The 2004 Annual Meeting (January 14-20, 2004) of OASYS_NEW

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Contractile Tissue Engineered Skeletal Muscle Incorporating a Vascular Pedicle

Borschel GH, Section of Plastic and Reconstructive Surgery, Department of Surgery, University of Michigan, 2130 Taubman Center, 1500 E. Medical Ctr. Drive, Ann Arbor, MI, USA, Dow DE, Functional Tissue Engineering Laboratory, University of Michigan, Rm 973, 300 N Ingalls Bldg, Ann Arbor, MI, USA, Dennis RG, Departments of Mechanical and Biomedical Engineering, University of Michigan, Ann Arbor, MI, USA, and Brown DL, Section of Plastic Surgery, University of Michigan, 1500 East Medical Center Drive, F7859 Mott, Ann Arbor, MI, USA.

Introduction: Tissue engineered contractile skeletal muscle would be of benefit to patients with disorders such as longstanding facial paralysis or upper extremity muscle loss or dysfunction. Previous investigations found that as three-dimensional muscle constructs grew larger, their function was limited by the lack of a vascular supply. In an effort to expand the potential clinical applicability of engineered muscle, we developed a method a producing contractile engineered skeletal muscle incorporating an axial vascular pedicle in vivo.

Methods: Primary myoblast cultures were generated from adult F344 rats. The cells were trypsinized at 80% confluence and resuspended in a fibrinogen hydrogel. The gel suspension was pipetted into a silicone chamber. The chamber with cell suspension was placed around the common femoral vessels in adult rats. The constructs remained in vivo for three weeks, at which point they were explanted and subjected to isometric force measurements and histologic evaluation (n=8). A control group without cells was also tested (n=6).

Results: The engineered skeletal muscle constructs produced longitudinal contractile force when electrically stimulated, compared with control constructs without myoblasts, which produced no significant force (Wilcoxon rank sum test, p < 0.001). Length-tension, force-voltage, and force-frequency relationships similar to those found in differentiated skeletal muscle were noted. Desmin staining demonstrated that individual myoblasts had undergone fusion and differentiation into multinucleated myotubes. Von Willebrand staining showed that the local environment within the chamber was richly angiogenic, and capillaries had grown into the constructs from the femoral artery and vein.

Conclusions: This new model was capable of generating contractile skeletal muscle tissue. The excitation-contraction and histologic properties of the engineered tissue resembled those of developing skeletal muscle. The incorporation of an axial vascular pedicle provides additional surgical versatility, necessary for the further development of a clinically useful engineered muscle construct.


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