Towards innervation of bioengineered muscle constructs: Development of a sustained neurotrophic factor delivery and release system

Date publication

août 2022

Journal

Bioprinting

Auteurs

Membres identifiés du Cancéropôle Est :
Dr CLEYMAND Franck, Pr MANO João F.

Tous les auteurs :
Poerio A, Mashanov V, Lai D, Kim M, Ju YM, Kim JH, Lee SJ, Cleymand F, Mano JF, Atala A, Yoo JJ

Résumé

Surgical implantation of biomanufactured skeletal muscle constructs has recently emerged as a promising strategy to treat volumetric muscle defects. However, due to the slow rate of neural regeneration and integration, timely innervation of the implanted constructs with the host peripheral nerves remains an unresolved challenge. This study aims to develop a sustained release neurotrophic factor (NF) delivery system to accelerate peripheral nerve regeneration and innervation of three-dimensional (3D) bioprinted skeletal muscle constructs. Poly (lactic-co-glycolic acid) (PLGA) microspheres were selected as a delivery system for efficient loading and sustained release of two potent NFs: ciliary neurotrophic factor (CNTF) and glial cell line-derived neurotrophic factor (GDNF). We demonstrate that the NFs can be loaded within the PLGA microspheres with a high encapsulation efficiency (75.4% ± 12.6%). The NF-loaded microspheres were incorporated into the fibrinogen-based bioink used to produce biomanufactured skeletal muscle constructs and tested for printability. The microspheres did not change the viscoelastic properties of the bioink, nor did they affect the viability of human muscle progenitor cells. The release kinetic test confirmed that the bioprinted muscle constructs with the NFs-loaded microspheres released the NFs in a sustained manner compared to the bioprinted muscle construct without microspheres. The released NFs maintained their biological activities. In an in vitro neurite outgrowth assay, the NFs released from the PLGA microspheres facilitated the neurite growth over a longer time scale than the NFs directly loaded in the hydrogel. These results demonstrate the feasibility of incorporating the microsphere-based NF delivery system for accelerating neural regeneration in future in vivo applications involving biomanufactured muscle constructs.