many applications in biomedicine, pharmaceutics, and diagnostics. This article describes the efficient secretory production of A and B type leucine zipper peptides fused to protein-based polymers in Pichia pastoris. By adjusting the fermentation conditions, we were able to significantly reduce undesirable proteolytic degradation. The formation of A-B heterodimers in mixtures of the purified products was confirmed by size exclusion chromatography. Our results demonstrate that protein-based polymers incorporating functional heterodimer-forming blocks can be produced with P. pastoris in sufficient quantities for use in future supramolecular self-assembly studies and in various applications.

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Section: Discussionmentioning

confidence: 99%

Production in Pichia pastoris of protein‐based polymers with small heterodimer‐forming blocks

Domeradzka

Werten

Vries

et al. 2015

Biotech & Bioengineering

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“…Prior to the studies of SELPs, thermoresponsive synthetic polymers, such as poly(N-isopropylacrylamide) (PNIAAm) [12, 15], and several other classes of recombinant proteins, including elastin-like proteins (ELPs) [11, 53, 54], silk-like proteins (SLPs) [44–46, 55] and collagen-like proteins (CLPs) [56–59], were explored and found to have interesting self-assembling or stimuli responsive properties. PNIPAAm exhibits a lower critical solution temperature (LCST) at about 32°C in water, depending on its concentration and molecular weight.…”

Section: Introductionmentioning

confidence: 99%

Silk-elastin-like protein biomaterials for the controlled delivery of therapeutics

Huang

Rollett

Kaplan

2014

Expert Opinion on Drug Delivery

116

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Introduction Genetically engineered biomaterials are useful for controlled delivery owing to their rational design, tunable structure-function, biocompatibility, degradability and target specificity. Silk-elastin-like proteins (SELPs), a family of genetically engineered recombinant protein polymers, possess these properties. Additionally, given the benefits of combining semicrystalline silk-blocks and elastomeric elastin-blocks, SELPs possess multi-stimuli responsive properties and tenability, thereby, becoming promising candidates for targeted cancer therapeutics delivery and controlled gene release. Areas covered An overview of SELP biomaterials for drug delivery and gene release is provided. Biosynthetic strategies used for SELP production, fundamental physicochemical properties, and self-assembly mechanisms are discussed. The review focuses on sequence-structure-function relationships, stimuli responsive features, and current and potential drug delivery applications. Expert opinion The tunable material properties allow SELPs to be pursued as promising biomaterials for nano-carriers and injectable drug release systems. Current applications of SELPs have focused on thermally-triggered biomaterial formats for the delivery of therapeutics, based on local hyperthermia in tumors or infections. Other prominent controlled release applications of SELPs as injectable hydrogels for gene release have also been pursued. Further biomedical applications that utilize other stimuli to trigger the reversible material responses of SELPs for targeted delivery, including pH, ionic strength, redox, enzymatic stimuli and electric field, are in progress. Exploiting these additional stimuli responsive features will provide a broader range of functional biomaterials for controlled therapeutics release and tissue regeneration.

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“…Its material properties have likewise been manipulated through additions such as carbon nanotubes for hardening20 and poly(lactic-co-glycolic acid) to modify biological degradability21. Efforts have also been made to create silk-based chimeric proteins with sequences sourced from elastin22, collagen232425, and Fn, particularly the RGD (Arg-Gly-Asp) cell binding domain262728, all with the intent to improve native silk bioactivity for biological applications. However, recombinant protein production comes with a substantial methodological barrier while only recapitulating a small degree of function from their parent proteins, such as the adhesive and viability differences of the RGD peptide versus full-length Fn2930.…”

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confidence: 99%

Silk-fibronectin protein alloy fibres support cell adhesion and viability as a high strength, matrix fibre analogue

Jacobsen

Rim

et al. 2017

Sci Rep

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Silk is a natural polymer with broad utility in biomedical applications because it exhibits general biocompatibility and high tensile material properties. While mechanical integrity is important for most biomaterial applications, proper function and integration also requires biomaterial incorporation into complex surrounding tissues for many physiologically relevant processes such as wound healing. In this study, we spin silk fibroin into a protein alloy fibre with whole fibronectin using wet spinning approaches in order to synergize their respective strength and cell interaction capabilities. Results demonstrate that silk fibroin alone is a poor adhesive surface for fibroblasts, endothelial cells, and vascular smooth muscle cells in the absence of serum. However, significantly improved cell attachment is observed to silk-fibronectin alloy fibres without serum present while not compromising the fibres’ mechanical integrity. Additionally, cell viability is improved up to six fold on alloy fibres when serum is present while migration and spreading generally increase as well. These findings demonstrate the utility of composite protein alloys as inexpensive and effective means to create durable, biologically active biomaterials.

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Genetically engineered silk–collagen-like copolymer for biomedical applications: Production, characterization and evaluation of cellular response

Cited by 35 publications

References 46 publications

Production in Pichia pastoris of protein‐based polymers with small heterodimer‐forming blocks

Production in Pichia pastoris of protein‐based polymers with small heterodimer‐forming blocks

Silk-elastin-like protein biomaterials for the controlled delivery of therapeutics

Silk-fibronectin protein alloy fibres support cell adhesion and viability as a high strength, matrix fibre analogue

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