Evaluation of a biomimetic 3D substrate based on the Human Elastin-like Polypeptides (HELPs) model system for elastolytic activity detection

Corich, Lucia; Busetti, Marina; Petix, Vincenzo; Passamonti, Sabina; Bandiera, Antonella

doi:10.1016/j.jbiotec.2017.06.006

Cited by 12 publications

(17 citation statements)

References 34 publications

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“…We set up a method to estimate the stability of HELP deposited on PLLA, by exploiting the susceptibility of HELP to protease-dependent elastolysis (Corich et al, 2017 ). Figure 1C shows that after the wash, cross-linked HELP-PLLA samples retained up to 96% of the HELP protein after the wash step, whereas < 20% of it remained in the non-cross-linked samples.…”

Section: Resultsmentioning

confidence: 99%

“…This strategy is useful to combine the individual performances of the two constituents and future study will clarify the properties of the new hybrid material. The stimuli-responsive nature of the HELP moiety has been already proven and described (Bandiera et al, 2014 ; Corich et al, 2017 ) and it represents an advantage that can be conferred to any new material derived from it. Moreover, HELP-based proteins represent a platform that is readily customizable by molecular fusion of exogenous domains, to confer specific functionality to the final product.…”

Section: Discussionmentioning

confidence: 96%

“…HELP matrix has been shown to possess stimuli-responsive properties, undergoing selective degradation in the presence of elastolytic activity. Exploiting this feature represents an attractive option to realize smart systems that can be employed for therapeutic delivery, tissue engineering, and biosensing (Corich et al, 2017 ).…”

Section: Discussionmentioning

confidence: 99%

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Composite of Elastin-Based Matrix and Electrospun Poly(L-Lactic Acid) Fibers: A Potential Smart Drug Delivery System

Bandiera

Passamonti

Dolci

et al. 2018

Front. Bioeng. Biotechnol.

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Stimuli-responsive hydrogel matrices are inspiring manifold applications in controlled delivery of bioactive compounds. Elastin-derived polypeptides form hydrogel matrices that may release bioactive moieties as a function of local increase of active elastases, as it would occur in several processes like inflammation. In view of the development of a patch for healing wounds, recombinant elastin-based polypeptides were combined with a proteolysis-resistant scaffold, made of electrospun poly-L-lactic acid (PLLA) fibers. The results of this study demonstrated the compatibility of these two components. An efficient procedure to obtain a composite material retaining the main features of each component was established. The release of the elastin moiety was monitored by means of a simple protocol. Our data showed that electrospun PLLA can form a composite with fusion proteins bound to elastin-derived polypeptides. Therefore, our approach allows designing a therapeutic agent delivery platform to realize devices capable of responding and interacting with biological systems at the molecular level.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 96%

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Composite of Elastin-Based Matrix and Electrospun Poly(L-Lactic Acid) Fibers: A Potential Smart Drug Delivery System

Bandiera

Passamonti

Dolci

et al. 2018

Front. Bioeng. Biotechnol.

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“…The synthesis of HELPs was carried out as previously described (Bandiera, ) in the C3037 Escherichia coli strain (New England Biolabs, Ipswich, MA) and the HELP recombinant protein was purified as already detailed (Corich, Busetti, Petix, Passamonti, & Bandiera, ). For the realization of HUG, the synthetic gene of the HELP polypeptide was fused with the 139 amino acids coding sequence of the UnaG bilirubin‐binding protein (accession number BAN57322.1; GenBank), exploiting the unique Dra III site in the expression vector that allows the in‐frame insertion of the polypeptide at the C‐terminus.…”

Section: Methodsmentioning

confidence: 99%

Human elastin‐like polypeptides as a versatile platform for exploitation of ultrasensitive bilirubin detection by UnaG

Bandiera

Corich

Tommasi

et al. 2019

Biotech & Bioengineering

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A new, bifunctional recombinant protein was expressed as the fusion product of human elastin‐like polypeptide (HELP) and the bilirubin‐binding protein UnaG. The engineered product displays both the HELP‐specific property of forming a functional hydrogel matrix and the UnaG‐specific capacity of emitting green fluorescence upon ligand binding. The new fusion protein has been proven to be effective at detecting bilirubin in complex environments with high background noise. A cell culture model of the stress response, consisting of bilirubin released in the cell culture medium, was set up to assess the bilirubin‐sensing properties of the functional matrix obtained by cross‐linking the HELP moiety. Our engineered protein allowed us to monitor cell induction by the release of bilirubin in the culture medium on a nanomolar scale. This study shows that elastin‐like protein fusion represents a versatile platform for the development of novel and commercially viable analytical and biosensing devices.

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“…Several issues have been considered to be essential for the construction of an effective scaffold, namely i) the threedimensional nano-microstructure required, ii) its porosity, iii) its specific mechanical characteristics, iv) its wettability and permeability, v) the need for manipulation and the discomfort caused to the patient, vi) cell-material communication via biochemical signals, and vii) the time of residence before host tissue remodeling [99]. Taking advantage of their recombinant nature, biocompatibility and mechanical features, elastin-like scaffolds have been designed and produced to reproduce some of the properties of the ECM such as architectural topographies, suitable firmness and porosity, promotion of cell adhesion and viability, cell signaling responsiveness, biodegradability and stimuli-responsiveness to changes in their environment, amongst others [100][101][102][103][104]. In this review we describe some different approaches in which elastin has been engineered to obtain customized supports adapted to the part of the body to be treated, such as the skeletal system [105][106][107][108] and vascular system [109][110], and also for the regeneration of soft tissues, with some of the most recent examples being discussed below.…”

Section: Tissue Engineering and Regenerative Medicinementioning

confidence: 99%

Genetically Engineered Elastin-based Biomaterials for Biomedical Applications

Santos

Serrano-Dúcar

González‐Valdivieso

et al. 2020

CMC

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Protein-based polymers are some of the most promising candidates for a new generation of innovative biomaterials as recent advances in genetic-engineering and biotechnological techniques mean that protein-based biomaterials can be designed and constructed with a high degree of complexity and accuracy. Moreover, their sequences, which are derived from structural protein-based modules, can easily be modified to include bioactive motifs that improve their functions and material-host interactions, thereby satisfying fundamental biological requirements. The accuracy with which these advanced polypeptides can be produced, and their versatility, self-assembly behavior, stimuli-responsiveness and biocompatibility, means that they have attracted increasing attention for use in biomedical applications such as cell culture, tissue engineering, protein purification, surface engineering and controlled drug delivery. The biopolymers discussed in this review are elastin-derived protein-based polymers which are biologically inspired and biomimetic materials. This review will also focus on the design, synthesis and characterization of these genetically encoded polymers and their potential utility for controlled drug and gene delivery, as well as in tissue engineering and regenerative medicine.

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Evaluation of a biomimetic 3D substrate based on the Human Elastin-like Polypeptides (HELPs) model system for elastolytic activity detection

Cited by 12 publications

References 34 publications

Composite of Elastin-Based Matrix and Electrospun Poly(L-Lactic Acid) Fibers: A Potential Smart Drug Delivery System

Composite of Elastin-Based Matrix and Electrospun Poly(L-Lactic Acid) Fibers: A Potential Smart Drug Delivery System

Human elastin‐like polypeptides as a versatile platform for exploitation of ultrasensitive bilirubin detection by UnaG

Genetically Engineered Elastin-based Biomaterials for Biomedical Applications

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