Development and evaluation of a removable tissue-engineered muscle with artificial tendons.
Tissue-engineered skeletal muscles were potentially useful as physiological and biochemical in vitro models. Currently, most of the similar models were constructed without tendons. In this study, we aimed to develop a simple, highly versatile tissue-engineered muscle with artificial tendons, and to evaluate the contractile, histological and molecular dynamics during differentiation. C2C12 cells were embedded in a cold type-І collagen gel and placed between two artificial tendons on a silicone sheet. The construct shrank and tightly attached to the artificial tendons with differentiation, finally detaching from the silicone sheet within 1 week of culture onset. We successfully developed a tissue-engineered skeletal muscle with two artificial tendons from C2C12 myoblasts embedded in type-І collagen gel. The isometric twitch contractile force (TCF) significantly increased during differentiation. Time to Peak Tension (TPT) and Half-Relaxation Time (1/2RT) were significantly shortened during differentiation. Myogenic regulatory factors were maximally expressed at 2 weeks, and subsequently decreased at 3 weeks of culture. Histological analysis indicated that myotube formation increased markedly from 2 weeks and well-ordered sarcomere structures were observed on the surface of the 3D engineered muscle at 3 weeks of culture. These results suggested that robust muscle structure occurred by 3 weeks in the tissue-engineered skeletal muscle. Moreover, during the developmental process, the artificial tendons might contribute to well-ordered sarcomere formation. Our results indicated that this simple culture system could be used to evaluate the effects of various pharmacological and mechanical cues on muscle contractility in a variety of research areas.
Division of Human Sciences, Faculty of Engineering, Osaka Institute of Technology, 5-16-1 Ohmiya, Asahi-ku, Osaka 535-8585, Japan. Electronic address: tomohiro.nakamura@oit.ac.jp.
Tomohiro
T
Nakamura
Biomedical Engineering Graduate School of Engineering, Osaka Institute of Technology, 5-16-1 Ohmiya, Asahi-ku, Osaka 535-8585, Japan. Electronic address: d1d13h01@st.oit.ac.jp.
Shunya
S
Takagi
Biomedical Engineering Graduate School of Engineering, Osaka Institute of Technology, 5-16-1 Ohmiya, Asahi-ku, Osaka 535-8585, Japan. Electronic address: t-kamon@kawasumi.jp.
Takafumi
T
Kamon
Biomedical Engineering Graduate School of Engineering, Osaka Institute of Technology, 5-16-1 Ohmiya, Asahi-ku, Osaka 535-8585, Japan. Electronic address: kenichi.yamasaki@fukuda.co.jp.
Ken-Ichi
KI
Yamasaki
Biomedical Engineering Graduate School of Engineering, Osaka Institute of Technology, 5-16-1 Ohmiya, Asahi-ku, Osaka 535-8585, Japan. Electronic address: toshiya.fujisato@oit.ac.jp.
Toshia
T
Fujisato
D000818:Animals; D001011:Aorta / Q000166:cytology; D058045:Artificial Cells / Q000166:cytology* / Q000637:transplantation; D002454:Cell Differentiation; D002460:Cell Line; D003094:Collagen / Q000737:chemistry; D051379:Mice; D008809:Mice, Inbred C3H; D009119:Muscle Contraction; D024510:Muscle Development; D018482:Muscle, Skeletal / Q000166:cytology* / Q000637:transplantation*; D032446:Myoblasts / Q000166:cytology / Q000637:transplantation; D013552:Swine; D013710:Tendons / Q000166:cytology* / Q000637:transplantation; D023822:Tissue Engineering / Q000379:methods*
D003094:Collagen
C2C12; Muscle contractility; Myogenesis; Myogenic regulatory factors; Tissue-engineered muscle