Advertisement

The Maturation of Tissue-Engineered Skeletal Muscle Units Following 28-Day Ectopic Implantation in a Rat

  • Brittany L. Rodriguez
  • Shelby E. Florida
  • Keith W. VanDusen
  • Brian C. Syverud
  • Lisa M. LarkinEmail author
Article
  • 52 Downloads

Abstract

Volumetric muscle loss (VML) is a loss of skeletal muscle that results in a sustained impairment of function and is often accompanied by physical deformity. To address the need for more innovative repair options, our laboratory has developed scaffold-free, multiphasic tissue-engineered skeletal muscle units (SMUs) to treat VML injuries. In our previous work, using the concept of the “body as a bioreactor”, we have shown that implantation promotes the maturation of our SMUs beyond what is possible in vitro. Thus, in this study, we sought to better understand the effect of implantation on the maturation of our SMUs, including the effects of implantation on SMU force production and cellular remodeling. We used an ectopic implantation so that we could more easily dissect the implanted tissues post-recovery and measure the force contribution of the SMU alone and compare it to pre-implantation values. This study also aimed to scale up the size of our SMUs to enable the replacement of larger volumes of muscle in our future VML studies. Overall, implantation resulted in extensive maturation of the SMUs, as characterized by an increase in force production, substantial integration with native tissue, innervation, vascularization, and the development of structural organization similar to native tissue.

Lay Summary

To address the need for more innovative repair options for severe muscle injuries, our laboratory has developed a lab-grown, living muscle tissue for implantation. In our previous work, we have shown that implantation of our engineered tissue promotes its maturation beyond what is possible in the lab. Thus, in this study, we sought to scale-up the size of our engineered muscle and to better understand the effect of implantation on the maturation of our engineered muscle, including the effects on the force production. Overall, implantation resulted in extensive maturation, including an increase in force production, innervation, and vascularization.

In an effort to make this technology more clinically relevant, future work will involve the development and implantation of larger quantities of engineered tissue to replace a larger percentage of the lost muscle. We believe this will help restore muscle function to that of pre-injury levels.

Keywords

Satellite cell Scaffold-free Tissue engineering Skeletal muscle Implantation 

Notes

Funding

The authors would like to acknowledge the support of the NIH/NIAMS R01 grant: 1R01AR067744-01, as well as NIH Research Supplement to Promote Diversity in Health Related Research: 3R01AR067744-02W1.

Compliance with Ethical Standard

All animal care and animal surgery procedures were in accordance with The Guide for Care and Use of Laboratory Animals [6] and the protocol was approved by the University Committee for the Use and Care of Animals.

Conflict of Interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Zouraq F, et al., Skeletal Muscle Regeneration for Clinical Application. Regenerative Medicine and Tissue Engineering, 2013: p. 679–712.Google Scholar
  2. 2.
    Mertens JP, Sugg KB, Lee JD, Larkin LM. Engineering muscle constructs for the creation of functional engineered musculoskeletal tissue. Regen Med. 2014;9(1):89–100.CrossRefGoogle Scholar
  3. 3.
    Grogan BF, Hsu JR, Consortium STR. Volumetric muscle loss. J Am Acad Orthop Surg. 2011;19(Suppl 1):S35–7.CrossRefGoogle Scholar
  4. 4.
    Tedesco FS, Dellavalle A, Diaz-Manera J, Messina G, Cossu G. Repairing skeletal muscle: regenerative potential of skeletal muscle stem cells. J Clin Invest. 2010;120(1):11–9.CrossRefGoogle Scholar
  5. 5.
    VanDusen KW, Syverud BC, Williams ML, Lee JD, Larkin LM. Engineered skeletal muscle units for repair of volumetric muscle loss in the tibialis anterior muscle of a rat. Tissue Eng Part A. 2014;20(21–22):2920–30.CrossRefGoogle Scholar
  6. 6.
    National Research Council. Guide for the care and use of laboratory animals. Washington D.C: The National Academies Press; 2011.Google Scholar
  7. 7.
    Syverud BC, VanDusen KW, Larkin LM. Effects of dexamethasone on satellite cells and tissue engineered skeletal muscle units. Tissue Eng Part A. 2016;22(5–6):480–9.CrossRefGoogle Scholar
  8. 8.
    Williams ML, Kostrominova TY, Arruda EM, Larkin LM. Effect of implantation on engineered skeletal muscle constructs. J Tissue Eng Regen Med. 2013;7(6):434–42.CrossRefGoogle Scholar
  9. 9.
    Larkin LM, Calve S, Kostrominova TY, Arruda EM. Structure and functional evaluation of tendon-skeletal muscle constructs engineered in vitro. Tissue Eng. 2006;12(11):3149–58.CrossRefGoogle Scholar
  10. 10.
    Smietana MJ, Syed-Picard FN, Ma J, Kostrominova T, Arruda EM, Larkin LM. The effect of implantation on scaffoldless three-dimensional engineered bone constructs. In Vitro Cell Dev Biol Anim. 2009;45(9):512–22.CrossRefGoogle Scholar
  11. 11.
    Mahalingam VD, Behbahani-Nejad N, Ronan EA, Olsen TJ, Smietana MJ, Wojtys EM, et al. Fresh versus frozen engineered bone-ligament-bone grafts for sheep anterior cruciate ligament repair. Tissue Eng Part C Methods. 2015;21(6):548–56.CrossRefGoogle Scholar
  12. 12.
    Florida SE, et al., In vivo structural and cellular remodeling of engineered bone-ligament-bone constructs used for anterior cruciate ligament reconstruction in sheep. Connect Tissue Res, 2016: p. 1–13.Google Scholar
  13. 13.
    Ma J, Goble K, Smietana M, Kostrominova T, Larkin L, Arruda EM. Morphological and functional characteristics of three-dimensional engineered bone-ligament-bone constructs following implantation. J Biomech Eng. 2009;131(10):101017.CrossRefGoogle Scholar
  14. 14.
    Larkin LM, et al. Effect of age and neurovascular grafting on the mechanical function of medial gastrocnemius muscles of Fischer 344 rats. J Gerontol A Biol Sci Med Sci. 1998;53(4):B252–8.CrossRefGoogle Scholar
  15. 15.
    Dennis RG, Kosnik PE. Excitability and isometric contractile properties of mammalian skeletal muscle constructs engineered in vitro. In Vitro Cell Dev Biol Anim. 2000;36(5):327–35.CrossRefGoogle Scholar
  16. 16.
    Service, R.F. Tissue engineering. Technique uses body as 'bioreactor' to grow new bone. Science. 2005;309(5735):683.CrossRefGoogle Scholar
  17. 17.
    Stevens MM, Marini RP, Schaefer D, Aronson J, Langer R, Shastri VP. In vivo engineering of organs: the bone bioreactor. Proc Natl Acad Sci U S A. 2005;102(32):11450–5.CrossRefGoogle Scholar
  18. 18.
    Holt GE, Halpern JL, Dovan TT, Hamming D, Schwartz HS. Evolution of an in vivo bioreactor. J Orthop Res. 2005;23(4):916–23.CrossRefGoogle Scholar
  19. 19.
    Warnke PH, Springer ING, Wiltfang J, Acil Y, Eufinger H, Wehmöller M, et al. Growth and transplantation of a custom vascularised bone graft in a man. Lancet. 2004;364(9436):766–70.CrossRefGoogle Scholar
  20. 20.
    Warnke PH, Wiltfang J, Springer I, Acil Y, Bolte H, Kosmahl M, et al. Man as living bioreactor: fate of an exogenously prepared customized tissue-engineered mandible. Biomaterials. 2006;27(17):3163–7.CrossRefGoogle Scholar
  21. 21.
    Jana T, Khabbaz E, Bush CM, Prosser JD, Birchall MA, Nichols CA, et al. The body as a living bioreactor: a feasibility study of pedicle flaps for tracheal transplantation. Eur Arch Otorhinolaryngol. 2013;270(1):181–6.CrossRefGoogle Scholar
  22. 22.
    Laurance J. British boy receives trachea transplant built with his own stem cells. BMJ. 2010;340:c1633.CrossRefGoogle Scholar
  23. 23.
    Dvir T, Kedem A, Ruvinov E, Levy O, Freeman I, Landa N, et al. Prevascularization of cardiac patch on the omentum improves its therapeutic outcome. Proc Natl Acad Sci U S A. 2009;106(35):14990–5.CrossRefGoogle Scholar
  24. 24.
    Syverud BC, Gumucio JP, Rodriguez BL, Wroblewski OM, Florida SE, Mendias CL, et al. A transgenic tdTomato rat for cell migration and tissue engineering applications. Tissue Eng Part C Methods. 2018;24(5):263–71.CrossRefGoogle Scholar

Copyright information

© The Regenerative Engineering Society 2018

Authors and Affiliations

  • Brittany L. Rodriguez
    • 1
  • Shelby E. Florida
    • 2
  • Keith W. VanDusen
    • 2
  • Brian C. Syverud
    • 1
  • Lisa M. Larkin
    • 1
    • 2
    Email author
  1. 1.Biomedical EngineeringUniversity of MichiganAnn ArborUSA
  2. 2.Molecular and Integrative PhysiologyUniversity of MichiganAnn ArborUSA

Personalised recommendations