Fabrication and Characterization of Electrospun Decellularized Muscle-Derived Scaffolds

Abstract

Although skeletal muscle has a high potential for self-repair, volumetric muscle loss can result in impairment beyond the endogenous regenerative capacity. There is a clinical need to improve on current clinical treatments that fail to fully restore the structure and function of lost muscle. Decellularized extracellular matrix (dECM) scaffolds have been an attractive platform for regenerating skeletal muscle, as dECM contains many biochemical cues that aid in cell adhesion, proliferation, and differentiation. However, there is limited capacity to tune physicochemical properties in current dECM technologies to improve outcome. In this study, we aim to create a novel, high-throughput technique to fabricate dECM scaffolds with tunable physicochemical properties while retaining proregenerative matrix components. We demonstrate a successful decellularization protocol that effectively removes DNA. We also identified key steps for the successful production of electrospun muscle dECM without the use of a carrier polymer; electrospinning allows for rapid scaffold fabrication with high control over material properties, which can be optimized to mimic native muscle. To this end, fiber orientation and degree of crosslinking of these dECM scaffolds were modulated and the corollary effects on fiber swelling, mechanical properties, and degradation kinetics were investigated. Beyond application in skeletal muscle, the versatility of this technology has the potential to serve as a foundation for dECM scaffold fabrication in a variety of tissue engineering applications.

Advances in biomaterials for skeletal muscle engineering and obstacles still to overcome

“…However, restoration of muscle structure and function were not supported within the short, 4-week study. To further leverage the biochemical cues provided by skeletal muscle dECM with the tunable fiber diameter and alignment afforded through electrospinning, our laboratory developed a method of fabricating electrospun scaffolds completely derived from skeletal muscle dECM with tunable physicochemical properties [ 92 ]. Unlike other natural polymers, the electrospun dECM system demonstrated enhanced versatility in its innate physiologically relevant mechanical properties and degradation kinetics in the absence of a crosslinking agent.…”

Natural hydrogels R&D process: technical and regulatory aspects for industrial implementation

“…Kin and co-workers studied the wound healing process using an electrospinning scaffold based on decellularized cardiac tissue and they demonstrated the material’s ability not only to reduce scarring by rapidly replacing collagen type III with the stronger collagen type I, but also the material angiogenesis promotion with a minimum inflammatory response [ 48 ]. Other decellularized tissues are also used to create an electrospinning scaffold [ 49 ], for instance, muscle tissue [ 50 ] but, all of them take advantage of the decellularized material’s ability to induce cell attachment, proliferation, migration, differentiation, and maturation, even of stem cells [ 51 ]. The reason is the preservation of natural molecules present into the native tissue, such as growth factors, proteoglycans, bioactive cryptic peptides and natural proteins like collagen and fibronectin, which promote biological activities as cell growth, function, differentiation, angiogenesis, antimicrobial effects, and chemotactic effects [ 52 – 54 ].…”

Smart ECM-Based Electrospun Biomaterials for Skeletal Muscle Regeneration

“…This latter result suggests that dECM proteins provide the cues for attachment and growth of satellite cells onto the nanofibers, while the presence of PCL assures structural integrity and elasticity to the scaffold. A study developed a method to fabricate electrospun scaffolds from the decellularized skeletal muscle without the need for a carrier polymer is noteworthy [ 30 ]. The resulting scaffolds showed tunable physicochemical properties, including fiber alignment, while important extracellular matrix components for regeneration such as GAGs, were preserved.…”

“…To overcome these obstacles, some authors are developing ad hoc decellurization protocols for specific tissues and the optimization of subsequent dECM process methods with the aim of avoiding the loss of some important bioactive components of the native ECM [ 29 , 30 ].…”