Dielectrophoretical fabrication of hybrid carbon nanotubes-hydrogel biomaterial for muscle tissue engineering applicatio
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Dielectrophoretical fabrication of hybrid carbon nanotubes-hydrogel biomaterial for muscle tissue engineering applications Javier Ramón-Azcón1, Samad Ahadian1, Raquel Obregon2, Hitoshi Shiku2, Ali Khademhosseini1,4, Tomokazu Matsue1,2 1 WPI-Advanced Institute for Materials Research, Tohoku University, Sendai 980-8577, Japan 2 Graduate School of Environmental Studies, Tohoku University, Sendai 980-8579, Japan 3 Department of Medicine, Center for Biomedical Engineering, Brigham and Women’s Hospital, Harvard Medical School, Cambridge, Massachusetts 02139, USA; Harvard–MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, Massachusetts 02115, USA; Department of Maxillofacial Biomedical Engineering and Institute of Oral Biology, School of Dentistry, Kyung Hee University, Seoul 130-701, Republic of Korea. ABSTRACT Dielectrophoresis (DEP) approach was employed to achieve highly aligned multi-walled carbon nanotubes (MWCNTs) within the gelatin methacrylate (GelMA) hydrogels in a facile, rapid, inexpensive, and reproducible manner. This approach enabled us to make different CNTs alignments (e.g., vertical or horizontal alignments) within the GelMA hydrogel using different electrode designs or configurations. Anisotropically aligned GelMA-CNTs hydrogels showed considerably higher conductivity compared to randomly distributed CNTs dispersed in the GelMA hydrogel and the pristine and non-conductive GelMA hydrogel. Adding 0.3 mg/mL CNTs to the GelMA hydrogel led to a slight increase in the mechanical properties of the GelMA and made it to behave as a viscoelastic material. Therefore, it can be used as a suitable scaffold for soft tissues, such as skeletal muscle tissue. 3D microarrays of skeletal muscle myofibers were then fabricated based on the GelMA and GelMA-CNTs hydrogels and they were characterized in terms of gene expressions related to the muscle cell differentiation and contraction. Owing to high electrical conductivity of aligned GelMA-CNTs hydrogels, the engineered muscle tissues cultivated on these materials demonstrated superior maturation and functionality particularly after applying the electrical stimulation (voltage 8 V, frequency 1 Hz, and duration 10 ms for 2 days) compared to the corresponding tissues obtained on the pristine GelMA and randomly distributed CNTs within the GelMA hydrogel. INTRODUCTION In vitro generation of tissues in principle involves the proliferation and differentiation of cells within a scaffold mimicking the formation of native tissues within the extracellular matrix (ECM). Hydrogels have been considered as ideal materials to mimic the ECM due to high water content and biodegradability [1,2]. However, they generally have weak mechanical properties and low conductivity, which have limited their ability to engineer soft and electroactive tissues, such as skeletal muscle, cardiac, and neural tissues. Therefore, it is desirable to control and
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