Iranian Journal of Biomedical Engineering (IJBME)
نشریه علمی مهندسی پزشکی زیستی

Analysis of the Effect of Shear Stress on the Differentiation Response of Mesenchymal Stem Cells in Multi-Layered Nanocomposite Scaffold for Bone-Cartilage Tissue Engineering using Computational Methods

Document Type : Full Research Paper

Authors

Sara Zadegan 1
Bahman Vahidi 2
Nooshin Haghighipour 3

1 M.Sc., Biomedical Engineering Department, Faculty of New Sciences and Technologies (FNST), University of Tehran, Tehran, Iran

2 Associate Professor, Biomedical Engineering Department, Faculty of New Sciences and Technologies (FNST), University of Tehran, Tehran, Iran

3 Professor, Pasteur Institute of Iran (IPI), Tehran, Iran

https://doi.org/10.22041/ijbme.2023.1989428.1828
Abstract
Repairing osteochondral defects (OCD) remains a formidable challenge due to the high complexity of native osteochondral tissue and the limited self-repair capability of cartilage. In this regard, the development of osteochondral tissue engineering with scaffolds seeded with stem cells along with mechanical stimulation has been considered by the researchers as a new proposed technique for the repair of this tissue. In this study, at first we fabricated an integrated and biomimetic trilayered Silk Fibroin (SF) scaffold containing SF nano fibers in each layer. Then fluid wall shear stress in different areas of the scaffold was predicted in dynamic cell culture condition under the inlet velocity of 0.4 ml/min in a perfusion bioreactor using finite elements and fluid-structure interactions methods. Finally, using the simulation results, osteogenesis and chondrogenesis of rabbit adipose derived stem cells (RADSCs) were analyzed. The results showed that this novel osteochondral graft has a seamlessly integrated layer structure and a high degree of pore interconnectivity. The average size of the pores in the bone layer, middle layer, and cartilage were 76, 152, and 102 microns, respectively. In addition, this biomimetic scaffold presented compressive moduli of 0.4 MPa and uitimate tensile strength of 10 MPa in the wet state. Also, based on the simulation analyses, the shear stress distribution is more uniform if the bone layer is exposed to the fluid inlet path which facilitates bone differentiation. Good adhesion and infiltration of cells were observed after 14 days dynamic culture. The results of expression analysis of differentiated genes in bone and cartilage layer containing RADSc after 21 days of culture under static and dynamic conditions showed that perfusion flow significantly upregulated the expression of bone and cartilage genes in the respective layers and downregulated the hypertrophy gene expression in intermediate layer of scaffold.

Keywords

Perfusion Bioreactor
Osteochondral Tissue
Multi Layered Scaffold
Computational Fluid Dynamics
Stem Cells
Silk Fibroin

Subjects

  1. Upmeier, B. Brüggenjürgen, A. Weiler, C. Flamme, H. Laprell, and S. Willich, "Follow-up costs up to 5 years after conventional treatments in patients with cartilage lesions of the knee," Knee Surgery, Sports Traumatology, Arthroscopy, vol. 15, pp. 249-257, 2007.
  2. J. Yang and J. S. Temenoff, "Engineering orthopedic tissue interfaces," Tissue Engineering Part B: Reviews, vol. 15, pp. 127-141, 2009.
  3. Martin, S. Miot, A. Barbero, M. Jakob, and D. Wendt, "Osteochondral tissue engineering," Journal of biomechanics, vol. 40, pp. 750-765, 2007.
  4. Gupta, M. Adhikary, M. Kumar, N. Bhardwaj, and B. B. Mandal, "Biomimetic, osteoconductive non-mulberry silk fiber reinforced tricomposite scaffolds for bone tissue engineering," ACS applied materials & interfaces, vol. 8, pp. 30797-30810, 2016.
  5. Wang, Y. Chu, J. He, W. Shao, Y. Zhou, K. Qi, et al., "A graded graphene oxide-hydroxyapatite/silk fibroin biomimetic scaffold for bone tissue engineering," Materials Science and Engineering: C, vol. 80, pp. 232-242, 2017.
  6. Nourmohammadi, F. Roshanfar, M. Farokhi, and M. H. Nazarpak, "Silk fibroin/kappa-carrageenan composite scaffolds with enhanced biomimetic mineralization for bone regeneration applications," Materials Science and Engineering: C, vol. 76, pp. 951-958, 2017.
  7. Melke, S. Midha, S. Ghosh, K. Ito, and S. Hofmann, "Silk fibroin as biomaterial for bone tissue engineering," Acta biomaterialia, vol. 31, pp. 1-16, 2016.
  8. L. Grayson, S. Bhumiratana, P. G. Chao, C. T. Hung, and G. Vunjak-Novakovic, "Spatial regulation of human mesenchymal stem cell differentiation in engineered osteochondral constructs: effects of pre-differentiation, soluble factors and medium perfusion," Osteoarthritis and cartilage, vol. 18, pp. 714-723, 2010.
  9. B. Yeatts, D. T. Choquette, and J. P. Fisher, "Bioreactors to influence stem cell fate: augmentation of mesenchymal stem cell signaling pathways via dynamic culture systems," Biochimica et Biophysica Acta (BBA)-General Subjects, vol. 1830, pp. 2470-2480, 2013.
  10. Pazzano, K. A. Mercier, J. M. Moran, S. S. Fong, D. D. DiBiasio, J. X. Rulfs, et al., "Comparison of chondrogensis in static and perfused bioreactor culture," Biotechnology progress, vol. 16, pp. 893-896, 2000.
  11. S. Tığlı, C. Cannizaro, M. Gümüşderelioğlu, and D. L. Kaplan, "Chondrogenesis in perfusion bioreactors using porous silk scaffolds and hESC‐derived MSCs," Journal of Biomedical Materials Research Part A, vol. 96, pp. 21-28, 2011.
  12. Zadegan, B. Vahidi, J. Nourmohammadi, and N. Haghighipour, "Biocompatibility and bioactivity behaviour of coelectrospun silk fibroin‐hydroxyapatite nanofibres using formic acid," Micro & Nano Letters, vol. 13, pp. 709-713, 2018.
  13. Klocke, M. Zeis, and L. Heidemanns, "Fluid structure interaction of thin graphite electrodes during flushing movements in sinking electrical discharge machining," CIRP Journal of Manufacturing Science and Technology, vol. 20, pp. 23-28, 2018.
  14. M. Murphy, M. G. Haugh, and F. J. O'brien, "The effect of mean pore size on cell attachment, proliferation and migration in collagen–glycosaminoglycan scaffolds for bone tissue engineering," Biomaterials, vol. 31, pp. 461-466, 2010.
  15. Zadegan, J. Nourmohammadi, B. Vahidi, and N. Haghighipour, "An investigation into osteogenic differentiation effects of silk fibroin-nettle (Urtica dioica L.) nanofibers," International journal of biological macromolecules, 133, 795-803, 2019.
  16. G. Haugh, C. M. Murphy, and F. J. O'Brien, "Novel freeze-drying methods to produce a range of collagen–glycosaminoglycan scaffolds with tailored mean pore sizes," Tissue Engineering Part C: Methods, vol. 16, pp. 887-894, 2010.
  17. Zhang, I. Hussain, M. Brust, M. F. Butler, S. P. Rannard, and A. I. Cooper, "Aligned two-and three-dimensional structures by directional freezing of polymers and nanoparticles," Nature materials, vol. 4, pp. 787-793, 2005.
  18. Zheng, F. Yang, S. Wang, S. Lu, W. Zhang, S. Liu, et al., "Fabrication and cell affinity of biomimetic structured PLGA/articular cartilage ECM composite scaffold," Journal of Materials Science: Materials in Medicine, vol. 22, pp. 693-704, 2011.
  19. Huang, R. D. Kamm, and R. T. Lee, "Cell mechanics and mechanotransduction: pathways, probes, and physiology," American Journal of Physiology-Cell Physiology, vol. 287, pp. C1-C11, 2004.
  20. J. Arnsdorf, P. Tummala, and C. R. Jacobs, "Non-canonical Wnt signaling and N-cadherin related β-catenin signaling play a role in mechanically induced osteogenic cell fate," PloS one, vol. 4, p. e5388, 2009.
  21. Nagel-Heyer, C. Goepfert, F. Feyerabend, J. P. Petersen, P. Adamietz, N. M. Meenen, et al., "Bioreactor cultivation of three-dimensional cartilage-carrier-constructs," Bioprocess and biosystems engineering, vol. 27, pp. 273-280, 2005.
  22. Woods, G. Wang, and F. Beier, "RhoA/ROCK signaling regulates Sox9 expression and actin organization during chondrogenesis," Journal of Biological Chemistry, vol. 280, pp. 11626-11634, 2005.
  23. J. Arnsdorf, P. Tummala, R. Y. Kwon, and C. R. Jacobs, "Mechanically induced osteogenic differentiation–the role of RhoA, ROCKII and cytoskeletal dynamics," Journal of cell science, vol. 122, pp. 546-553, 2009.
  24. Gonçalves, P. Costa, M. T. Rodrigues, I. R. Dias, R. L. Reis, and M. E. Gomes, "Effect of flow perfusion conditions in the chondrogenic differentiation of bone marrow stromal cells cultured onto starch based biodegradable scaffolds," Acta biomaterialia, vol. 7, pp. 1644-1652, 2011.
  25. Lefebvre and P. Smits, "Transcriptional control of chondrocyte fate and differentiation," Birth Defects Research Part C: Embryo Today: Reviews, vol. 75, pp. 200-212, 2005.
  26. J. Kelly and C. R. Jacobs, "The role of mechanical signals in regulating chondrogenesis and osteogenesis of mesenchymal stem cells," Birth Defects Research Part C: Embryo Today: Reviews, vol. 90, pp. 75-85, 2010.
Iranian Journal of Biomedical Engineering (IJBME)
Volume 16, Issue 3
Autumn 2022
Pages 271-280

  • Receive Date 13 February 2023
  • Revise Date 10 March 2023
  • Accept Date 12 March 2023

Home

Contact Us