Primary Progenitor Muscle Cells for Regenerative Medicine: Standardization of Therapeutic Protocols and Optimized In Vivo Murine Model For Volumetric Muscle Loss

Abstract Engineering of biomimetic tissue implants provides an opportunity for repairing volumetric muscle loss (VML), beyond a tissue’s innate repair capacity. Here, we present thick, suturable, and pre-vascularized 3D muscle implants containing human induced pluripotent stem cell-derived myogenic precursor cells (hiPSC-MPCs), which can differentiate into skeletal muscle cells while maintaining a self-renewing pool. The formation of contractile myotubes and millimeter-long fibers from hiPSC-MPCs is achieved in chemically, mechanically, and structurally tailored extracellular matrix-based hydrogels, which can serve as scaffolds to ultimately organize the linear fusion of myoblasts. Embedded multi-material bioprinting is used to deposit complex patterns of perfusable vasculatures and aligned hiPSC-MPC channels within an endomysium-like supporting gel to recapitulate muscle architectural integrity in a facile yet highly rapid manner. Moreover, we demonstrate successful graft-host integration and de novo muscle formation upon in vivo implantation of pre-vascularized constructs within a VML model. This work pioneers the engineering of large pre-vascularized hiPSC-derived muscle tissues toward next generation VML regenerative therapies.

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Effects of the tetrahedral framework nucleic acids on the skeletal muscle regeneration in vitro and in vivo

Materials Chemistry Frontiers ◽

10.1039/d0qm00329h ◽

2020 ◽

Vol 4 (9) ◽

pp. 2731-2743

Author(s):

Yang Gao ◽

Tianxu Zhang ◽

Junyao Zhu ◽

Dexuan Xiao ◽

Mei Zhang ◽

...

Keyword(s):

Tissue Regeneration ◽

Muscle Regeneration ◽

Skeletal Muscle Regeneration ◽

Muscle Loss ◽

Degenerative Diseases ◽

Tetrahedral Framework ◽

Volumetric Muscle Loss ◽

Regeneration In Vitro

The challenges associated with muscle degenerative diseases and volumetric muscle loss (VML) emphasizes the prospects of muscle tissue regeneration.

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In Silico and In Vivo Studies Detect Functional Repair Mechanisms in a Volumetric Muscle Loss Injury

Tissue Engineering Part A ◽

10.1089/ten.tea.2018.0280 ◽

2019 ◽

Vol 25 (17-18) ◽

pp. 1272-1288 ◽

Cited By ~ 10

Author(s):

Juliana A. Passipieri ◽

Xiao Hu ◽

Ellen Mintz ◽

Jack Dienes ◽

Hannah B. Baker ◽

...

Keyword(s):

In Silico ◽

In Vivo Studies ◽

Muscle Loss ◽

Volumetric Muscle Loss ◽

Repair Mechanisms

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Matured Myofibers in Bioprinted Constructs with In Vivo Vascularization and Innervation

Gels ◽

10.3390/gels7040171 ◽

2021 ◽

Vol 7 (4) ◽

pp. 171

Author(s):

Catherine G. Y. Ngan ◽

Anita Quigley ◽

Richard J. Williams ◽

Cathal D. O’Connell ◽

Romane Blanchard ◽

...

Keyword(s):

Neuromuscular Diseases ◽

Structural Development ◽

Muscle Loss ◽

Test Drug ◽

Scaffold Material ◽

Volumetric Muscle Loss ◽

Primary Mouse ◽

First Time

For decades, the study of tissue-engineered skeletal muscle has been driven by a clinical need to treat neuromuscular diseases and volumetric muscle loss. The in vitro fabrication of muscle offers the opportunity to test drug-and cell-based therapies, to study disease processes, and to perhaps, one day, serve as a muscle graft for reconstructive surgery. This study developed a biofabrication technique to engineer muscle for research and clinical applications. A bioprinting protocol was established to deliver primary mouse myoblasts in a gelatin methacryloyl (GelMA) bioink, which was implanted in an in vivo chamber in a nude rat model. For the first time, this work demonstrated the phenomenon of myoblast migration through the bioprinted GelMA scaffold with cells spontaneously forming fibers on the surface of the material. This enabled advanced maturation and facilitated the connection between incoming vessels and nerve axons in vivo without the hindrance of a scaffold material. Immunohistochemistry revealed the hallmarks of tissue maturity with sarcomeric striations and peripherally placed nuclei in the organized bundles of muscle fibers. Such engineered muscle autografts could, with further structural development, eventually be used for surgical reconstructive purposes while the methodology presented here specifically has wide applications for in vitro and in vivo neuromuscular function and disease modelling.

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Regenerative and Rehabilitative Medicine: A Necessary Synergy for Functional Recovery from Volumetric Muscle Loss Injury

Cells Tissues Organs ◽

10.1159/000444673 ◽

2016 ◽

Vol 202 (3-4) ◽

pp. 237-249 ◽

Cited By ~ 15

Author(s):

Sarah M. Greising ◽

Christopher L. Dearth ◽

Benjamin T. Corona

Keyword(s):

Skeletal Muscle ◽

Regenerative Medicine ◽

Muscle Loss ◽

Research Fields ◽

Current State ◽

Volumetric Muscle Loss ◽

Us Government ◽

The Usa ◽

Joint Range ◽

Regenerative Therapies

Volumetric muscle loss (VML) is a complex and heterogeneous problem due to significant traumatic or surgical loss of skeletal muscle tissue. The consequences of VML are substantial functional deficits in joint range of motion and skeletal muscle strength, resulting in life-long dysfunction and disability. Traditional physical medicine and rehabilitation paradigms do not address the magnitude of force loss due to VML and related musculoskeletal comorbidities. Recent advancements in regenerative medicine have set forth encouraging and emerging therapeutic options for VML injuries. There is significant potential that combined rehabilitative and regenerative therapies can restore limb and muscle function following VML injury in a synergistic manner. This review presents the current state of the VML field, spanning clinical and preclinical literature, with particular focus on rehabilitation and regenerative medicine in addition to their synergy. Moving forward, multidisciplinary collaboration between clinical and research fields is encouraged in order to continue to improve the treatment of VML injuries and specifically address the encompassing physiology, pathology, and specific needs of this patient population. This is a work of the US Government and is not subject to copyright protection in the USA. Foreign copyrights may apply. Published by S. Karger AG, Basel

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Adipose-derived Stem/Stromal Cells on Electrospun Fibrin Microfiber Bundles Enable Moderate Muscle Reconstruction in a Volumetric Muscle Loss Model

Cell Transplantation ◽

10.1177/0963689718805370 ◽

2018 ◽

Vol 27 (11) ◽

pp. 1644-1656 ◽

Cited By ~ 13

Author(s):

Jordana Gilbert-Honick ◽

Brian Ginn ◽

Yuanfan Zhang ◽

Sara Salehi ◽

Kathryn R. Wagner ◽

...

Keyword(s):

Treatment Options ◽

Donor Site ◽

Donor Site Morbidity ◽

Current Treatment ◽

Muscle Loss ◽

Post Transplantation ◽

Volumetric Muscle Loss ◽

Volume Retention

Current treatment options for volumetric muscle loss (VML) are limited due to donor site morbidity, lack of donor tissue, and insufficient functional recovery. Tissue-engineered skeletal muscle grafts offer the potential to significantly improve functional outcomes. In this study, we assessed the potential pro-myogenic effects of human adipose-derived stem cells (ASCs) seeded onto electrospun uniaxially aligned fibrin hydrogel microfiber bundles. Although both uninduced and 5-azacytidine-induced ASCs exhibited alignment, elongation, and diffuse muscle marker expression when grown on microfiber bundles for 2 months in vitro, both groups failed to fully recapitulate myotube characteristics. To assess the muscle regeneration potential of ASCs in vivo, ASC-seeded fibrin microfiber bundles were implanted in a robust murine VML defect model. Minimal fibrosis was observed surrounding implanted acellular hydrogel fibers at 2 and 4 weeks, and fibers seeded with ASCs exhibited up to 4 times higher volume retention than acellular fibers. We observed increased numbers of cells positive for the regenerating muscle marker embryonic myosin and the mature muscle marker myosin heavy chain in ASC-seeded fibers compared with acellular fibers at 1 and 3 months post-transplantation. Regenerating muscle cells were closely associated with ASC-derived cells and in some cases had potentially fused with them. These findings demonstrate that despite failing to undergo myogenesis in vitro, ASCs combined with electrospun fibrin microfibers moderately increased muscle reconstruction in vivo compared with acellular fibers following a severe VML defect.

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