3D cellulose nanofiber scaffold with homogeneous cell population and long-term proliferation
Tumor-originated and undefined extracellular matrices (ECMs) such as Matrigel™ have been widely used in three-dimensional (3D) cell and tissue culture, but their use is unacceptable in clinical cell therapies. In this study, we proposed a 3D cellulose nanofiber (CNF) hydrogel that has great potential as a defined tissue-engineering scaffold, especially for osteoblast culture. The CNF hydrogel showed attractive features as a cell scaffold material. It exhibited a ~ 1.4-fold higher diffusion coefficient (~ 2.98 × 10−7 cm2/s) of macromolecules such as bovine serum albumin than does Matrigel™ (< 2.2 × 10−7 cm2/s) due to the former’s higher porosity (> 95%) and pore size (~ 310.8 μm). Most pre-osteoblast cells that are encapsulated in the CNF hydrogel were immediately locked without sinking by instant hydrogen bond cross-linking between CNFs, whereas cells encapsulated in Matrigel™ sank to the bottom of the scaffold due to the slow sol–gel transition (> 20 min). The elastic modulus of the cell-encapsulated CNF hydrogel could be reinforced by further calcium-mediated cross-linking without cytotoxicity. As a result, the pre-osteoblast cells in the CNF hydrogels were homogeneously distributed in the 3D structure, proliferated for 3 weeks, and successfully differentiated. Overall, CNFs showed that it has potential to be used in tissue engineering as a defined ECM component.
KeywordsCellulose nanofibers Extracellular matrices Matrigel Shear-dependent viscosity 3D hydrogel scaffolds
We would like to acknowledge the financial support from the Grant the Marine Biotechnology program (Marine BioMaterials Research Center) funded by the Ministry of Oceans and Fisheries of Korea (D11013214H480000110). This work was also supported by the National Research Foundation of Korea (NRF) funded by the Ministry of Science, ICT and Future Planning (NRF-2016M1A5A1027592 and NRF-2017R1A2B3006354). DXO acknowledges KRICT SI-1809.
Compliance with ethical standards
Conflicts of interest
All authors declare that there is no conflict of interest.
- Chatterjee K et al (2010) The effect of 3D hydrogel scaffold modulus on osteoblast differentiation and mineralization revealed by combinatorial screening. Biomaterials 31(19):5051–5062. https://doi.org/10.1016/j.biomaterials.2010.03.024 CrossRefPubMedPubMedCentralGoogle Scholar
- Evangelista MB et al (2007) Upregulation of bone cell differentiation through immobilization within a synthetic extracellular matrix. Biomaterials 28(25):3644–3655. https://doi.org/10.1016/j.biomaterials.2007.04.028 CrossRefPubMedGoogle Scholar
- Frith JE et al (2013) An injectable hydrogel incorporating mesenchymal precursor cells and pentosan polysulphate for intervertebral disc regeneration. Biomaterials 34(37):9430–9440. https://doi.org/10.1016/j.biomaterials.2013.08.072 CrossRefPubMedGoogle Scholar
- Jing J, Fournier A, Szarpak-Jankowska A, Block MR, Auzély-Velty R (2015) Type, density, and presentation of grafted adhesion peptides on polysaccharide-based hydrogels control preosteoblast behavior and differentiation. Biomacromol 16(3):715–722. https://doi.org/10.1021/bm501613u CrossRefGoogle Scholar
- Karageorgiou V, Kaplan D (2005) Porosity of 3D biomaterial scaffolds and osteogenesis. Biomaterials 26(27):5474–5491. https://doi.org/10.1016/j.biomaterials.2005.02.002 CrossRefPubMedGoogle Scholar
- Malinen MM, Kanninen LK, Corlu A, Isoniemi HM, Lou Y-R, Yliperttula ML, Urtti AO (2014) Differentiation of liver progenitor cell line to functional organotypic cultures in 3D nanofibrillar cellulose and hyaluronan-gelatin hydrogels. Biomaterials 35(19):5110–5121. https://doi.org/10.1016/j.biomaterials.2014.03.020 CrossRefPubMedGoogle Scholar
- Nguyen HL, Jo YK, Cha M, Cha YJ, Yoon DK, Sanandiya ND, Prajatelistia E, Oh DX, Hwang DS (2016) Mussel-inspired anisotropic nanocellulose and silver nanoparticle composite with improved mechanical properties, electrical conductivity and antibacterial activity. Polymers 8(3):102. https://doi.org/10.3390/polym8030102 CrossRefGoogle Scholar
- Nguyen HL, Hanif J, Park SA, Choi BG, Tran TH, Hwang DS, Park JY, Hwang SY, Oh DX (2018) Sustainable boron nitride nanosheet-reinforced cellulose nanofiber composite film with oxygen barrier without the cost of color and cytotoxicity. Polymers 10(5):501. https://doi.org/10.3390/polym10050501 CrossRefGoogle Scholar
- O’Brien FJ, Harley B, Yannas IV, Gibson LJ (2005) The effect of pore size on cell adhesion in collagen-GAG scaffolds. Biomaterials 26(4):433–441. https://doi.org/10.1016/j.biomaterials.2004.02.052 CrossRefPubMedGoogle Scholar
- Oh SH, Park IK, Kim JM, Lee JH (2007) In vitro and in vivo characteristics of PCL scaffolds with pore size gradient fabricated by a centrifugation method. Biomaterials 28(9):1664–1671. https://doi.org/10.1016/j.biomaterials.2006.11.024 CrossRefPubMedGoogle Scholar
- Story BJ, Wagner WR, Gaisser DM, Cook SD, Rust-Dawicki AM (1998) In vivo performance of a modified CSTi dental implant coating. Int J Oral Maxillofacc Implants 13(6):749–757Google Scholar
- Weaver VM, Petersen OW, Wang F, Larabell C, Briand P, Damsky C, Bissell MJ (1997) Reversion of the malignant phenotype of human breast cells in three-dimensional culture and in vivo by integrin blocking antibodies. J Cell Biol 137(1):231–245. https://doi.org/10.1083/jcb CrossRefPubMedPubMedCentralGoogle Scholar