Development of Salt Leached Silk Fibroin Scaffold using Direct Dissolution Techniques for Cartilage Tissue Engineering

Untung Ari Wibowo, Hermawan Judawisastra, Anggraini Barlian, Nayla M. Alfarafisa, Karina F Moegni, Melinda Remelia


Autologous transplantations, the gold standard, did not meet sufficient health tissue coverage area for cartilage damage treatments. The field of tissue engineering offers a promising alternative to fulfill this limitation by growing patient own cells on biomaterials through tissue culture, reconstructed into new cartilage tissue, and the implanted to the injury area. To support tissue regeneration, biocompatible, biodegradable, and high strength silk fibroin (SF) was proposed in this study as scaffold materials. In this research, direct dissolution in CaCl2/formic acid, a faster and simpler process than traditional dissolution techniques, combined with salt leaching technique. SF contents on the scaffold were varied from 2 w/v% to 12 w/v% and NaCl size as porogen was fixed in diameter of 250±58 µm. Evaluation of the SF scaffold’s morphology, hydrophilicity, biodegradability, and biocompatibility were conducted. The results showed porous silk fibroin scaffold had been successfully developed. The SF scaffolds have pore size 261-293 µm with highly interconnected pores. FTIR and XRD analysis of the scaffolds showed the characteristics of silk fibroin, which reveals the α-helix amorphous and β-sheet crystalline structure and comparable to the silk fibers. The scaffold showed good hydrophilicity and high water uptakes, which essential properties for cell survival. The scaffold degraded under Protease XIV, indicate biodegradable properties. Observation of cell attachment confirms the scaffold has good biocompatibility to adipose-derived stem cells and are suitable to be used in cartilage tissue engineering.


direct dissolution; porous scaffold; salt leaching; silk fibroin; tissue engineering.

Full Text:



Y. Liu, G. Zhou, and Y. Cao, “Recent Progress in Cartilage Tissue Engineering—Our Experience and Future Directions,†Engineering, vol. 3, no. 1. pp. 28–35, 2017.

J. Lanza, R., Langer, R., Vacanti, Principle of Tissue Engineering, vol. Third Edit. 2007.

F. J. O’Brien, “Biomaterials & scaffolds for tissue engineering,†Mater. Today, vol. 14, no. 3, pp. 88–95, 2011.

B. Kundu, R. Rajkhowa, S. C. Kundu, and X. Wang, “Silk fibroin biomaterials for tissue regenerations,†Adv. Drug Deliv. Rev., vol. 65, no. 4, pp. 457–470, 2013.

E. A. Makris, A. H. Gomoll, K. N. Malizos, J. C. Hu, and K. A. Athanasiou, “Repair and tissue engineering techniques for articular cartilage,†Nat. Rev. Rheumatol., vol. 11, no. 1, pp. 21–34, 2014.

R. Nazarov, H.-J. Jin, and D. L. Kaplan, “Porous 3-D Scaffolds from Regenerated Silk Fibroin,†Biomacromolecules, vol. 5, no. 3, pp. 718–726, May 2004.

F. Zhang, X. You, H. Dou, Z. Liu, B. Zuo, and X. Zhang, “Facile fabrication of robust silk nanofibril films via direct dissolution of silk in CaCl2-formic acid solution,†ACS Appl. Mater. Interfaces, vol. 7, no. 5, pp. 3352–3361, 2015.

F. Zhang et al., “Regeneration of high-quality silk fibroin fiber by wet spinning from CaCl2-formic acid solvent,†Acta Biomater., vol. 12, no. 1, pp. 139–145, 2015.

H. J. Park et al., “Fabrication of 3D porous silk scaffolds by particulate (salt/sucrose) leaching for bone tissue reconstruction,†Int. J. Biol. Macromol., vol. 78, no. Supplement C, pp. 215–223, 2015.

S. Mohanty et al., “Fabrication of scalable tissue engineering scaffolds with dual-pore microarchitecture by combining 3D printing and particle leaching,†Mater. Sci. Eng. C, vol. 61, no. Supplement C, pp. 180–189, 2016.

K. Kanimozhi, S. K. Basha, and V. S. Kumari, “Fabrication of chitosan based hybrid porous scaffolds by salt leaching for soft tissue engineering,†Surfaces and Interfaces, vol. 1–3, no. Supplement C, pp. 7–12, 2016.

F. Zhou et al., “Silk fibroin-chondroitin sulfate scaffold with immuno-inhibition property for articular cartilage repair,†Acta Biomater., vol. 63, no. Supplement C, pp. 64–75, 2017.

U. J. Kim, J. Park, H. Joo Kim, M. Wada, and D. L. Kaplan, “Three-dimensional aqueous-derived biomaterial scaffolds from silk fibroin,†Biomaterials, vol. 26, no. 15, pp. 2775–2785, 2005.

M. Li, M. Ogiso, and N. Minoura, “Enzymatic degradation behavior of porous silk fibroin sheets,†Biomaterials, vol. 24, no. 2, pp. 357–365, 2003.

G.-I. Im, J.-Y. Ko, and J. H. Lee, “Chondrogenesis of Adipose Stem Cells in a Porous Polymer Scaffold: Influence of the Pore Size,†Cell Transplant., vol. 21, no. 11, pp. 2397–2405, 2012.

X. Zhang, C. Cao, X. Ma, and Y. Li, “Optimization of macroporous 3-D silk fibroin scaffolds by salt-leaching procedure in organic solvent-free conditions,†J. Mater. Sci. Mater. Med., vol. 23, no. 2, pp. 315–324, 2012.

X. Hu, D. Kaplan, and P. Cebe, “Determining beta-sheet crystallinity in fibrous proteins by thermal analysis and infrared spectroscopy,†Macromolecules, vol. 39, no. 18, pp. 6161–6170, 2006.

S. Ling, Z. Qi, D. P. Knight, Z. Shao, and X. Chen, “FTIR imaging, a useful method for studying the compatibility of silk fibroin-based polymer blends,†Polym. Chem., vol. 4, no. 21, p. 5401, 2013.

X. J. Lian, S. Wang, and H. S. Zhu, “Surface properties and cytocompatibillity of silk fibroin films cast from aqueous solutions in different concentrations,†Front. Mater. Sci. China, vol. 4, no. 1, pp. 57–63, 2010.

S. Hofmann et al., “Silk fibroin as an organic polymer for controlled drug delivery,†J. Control. Release, vol. 111, no. 1–2, pp. 219–227, 2006.

N. Minoura, S. Aiba, M. Higuchi, Y. Gotoh, M. Tsukada, and Y. Imai, “Attachment and growth of fibroblast cells on silk fibroin.,†Biochemical and biophysical research communications, vol. 208, no. 2. pp. 511–516, 1995.

J. L. McGrath, “Cell Spreading: The Power to Simplify,†Current Biology, vol. 17, no. 10. 2007.

X. Xie et al., “Comparative evaluation of MSCs from bone marrow and adipose tissue seeded in PRP-derived scaffold for cartilage regeneration,†Biomaterials, vol. 33, no. 29, pp. 7008–7018, 2012.

H. J. Kim, S.-H. Park, J. Durham, J. M. Gimble, D. L. Kaplan, and J. L. Dragoo, “In vitro chondrogenic differentiation of human adipose-derived stem cells with silk scaffolds.,†J. Tissue Eng., vol. 3, no. 1, p. 2041731412466405, 2012.

S.-N. Jung et al., “In vivo cartilage formation using chondrogenic-differentiated human adipose-derived mesenchymal stem cells mixed with fibrin glue.,†J. Craniofac. Surg., vol. 21, no. 2, pp. 468–72, 2010.

K. Dzobo et al., “Fibroblast-derived extracellular matrix induces chondrogenic differentiation in human adipose-derived mesenchymal stromal/stem cells in vitro,†Int. J. Mol. Sci., vol. 17, no. 8, 2016.



  • There are currently no refbacks.

Published by INSIGHT - Indonesian Society for Knowledge and Human Development