Abstract
The extracellular matrix (ECM) is critical to cell attachment, proliferation, and differentiation; therefore, development of bioadaptive ECM-mimetic cell culture scaffolds is an active area of research in tissue engineering. Collagen (rigid nanofiber morphology) and hyaluronan (carboxy groups displayed in a regular manner) are typical ECM components found in vivo. In this work, we used wood-derived crystalline cellulose nanofibers (CNFs) to provide bioadaptive microenvironments for cell culture. Catalytic oxidation of wood CNFs using 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO) afforded crystalline nanofibrous scaffolds with various carboxy contents, ranging from 0.31 to 1.60 mmol g–1, of the TEMPO-oxidized CNFs (TOCNFs). Mouse fibroblast cells were subjected to the hydrophilic TOCNF-coated substrates, which were transparent and exhibited a native cellulose I crystalline structure; good cell proliferation was observed for the TOCNFs with carboxy content of 0.8–1.0 mmol g–1, although intact CNF was bioinert and an excess of carboxylates negatively impacted cell growth. Neither mercerized TOCNFs with a cellulose II structure nor carboxymethylated CNFs with irregular surface carboxy groups contributed to cell proliferation. Therefore, the rigid nanofiber morphology of xeno-free, crystalline TOCNFs with regular alignment of surface carboxy groups would hold the key to providing bioadaptive, ECM-mimetic cellular microenvironments.
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Acknowledgments
The authors appreciate technical assistance from the Center of Advanced Instrumental Analysis, Kyushu University; and the Ultramicroscopy Research Center, Kyushu University. The authors are grateful to Ms. Kana Ichibakase for technical support regarding cell assays.
Funding
This research was funded by the Grant-in-Aid for Scientific Research (KAKENHI) Program (grant numbers JP20K22592 to M.H., JP21K14890 to M.H., JP17H01482 to T.K. and JP18K19233 to T.K.) from the Japan Society for the Promotion of Science, and the Short-term Intensive Research Support Program from the Faculty of Agriculture, Kyushu University (M.H. and T.K.).
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M.H. and T.K. conceived the conception of this work. M.H. conducted all experiments and analytical characterization. T.K. designed research as a project administrator. M.H. and T.K. contributed to writing and reviewing the manuscript. Both authors have read and approved the published version of the manuscript.
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Hatakeyama, M., Kitaoka, T. Surface-Carboxylated Nanocellulose-Based Bioadaptive Scaffolds for Cell Culture. Cellulose 29, 2869–2883 (2022). https://doi.org/10.1007/s10570-021-04154-5
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DOI: https://doi.org/10.1007/s10570-021-04154-5