Biomimetic glycosaminoglycan-based scaffolds improve skeletal muscle regeneration in a Murine volumetric muscle loss model

Naagarajan Narayanan; Zhihao Jia; Kun Ho Kim; Liangju Kuang; Paul Lengemann; Gabrielle Shafer; Victor Bernal-Crespo; Shihuan Kuang; Meng Deng

doi:10.1016/j.bioactmat.2020.10.012

Biomimetic glycosaminoglycan-based scaffolds improve skeletal muscle regeneration in a Murine volumetric muscle loss model

Bioact Mater. 2020 Oct 29;6(4):1201-1213. doi: 10.1016/j.bioactmat.2020.10.012. eCollection 2021 Apr.

Authors

Naagarajan Narayanan^{1

2}, Zhihao Jia³, Kun Ho Kim³, Liangju Kuang^{1

2}, Paul Lengemann^{1

2}, Gabrielle Shafer⁴, Victor Bernal-Crespo⁴, Shihuan Kuang³, Meng Deng^{1

2

5

6}

Affiliations

¹ Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, IN, 47906, United States.
² Bindley Bioscience Center, Purdue University, West Lafayette, IN, 47906, United States.
³ Department of Animal Science, Purdue University, West Lafayette, IN, 47906, United States.
⁴ Center for Comparative Translational Research, Purdue University, West Lafayette, IN, 47906, United States.
⁵ School of Materials Engineering, Purdue University, West Lafayette, IN, 47906, United States.
⁶ Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, 47906, United States.

Abstract

Volumetric muscle loss (VML) injuries characterized by critical loss of skeletal muscle tissues result in severe functional impairment. Current treatments involving use of muscle grafts are limited by tissue availability and donor site morbidity. In this study, we designed and synthesized an implantable glycosaminoglycan-based hydrogel system consisting of thiolated hyaluronic acid (HA) and thiolated chondroitin sulfate (CS) cross-linked with poly(ethylene glycol) diacrylate to promote skeletal muscle regeneration of VML injuries in mice. The HA-CS hydrogels were optimized with suitable biophysical properties by fine-tuning degree of thiol group substitution to support C2C12 myoblast proliferation, myogenic differentiation and expression of myogenic markers MyoD, MyoG and MYH8. Furthermore, in vivo studies using a murine quadriceps VML model demonstrated that the HA-CS hydrogels supported integration of implants with the surrounding host tissue and facilitated migration of Pax7⁺ satellite cells, de novo myofiber formation, angiogenesis, and innervation with minimized scar tissue formation during 4-week implantation. The hydrogel-treated and autograft-treated mice showed similar functional improvements in treadmill performance as early as 1-week post-implantation compared to the untreated groups. Taken together, our results demonstrate the promise of HA-CS hydrogels as regenerative engineering matrices to accelerate healing of skeletal muscle injuries.

Keywords: AChR, Acetyl choline receptors; CS, Chondroitin Sulfate; Chondroitin sulfate; ECM, Extracellular matrix; EDC, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide; GAG, Glycosaminoglycan; HA, Hyaluronic acid; Hyaluronic acid; Hydrogels; MES, 2-(N-morpholino) ethanesulfonic acid; MHC, Myosin heavy chain; Myoblasts; NHS, N-hydroxysuccinimide; PEGDA, Poly(ethylene glycol) diacrylate; Skeletal muscle tissue engineering; VML, Volumetric muscle loss; Volumetric muscle loss; eMHC, embryonic myosin heavy chain.

Abstract

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