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J Biotechnol. 2019 Jun 10;298:35-44. doi: 10.1016/j.jbiotec.2019.04.010. Epub 2019 Apr 10.

Production, purification and characterization of an elastin-like polypeptide containing the Ile-Lys-Val-Ala-Val (IKVAV) peptide for tissue engineering applications.

Author information

1
Université de Bordeaux, 146 rue Léo Saignat, Bordeaux, 33076, France; INSERM, Bioingénierie Tissulaire (U1026, BioTis), Bordeaux, France.
2
Université de Bordeaux/Bordeaux INP, ENSCBP, 16 avenue Pey-Berland, Pessac, 33607, France; CNRS, Laboratoire de Chimie des Polymères Organiques (UMR5629, LCPO), Pessac, France. Electronic address: bertrand.garbay@bordeaux-inp.fr.
3
Université de Bordeaux/Bordeaux INP, ENSCBP, 16 avenue Pey-Berland, Pessac, 33607, France; CNRS, Laboratoire de Chimie des Polymères Organiques (UMR5629, LCPO), Pessac, France.
4
Université de Bordeaux/Bordeaux INP, allée Geoffroy Saint Hilaire, Pessac, 33600, France; CNRS, Chimie et Biologie des Membranes et des Nano-objets (UMR5248, CBMN), Pessac, France.

Abstract

Elastin-like polypeptides (ELPs) are biocompatible-engineered polypeptides, with promising interest in tissue engineering due to their intrinsic biological and physical properties, and their ease of production. The IKVAV (Ile-Lys-Val-Ala-Val) laminin-1 sequence has been shown to sustain neuron attachment and growth. In this study, the IKVAV adhesion sequence, or a scrambled VKAIV sequence, were incorporated by genetic engineering in the structure of an ELP, expressed in Escherichia coli and purified. The transition temperatures of the ELP-IKVAV and ELP-VKAIV were determined to be 23 °C. Although the phase transition was fully reversible for ELP-VKAIV, we observed an irreversible aggregation for ELP-IKVAV. The corresponding aggregates shared some characteristics with amyloid-like polypeptides. The two ELPs were then reacted with functionalized polyethylene glycol (PEG) to form hydrogels. These hydrogels were characterized for rheological properties, tested with cultures of rat primary sensory neurons, and implanted subcutaneously in mice for 4 weeks. Sensory neurons cultured on high IKVAV concentration hydrogels (20%) formed longer neurite than those of neurons grown on hydrogels containing the scrambled IKVAV sequence. Finally, in vivo evaluation showed the absence of detectable inflammation. In conclusion, this functionalized ELP-IKVAV biomaterial shows interesting properties for tissue engineering requiring neurotization.

KEYWORDS:

Elastin-like polypeptides; Hydrogels; Recombinant expression; Sensory neurons; Tissue-engineering

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