A 3D bioprinted complex structure for engineering the muscle–tendon unit

Three-dimensional integrated organ printing (IOP) technology seeks to fabricate tissue constructs that can mimic the structural and functional properties of native tissues. This technology is particularly useful for complex tissues such as those in the musculoskeletal system, which possess regional differences in cell types and mechanical properties. Here, we present the use of our IOP system for the processing and deposition of four different components for the fabrication of a single integrated muscle-tendon unit (MTU) construct. Thermoplastic polyurethane (PU) was co-printed with C2C12 cell-laden hydrogel-based bioink for elasticity and muscle development on one side, while poly(ϵ-caprolactone) (PCL) was co-printed with NIH/3T3 cell-laden hydrogel-based bioink for stiffness and tendon development on the other. The final construct was elastic on the PU-C2C12 muscle side (E = 0.39 ± 0.05 MPa), stiff on the PCL-NIH/3T3 tendon side (E = 46.67 ± 2.67 MPa) and intermediate in the interface region (E = 1.03 ± 0.14 MPa). These constructs exhibited >80% cell viability at 1 and 7 d after printing, as well as initial tissue development and differentiation. This study demonstrates the versatility of the IOP system to create integrated tissue constructs with region-specific biological and mechanical characteristics for MTU engineering.

[1]  D. Goldspink,et al.  Muscle growth in response to mechanical stimuli. , 1995, The American journal of physiology.

[2]  George J Christ,et al.  The influence of electrospun aligned poly(epsilon-caprolactone)/collagen nanofiber meshes on the formation of self-aligned skeletal muscle myotubes. , 2008, Biomaterials.

[3]  Anthony Atala,et al.  3D bioprinting of tissues and organs , 2014, Nature Biotechnology.

[4]  Y. Imai,et al.  Transplantation of a tissue-engineered pulmonary artery. , 2001, The New England journal of medicine.

[5]  J. Malda,et al.  Biofabrication of multi-material anatomically shaped tissue constructs , 2013, Biofabrication.

[6]  Kyle W. Binder,et al.  Regenerative medicine as applied to general surgery. , 2012, Annals of surgery.

[7]  J. Wang.  Mechanobiology of tendon. , 2006, Journal of biomechanics.

[8]  Milica Radisic,et al.  Biomaterial based cardiac tissue engineering and its applications , 2015, Biomedical materials.

[9]  S W Herring,et al.  Paralysis and growth of the musculoskeletal system in the embryonic chick , 1990, Journal of morphology.

[10]  Joel D Stitzel,et al.  Co-electrospun dual scaffolding system with potential for muscle-tendon junction tissue engineering. , 2011, Biomaterials.

[11]  W. Liu,et al.  Repair of tendon defect with dermal fibroblast engineered tendon in a porcine model. , 2006, Tissue engineering.

[12]  Sara Mantero,et al.  Clinical transplantation of a tissue-engineered airway , 2008, The Lancet.

[13]  Helen H. Lu,et al.  Tissue Engineering Strategies for the Regeneration of Orthopedic Interfaces , 2010, Annals of Biomedical Engineering.

[14]  H. Vandenburgh,et al.  Mechanical stimulation improves tissue-engineered human skeletal muscle. , 2002, American journal of physiology. Cell physiology.

[15]  Anthony Atala,et al.  Engineering Complex Tissues , 2012, Science Translational Medicine.

[16]  L. Galatz,et al.  The role of mechanobiology in tendon healing. , 2012, Journal of shoulder and elbow surgery.

[17]  D. Cho,et al.  Bioprinting of a mechanically enhanced three-dimensional dual cell-laden construct for osteochondral tissue engineering using a multi-head tissue/organ building system , 2012 .

[18]  H. Vandenburgh.  Dynamic mechanical orientation of skeletal myofibers in vitro. , 1982, Developmental biology.

[19]  Karl Grosh,et al.  Engineering of functional tendon. , 2004, Tissue engineering.

[20]  Taiji Sohmura,et al.  Three-Dimensional Cell and Tissue Patterning in a Strained Fibrin Gel System , 2007, PloS one.

[21]  M. Järvinen,et al.  Mechanical loading regulates the expression of tenascin-C in the myotendinous junction and tendon but does not induce de novo synthesis in the skeletal muscle , 2003, Journal of Cell Science.

[22]  Masood A. Machingal,et al.  Further development of a tissue engineered muscle repair construct in vitro for enhanced functional recovery following implantation in vivo in a murine model of volumetric muscle loss injury. , 2012, Tissue engineering. Part A.

[23]  Albert J Banes,et al.  Novel system for engineering bioartificial tendons and application of mechanical load. , 2003, Tissue engineering.

[24]  P. Bártolo,et al.  Additive manufacturing of tissues and organs , 2012 .

[25]  James J. Yoo,et al.  Bioprinting technology and its applications. , 2014, European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery.

[26]  Ellen M Arruda,et al.  Structure and functional evaluation of tendon-skeletal muscle constructs engineered in vitro. , 2006, Tissue engineering.

[27]  W. Dhert,et al.  Three-dimensional fiber deposition of cell-laden, viable, patterned constructs for bone tissue printing. , 2008, Tissue engineering. Part A.

[28]  Dietmar W Hutmacher,et al.  Scaffold-based tissue engineering: rationale for computer-aided design and solid free-form fabrication systems. , 2004, Trends in biotechnology.

[29]  Johnna S Temenoff,et al.  Engineering orthopedic tissue interfaces. , 2009, Tissue engineering. Part B, Reviews.

[30]  N. Turner,et al.  Functional skeletal muscle formation with a biologic scaffold. , 2010, Biomaterials.

[31]  D. Kohane,et al.  Engineering vascularized skeletal muscle tissue , 2005, Nature Biotechnology.

[32]  James J. Yoo,et al.  Tissue-engineered autologous bladders for patients needing cystoplasty , 2006, The Lancet.

[33]  Yilin Cao,et al.  Engineering human neo-tendon tissue in vitro with human dermal fibroblasts under static mechanical strain. , 2009, Biomaterials.

[34]  J. Tidball,et al.  Mechanical loading regulates expression of talin and its mRNA, which are concentrated at myotendinous junctions. , 1998, American journal of physiology. Cell physiology.

[35]  Joel D Stitzel,et al.  Cyclic mechanical preconditioning improves engineered muscle contraction. , 2008, Tissue engineering. Part A.

[36]  Thomas L. Smith,et al.  A naturally derived, cytocompatible, and architecturally optimized scaffold for tendon and ligament regeneration. , 2007, Biomaterials.

[37]  Hod Lipson,et al.  Direct Freeform Fabrication of Seeded Hydrogels in Arbitrary Geometries , 2022 .