Successful engineering of biomimetic skeletal muscle tissues could allow creation of accurate models of muscle physiology and disease and aid treatment of various muscle disorders. This project is based on our recently developed methods to utilize adult rat myogenic cells for engineering of 3D skeletal muscle tissues with structural and functional properties comparable to those of native muscle. Specifically, we have established conditions for robust in vitro expansion of adult rat myogenic cells and have successfully utilized them to engineer skeletal muscle tissues with contractile capacity 10- 100 fold higher than previously reported. Importantly, in a comprehensive set of preliminary studies we for the first time show that self-regenerative capacity of adult-derived engineered muscle in vitro and survival in vivo can be significantly enhanced by a 3-D co-culture of skeletal muscle progenitors with non-polarized macrophages derived from bone marrow. We propose to build on these exciting results and systematically explore the use of 3D muscle-macrophage co-culture system to create highly contractile and regenerative muscle tissues with the capacity for rapid vascular and neuronal integration and successful repair of skeletal muscle injury in vivo. We will study: (1) the cellular and molecular mechanisms of macrophage mediated self-repair of tissue-engineered muscle in vitro, (2) the combined effects of macrophages, vascular cells, and biophysical cues on the ability of in vitro formed pre- vascularized engineered muscle to undergo rapid blood perfusion and functional maturation in vivo, and (3) the roles of macrophage supplementation and synaptogenic stimulation in vitro upon the ability of muscle-macrophage implants to functionally integrate with and repair damaged skeletal muscle in vivo. Successful completion of the proposed studies will establish foundation for the future applications of tissue engineering methodologies to human muscle repair. Furthermore, our novel strategy to utilize immune system cells as pro-regenerative adjuvants inside tissue-engineered implants may find broad applications in the field of regenerative medicine.

Public Health Relevance

Transplantation of engineered skeletal muscles able to self-regenerate and functionally integrate in vivo may lead to efficient repair of damaged or diseased muscle. In this proposal we will develop regeneration and integration-ready functional skeletal muscle tissues and validate their therapeutic potential in rodent models of muscle injury and loss.

Agency
National Institute of Health (NIH)
Institute
National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS)
Type
Research Project (R01)
Project #
5R01AR070543-03
Application #
9532574
Study Section
Musculoskeletal Tissue Engineering Study Section (MTE)
Program Officer
Wang, Fei
Project Start
2016-09-01
Project End
2021-08-31
Budget Start
2018-09-01
Budget End
2019-08-31
Support Year
3
Fiscal Year
2018
Total Cost
Indirect Cost
Name
Duke University
Department
Biomedical Engineering
Type
Biomed Engr/Col Engr/Engr Sta
DUNS #
044387793
City
Durham
State
NC
Country
United States
Zip Code
27705
Juhas, Mark; Abutaleb, Nadia; Wang, Jason T et al. (2018) Incorporation of macrophages into engineered skeletal muscle enables enhanced muscle regeneration. Nat Biomed Eng 2:942-954
Rao, Lingjun; Qian, Ying; Khodabukus, Alastair et al. (2018) Engineering human pluripotent stem cells into a functional skeletal muscle tissue. Nat Commun 9:126
Khodabukus, Alastair; Madden, Lauran; Prabhu, Neel K et al. (2018) Electrical stimulation increases hypertrophy and metabolic flux in tissue-engineered human skeletal muscle. Biomaterials :
Khodabukus, Alastair; Prabhu, Neel; Wang, Jason et al. (2018) In Vitro Tissue-Engineered Skeletal Muscle Models for Studying Muscle Physiology and Disease. Adv Healthc Mater 7:e1701498
Polstein, Lauren R; Juhas, Mark; Hanna, Gabi et al. (2017) An Engineered Optogenetic Switch for Spatiotemporal Control of Gene Expression, Cell Differentiation, and Tissue Morphogenesis. ACS Synth Biol 6:2003-2013