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

Bruno Paiva dos Santos (1) , Bertrand Garbay (2, 3) , Mattia Pasqua (1) , Elsa Chevron (1) , Zoeisha Chinoy (2, 3) , Christophe Cullin (4) , Katell Bathany (4) , Sébastien Lecommandoux (2, 3) , Joelle Amedee (1) , Hugo Oliveira (1) , Elisabeth Garanger (2, 3)

1 BIOTIS - Bioingénierie tissulaire
2 LCPO - Laboratoire de Chimie des Polymères Organiques
3 Team 3 LCPO : Polymer Self-Assembly & Life Sciences
4 CBMN - Chimie et Biologie des Membranes et des Nanoobjets

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.

Mots clés: polyethylene glycol; elastin-like polypeptides; sensory neurons; hydrogels; recombinant expression; thioflavin; inverse transition cycling; transition temperature; tissue-engineering

Domaines: Biotechnologies; Biomatériaux; Matière Molle [cond-mat.soft]

Bruno Paiva dos Santos, Bertrand Garbay, Mattia Pasqua, Elsa Chevron, Zoeisha Chinoy, et al.. Production, purification and characterization of an elastin-like polypeptide containing the Ile-Lys-Val-Ala-Val (IKVAV) peptide for tissue engineering applications. Journal of Biotechnology, 2019, 298, pp.35-44. ⟨10.1016/j.jbiotec.2019.04.010⟩. ⟨hal-02100654⟩