Hybrid hydrogel-aligned carbon nanotube scaffolds to enhance cardiac differentiation of embryoid bodies

Samad Ahadian; Shukuyo Yamada; Javier Ramón-Azcón; Mehdi Estili; Xiaobin Liang; Ken Nakajima; Hitoshi Shiku; Ali Khademhosseini; Tomokazu Matsue

doi:10.1016/j.actbio.2015.11.047

Hybrid hydrogel-aligned carbon nanotube scaffolds to enhance cardiac differentiation of embryoid bodies

Acta Biomater. 2016 Feb:31:134-143. doi: 10.1016/j.actbio.2015.11.047. Epub 2015 Nov 24.

Authors

Samad Ahadian¹, Shukuyo Yamada², Javier Ramón-Azcón³, Mehdi Estili⁴, Xiaobin Liang¹, Ken Nakajima¹, Hitoshi Shiku², Ali Khademhosseini⁵, Tomokazu Matsue⁶

Affiliations

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 Nanobiotechnology for Diagnostics (Nb4D) Group, IQAC-CSIC, Jordi Girona 18-26, Barcelona 08034, Spain; CIBER de Bioingenieria, Biomateriales y Nanomedicina (CIBER-BBN), Jordi Girona 18-26, Barcelona 08034, Spain.
⁴ Advanced Ceramics Group, Materials Processing Unit, National Institute for Materials Science (NIMS), Tsukuba 305-0047, Japan.
⁵ WPI-Advanced Institute for Materials Research, Tohoku University, Sendai 980-8577, Japan; Department of Medicine, Center for Biomedical Engineering, Brigham and Women's Hospital, Harvard Medical School, Cambridge, MA 02139, USA; Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, MA 02139, USA; Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02115, USA; College of Animal Bioscience and Technology, Department of Bioindustrial Technologies, Konkuk University, Hwayang-dong, Kwangjin-gu, Seoul 143-701, Republic of Korea; Department of Physics, Faculty of Science, King Abdulaziz University, Jeddah 21569, Saudi Arabia. Electronic address: alik@rics.bwh.harvard.edu.
⁶ WPI-Advanced Institute for Materials Research, Tohoku University, Sendai 980-8577, Japan; Graduate School of Environmental Studies, Tohoku University, Sendai 980-8579, Japan. Electronic address: matsue@bioinfo.che.tohoku.ac.jp.

PMID: 26621696
DOI: 10.1016/j.actbio.2015.11.047

Abstract

Carbon nanotubes (CNTs) were aligned in gelatin methacryloyl (GelMA) hydrogels using dielectrophoresis approach. Mouse embryoid bodies (EBs) were cultured in the microwells fabricated on the aligned CNT-hydrogel scaffolds. The GelMA-dielectrophoretically aligned CNT hydrogels enhanced the cardiac differentiation of the EBs compared with the pure GelMA and GelMA-random CNT hydrogels. This result was confirmed by Troponin-T immunostaining, the expression of cardiac genes (i.e., Tnnt2, Nkx2-5, and Actc1), and beating analysis of the EBs. The effect on EB properties was significantly enhanced by applying an electrical pulse stimulation (frequency, 1Hz; voltage, 3V; duration, 10ms) to the EBs for two continuous days. Taken together, the fabricated hybrid hydrogel-aligned CNT scaffolds with tunable mechanical and electrical characteristics offer an efficient and controllable platform for electrically induced differentiation and stimulation of stem cells for potential tissue regeneration and cell therapy applications.

Statement of significance: Dielectrophoresis approach was used to rapidly align carbon nanotubes (CNTs) in gelatin methacryloyl (GelMA) hydrogels resulting in hybrid GelMA-CNT hydrogels with tunable and anisotropic electrical and mechanical properties. The GelMA-aligned CNT hydrogels may be used to apply accurate and controllable electrical pulses to cell and tissue constructs and thereby regulating their behavior and function. In this work, it was demonstrated that the GelMA hydrogels containing the aligned CNTs had superior performance in cardiac differentiation of stem cells upon applying electrical stimulation in contrast with control gels. Due to broad use of electrical stimulation in tissue engineering and stem cell differentiation, it is envisioned that the GelMA-aligned CNT hydrogels would find wide applications in tissue regeneration and stem cell therapy.

Keywords: Carbon nanotubes; Cardiac differentiation; Electrical stimulation; Embryoid body; Hydrogel.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Anisotropy
Biocompatible Materials / chemistry
Cell Differentiation
Cell Line
Cell Survival
DNA, Complementary / metabolism
Electrodes
Embryoid Bodies / cytology*
Gelatin / chemistry
Heart / growth & development*
Hydrogels / chemistry*
Mice
Microscopy, Atomic Force
Nanotubes, Carbon / chemistry*
Polymers / chemistry
Regeneration
Stem Cells / cytology*
Stress, Mechanical
Tissue Engineering / methods
Tissue Scaffolds / chemistry*
Troponin T / chemistry

Substances

Biocompatible Materials
DNA, Complementary
Hydrogels
Nanotubes, Carbon
Polymers
Troponin T
Gelatin