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3D-Printed Graphene Material Scaffolds for Neuromuscular Tissue Engineering

Sumanas W. Jordan, MD, PhD; Adam E. Jakus, PhD; Philip Lewis, BS; Ramille N. Shah, PhD
The Ohio State University

Presenter: Sumanas Jordan

Dr. Jordan conceived of the study and performed all procedures. She interpreted the results and drafted the abstract.

Director Name: Albert Chao

Author Category: Fellow Plastic Surgery
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction

Though vascularization has been a long-recognized challenge in tissue engineering, the task of neurotization has been understated. Functional tissue engineering requires neurotization for applications including facial reanimation, intrinsic hand muscle reconstruction, cardiac rehabilitation, and urologic tone. Electroactive biomaterials, including graphene, have been shown by us and others to differentiate stem cells into neural, glial, and muscle cell types even in the absence of stimulation. This is the first study to evaluate graphene in a neurotized in vivo environment.

Three graphene-based material scaffolds were 3D-printed (8-layers, 600 micron struts, 0-90 degree offset): (i) 3D-printed graphene (3DG, 60%vol graphene/40%vol polylactide-co-glycolide (PLG)), (ii) 3D-printed graphene-decellularized muscle ECM (mECM) blend (3DG-mECM, 30%vol graphene/30%vol mECM particles/40%vol PLG), and (iii) 3DG infused with mECM hydrogel (3DG-mgel). Six-millimeter constructs were implanted in rat medial thigh muscle defects with and without femoral nerve coaptation for 6 weeks. Explants were analyzed by histology and immunohistochemistry.

Material properties of 3DG-mECM blends were consistent with previous 3DG blends. Vascularization was observed between the struts of all constructs. Increased graphene dispersion between struts was observed in the neurotized group compared to internal controls. Myoblasts (Pax7+) were observed replacing the struts of 3DG-mECM constructs. Neurofilament-200 (NF200) staining was seen in all groups. Macrophage (CD68+) amount and distribution did not differ significantly.

Neurotization altered the degradation pattern of the constructs. 3D-printed scaffolds containing decellularized skeletal muscle induced myogenesis without exogenous growth factors or cells. Material ink composition may be tailored for muscle tissue engineering.

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