Multiphoton imaging of myogenic differentiation in gelatin-based hydrogels as tissue engineering scaffolds

Min Jeong Kim, Yong Cheol Shin, Jong Ho Lee, Seung Won Jun, Chang Seok Kim, Yunki Lee, Jong Chul Park, Soo Hong Lee, Ki Dong Park, Dong Wook Han

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Background: Hydrogels can serve as three-dimensional (3D) scaffolds for cell culture and be readily injected into the body. Recent advances in the image technology for 3D scaffolds like hydrogels have attracted considerable attention to overcome the drawbacks of ordinary imaging technologies such as optical and fluorescence microscopy. Multiphoton microscopy (MPM) is an effective method based on the excitation of two-photons. In the present study, C2C12 myoblasts differentiated in 3D gelatin hydroxyphenylpropionic acid (GHPA) hydrogels were imaged by using a custom-built multiphoton excitation fluorescence microscopy to compare the difference in the imaging capacity between conventional microscopy and MPM. Results: The physicochemical properties of GHPA hydrogels were characterized by using scanning electron microscopy and Fourier-transform infrared spectroscopy. In addition, the cell viability and proliferation of C2C12 myoblasts cultured in the GHPA hydrogels were analyzed by using Live/Dead Cell and CCK-8 assays, respectively. It was found that C2C12 cells were well grown and normally proliferated in the hydrogels. Furthermore, the hydrogels were shown to be suitable to facilitate the myogenic differentiation of C2C12 cells incubated in differentiation media, which had been corroborated by MPM. It was very hard to get clear images from a fluorescence microscope. Conclusions: Our findings suggest that the gelatin-based hydrogels can be beneficially utilized as 3D scaffolds for skeletal muscle engineering and that MPM can be effectively applied to imaging technology for tissue regeneration.

Original languageEnglish
Article number2
JournalBiomaterials Research
Issue number1
Publication statusPublished - 2016 Jan 1


All Science Journal Classification (ASJC) codes

  • Ceramics and Composites
  • Medicine (miscellaneous)
  • Biomaterials
  • Biomedical Engineering

Cite this

Kim, M. J., Shin, Y. C., Lee, J. H., Jun, S. W., Kim, C. S., Lee, Y., Park, J. C., Lee, S. H., Park, K. D., & Han, D. W. (2016). Multiphoton imaging of myogenic differentiation in gelatin-based hydrogels as tissue engineering scaffolds. Biomaterials Research, 20(1), [2].