Scaffolds For Skeletal Muscle Regeneration Research Articles

Therapeutic applications of biological macromolecules and scaffolds for skeletal muscle regeneration: A review

Skeletal muscle (SM) mass and strength maintenance are important requirements for human well-being. SM regeneration to repair minor injuries depends upon the myogenic activities of muscle satellite (stem) cells. However, losses of regenerative properties following volumetric muscle loss or severe trauma or due to congenital muscular abnormalities are not self-restorable, and thus, these conditions have major healthcare implications and pose clinical challenges. In this context, tissue engineering based on different types of biomaterials and scaffolds provides an encouraging means of structural and functional SM reconstruction. In particular, biomimetic (able to transmit biological signals) and several porous scaffolds are rapidly evolving. Several biological macromolecules/biomaterials (collagen, gelatin, alginate, chitosan, and fibrin etc.) are being widely used for SM regeneration. However, available alternatives for SM regeneration must be redesigned to make them more user-friendly and economically feasible with longer shelf lives. This review aimed to explore the biological aspects of SM regeneration and the roles played by several biological macromolecules and scaffolds in SM regeneration in cases of volumetric muscle loss.

Porous biomaterial scaffolds for skeletal muscle tissue engineering.

Volumetric muscle loss is a traumatic injury which overwhelms the innate repair mechanisms of skeletal muscle and results in significant loss of muscle functionality. Tissue engineering seeks to regenerate these injuries through implantation of biomaterial scaffolds to encourage endogenous tissue formation and to restore mechanical function. Many types of scaffolds are currently being researched for this purpose. Scaffolds are typically made from either natural, synthetic, or conductive polymers, or any combination therein. A major criterion for the use of scaffolds for skeletal muscle is their porosity, which is essential for myoblast infiltration and myofiber ingrowth. In this review, we summarize the various methods of fabricating porous biomaterial scaffolds for skeletal muscle regeneration, as well as the various types of materials used to make these scaffolds. We provide guidelines for the fabrication of scaffolds based on functional requirements of skeletal muscle tissue, and discuss the general state of the field for skeletal muscle tissue engineering.

Silver nanowire loaded poly(ε-caprolactone) nanocomposite fibers as electroactive scaffolds for skeletal muscle regeneration

Volumetric muscle loss (VML) due to trauma and tumor removal operations affects millions of people every year. Although skeletal muscle has a natural repair mechanism, it cannot provide self-healing above a critical level of VML. In this study, nanocomposite aligned fiber scaffolds as support materials were developed for volumetric skeletal muscle regeneration. For this purpose, silver nanowire (Ag NW) loaded poly(ε-caprolactone) (PCL) nanocomposite fiber scaffolds (PCL-Ag NW) were prepared to mimic the aligned electroactive structure of skeletal muscle and provide topographic and conductive environment to modulate cellular behavior and orientation. A computer-aided rotational wet spinning (RWS) system was designed to produce high-yield fiber scaffolds. Nanocomposite fiber bundles with lengths of 50 cm were fabricated via this computer-aided RWS system. The morphological, chemical, thermal properties and biodegradation profiles of PCL and PCL-Ag NW nanocomposite fibers were characterized in detail. The proliferation behavior and morphology of C2C12 mouse myoblasts were investigated on PCL and PCL-Ag NW nanocomposite fibrous scaffolds with and without electrical stimulation. Significantly enhanced cell proliferation was observed on PCL-Ag NW nanocomposite fibers compared to neat PCL fibers with electrical stimulations of 1.5 V, 3 V and without electrical stimulation.

In-vitro effectiveness of poly-β-alanine reinforced poly(3-hydroxybutyrate) fibrous scaffolds for skeletal muscle regeneration

In skeletal muscle tissue engineering, success has not been achieved yet, since the properties of the tissue cannot be fully mimicked. The aim of this study is to investigate the potential use of poly-3-hydroxybutyrate (P3HB)/poly-β-alanine (PBA) fibrous tissue scaffolds with piezoelectric properties for skeletal muscle regeneration. Random and aligned P3HB/PBA (5:1) fibrous matrices were prepared by electrospinning with average diameters of 951 ± 153 nm and 891 ± 247 nm, respectively. X-ray diffraction (XRD) analysis showed that PBA reinforcement and aligned orientation of fibers reduced the crystallinity and brittleness of P3HB matrix. While tensile strength and elastic modulus of random fibrous matrices were determined as 3.9 ± 1.0 MPa and 86.2 ± 10.6 MPa, respectively, in the case of aligned fibers they increased to 8.5 ± 1.8 MPa and 378.2 ± 4.2 MPa, respectively. Aligned matrices exhibited a soft and an elastic behaviour with ~70% elongation in similar to the natural tissue. For the first time, d33 piezoelectric modulus of P3HB/PBA matrices were measured as 5 pC/N and 5.3 pC/N, for random and aligned matrices, respectively. Cell culture studies were performed with C2C12 myoblastic cell line. Both of random and aligned P3HB/PBA fibrous matrices supported attachment and proliferation of myoblasts, but cells cultured on aligned fibers formed regular and thick myofibril structures similar to the native muscle tissue. Reverse transcription polymerase chain reaction (RT-qPCR) analysis indicated that MyoD gene was expressed in the cells cultured on both fiber orientation, however, on the aligned fibers significant increase was determined in Myogenin and Myosin Heavy Chain (MHC) gene expressions, which indicate functional tubular structures. The results of RT-qPCR analysis were also supported with immunohistochemistry for myogenic markers. These in vitro studies have shown that piezoelectric P3HB/PBA aligned fibrous scaffolds can successfully mimic skeletal muscle tissue with its superior chemical, morphological, mechanical, and electroactive properties.

Modeling the mechanics of fibrous-porous scaffolds for skeletal muscle regeneration.

The scaffolds for skeletal muscle regeneration are designed to mimic the structure, stiffness, and strains applied to the muscle under physiologic conditions. The external strains are also used to stimulate myogenesis of the (stem) cells seeded on the scaffold. The time- and location-dependent mechanics inside the scaffold determine the microenvironment for the seeded cells. Here, fibrous-porous cylindrical scaffolds under the action of external cyclic strains are considered. The scaffold mechanics are described as two-phase (poroelastic) where the solid phase is associated with the fibers and the fluid phase is associated with the liquid-containing pores. In response to an applied cyclic strain, pressure oscillates and fluid moves radially toward and away from the axis of the scaffold. We compute the directions and magnitudes of the radial gradients of the poroelastic characteristics (solid-phase displacement, strain, and velocity; fluid-phase pressure and velocity; relative fluid-solid-phase velocity) determined by the boundary conditions and geometry of the scaffold. Several kinds of the external cyclic strain are analyzed and the resulting poroelastic functions are found. The poroelastic characteristics are obtained in closed form which is convenient for further consideration of myogenesis of the seeded cells and ultimately for the design of the scaffolds for skeletal muscle regeneration. Graphical abstract.

3D Graphene Scaffolds for Skeletal Muscle Regeneration: Future Perspectives.

Although skeletal muscle can regenerate after injury, in chronic damages or in traumatic injuries its endogenous self-regeneration is impaired. Consequently, tissue engineering approaches are promising tools for improving skeletal muscle cells proliferation and engraftment. In the last decade, graphene and its derivates are being explored as novel biomaterials for scaffolds production for skeletal muscle repair. This review describes 3D graphene-based materials that are currently used to generate complex structures able not only to guide cell alignment and fusion but also to stimulate muscle contraction thanks to their electrical conductivity. Graphene is an allotrope of carbon that has indeed unique mechanical, electrical and surface properties and has been functionalized to interact with a wide range of synthetic and natural polymers resembling native musculoskeletal tissue. More importantly, graphene can stimulate stem cell differentiation and has been studied for cardiac, neuronal, bone, skin, adipose, and cartilage tissue regeneration. Here we recapitulate recent findings on 3D scaffolds for skeletal muscle repairing and give some hints for future research in multifunctional graphene implants.

Biomimetic Scaffolds in Skeletal Muscle Regeneration.

Skeletal muscle tissue has inherent capacity for regeneration in response to minor injuries. However, in the case of severe trauma, tumor ablations, or in congenital muscle defects, these myopathies can cause irreversible loss of muscle mass and function, a condition referred to as volumetric muscle loss (VML). The natural muscle repair mechanisms are overwhelmed, prompting the search for new muscle regenerative strategies, such as using biomaterials that can provide regenerative signals to either transplanted or host muscle cells. Recent studies involve the use of suitable biomaterials which may be utilized as a template to guide tissue reorganization and ultimately provide optimum micro-environmental conditions to cells. These strategies range from approaches that utilize biomaterials alone to those that combine materials with exogenous growth factors, and ex vivo cultured cells. A number of scaffold materials have been used in the development of grafts to treat VML. In this brief review, we outline the natural skeletal regeneration process, available treatments used in the clinic for muscle injury and promising tissue bioengineering and regenerative approaches for muscle loss treatment.

Optimizing C2C12 myoblast differentiation using polycaprolactone-polypyrrole copolymer scaffolds.

Advancements in tissue engineering and biomaterial development have the potential to provide a scalable solution to the problem of large-volume skeletal muscle defects. Previous research on the development of scaffolds for skeletal muscle regeneration has focused on strategies for increasing conductivity, which has improved satellite cell attachment and differentiation. However, these strategies usually increase scaffold stiffness, which some studies suggest may be detrimental to myoblast development. In this study, the polymers polypyrrole (PPy) and polycaprolactone (PCL) were synthesized together into a copolymer (PPy-PCL) designed to increase scaffold conductivity without significantly influencing stiffness. Different scaffold groups were fabricated via electrospinning, characterized, and assessed for their suitability for myoblast proliferation and differentiation. The groups included an aligned and random iteration of pure PCL, 10% PPy-PCL, 20% PPy-PCL, and 40% PPy-PCL. Only the 40% PPy-PCL group had a measureable conductivity, and the addition of PPy-PCL had no significant effect on the stiffness of the scaffolds. The PPy-PCL copolymer significantly increased the attachment of C2C12 myoblasts as compared to pure PCL scaffolds, but the concentration of PPy-PCL did not significantly alter cell attachment. In addition, scaffolds with PPy-PCL promoted myoblast differentiation to a greater extent than scaffolds made of PCL as measured by fusion index and number of nuclei per myotube. Aligned scaffolds were superior to random scaffolds in almost all measures. These results suggest that conductivity may not be the key factor in improving skeletal muscle scaffolds. Instead, cell attachment and aligned guidance cues may have a greater impact on myoblast differentiation. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 220-231, 2019.

5-Azacytidine-mediated hMSC behavior on electrospun scaffolds for skeletal muscle regeneration.

Incomplete regeneration after trauma or muscular dysfunction is a common problem in muscle replacement therapies. Recent approaches in tissue engineering allow for the replication of skeletal muscle structure and function in vitro and in vivo by molecular therapies and implantable scaffolds which properly address muscle cells toward myotube differentiation and maturation. Here, we investigate the in vitro response of human mesenchymal stem cells (hMSC) on electrospun fibers made of polycaprolactone (PCL) in the presence of 5-azacytidine (5-AZA) to evaluate how fibrous network may influence the therapeutic effect of drug during in vitro myogenesis. Biological studies demonstrate the ability of hMSCs to differentiate in mature myofibers in supplemented (myogenic) and, preferentially, in 5-AZA-enriched culture. PCL electrospun fibers amplify the 5-AZA capability to induce a low proliferation rate in hMSC, thus promoting hMSC differentiation (MTT assay). Qualitative (Azan Mallory stain, immunofluorescence assay, SEM analyses) and quantitative (ELISA test) assays confirm the synergistic contribution of PCL electrospun fibers and 5-AZA on in vitro myotubes formation and maturation. This result is also confirmed by the expression of muscle-specific proteins related to the myogenic mechanisms in the presence of other muscle inductive signals (i.e., oxytocin, Tweak). Hence, we suggest the use of PCL electrospun fibers as interesting preclinical model to explore the effect of drugs and chemotherapeutics administration after damaged muscle resection. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2551-2561, 2017.