Generation of a Functioning and Self-Renewing Diaphragmatic Muscle Construct

Abstract

Surgical repair of large muscular defects requires the use of autologous graft transfer or prosthetic material. Naturally derived matrices are biocompatible materials obtained by tissue decellularization and are commonly used in clinical practice. Despite promising applications described in the literature, the use of acellular matrices to repair large defects has been only partially successful, highlighting the need for more efficient constructs. Scaffold recellularization by means of tissue engineering may improve not only the structure of the matrix, but also its ability to functionally interact with the host. The development of such a complex construct is challenging, due to the complexity of the native organ architecture and the difficulties in recreating the cellular niche with both proliferative and differentiating potential during growth or after damage. In this study, we tested a mouse decellularized diaphragmatic extracellular matrix (ECM) previously described by our group, for the generation of a cellular skeletal muscle construct with functional features. The decellularized matrix was stored using different conditions to mimic the off‐the‐shelf clinical need. Pediatric human muscle precursors were seeded into the decellularized scaffold, demonstrating proliferation and differentiation capability, giving rise to a functioning three‐dimensional skeletal muscle structure. Furthermore, we exposed the engineered construct to cardiotoxin injury and demonstrated its ability to activate a regenerative response in vitro promoting cell self‐renewal and a positive ECM remodeling. Functional reconstruction of an engineered skeletal muscle with maintenance of a stem cell pool makes this a promising tool toward future clinical applications in diaphragmatic regeneration. Stem Cells Translational Medicine 2019;8:858&869

Porcine Decellularized Diaphragm Hydrogel: A New Option for Skeletal Muscle Malformations

“…Nowadays, this type of scaffold represents the best compromise between physiologic environment and synthetic materials when used as a substitute in SKM defects. Building upon previous findings in our research group on murine diaphragmatic dECM [24,36], we moved to produce a porcine diaphragmatic dECM to adopt a smart scaled-up approach and potentially manufacture different scaffold types. We adapted the detergent-enzymatic decellularization process used for murine samples to porcine muscle, setting up a reproducible technique to obtain a completely acellular scaffold that resembles native diaphragmatic architecture and protein composition.…”

Extracellular matrix: an important regulator of cell functions and skeletal muscle development

“…In recent years, researchers have developed dECM active materials for clinical treatment using bioengineering techniques. Trevisan et al constructed mouse decellularized diaphragm ECM, which can promote the activation, proliferation and differentiation of skeletal muscle progenitor cells to form a powerful three-dimensional skeletal muscle structure, providing a promising tool for clinical application of diaphragm regeneration in the future [ 159 ]. Lee et al used skeletal muscle-derived dECM and IGF1 to develop a decellularized muscle-specific scaffold system, which can better promote cell proliferation and differentiation, thus supporting in situ regeneration of muscle tissue [ 160 ].…”

Extracellular Matrix-Derived Hydrogels as Biomaterial for Different Skeletal Muscle Tissue Replacements

“…However, the best cell source to reproduce SKM in vitro is still to be found. Several works demonstrate that a mixed population composed of different-if not all-SKM cell components are necessary to generate in vitro a complete and mature muscle substitute [45,50].…”