Abstract
Tissue engineered nerve grafts (TENGs) are considered a promising alternative to autologous nerve grafting, which is considered the “gold standard” clinical strategy for peripheral nerve repair. Here, we immobilized tumor necrosis factor-α (TNF-α) inhibitors onto a nerve conduit, which was introduced into a chitosan (CS) matrix scaffold utilizing genipin (GP) as the crosslinking agent, to fabricate CS-GP-TNF-α inhibitor nerve conduits. The in vitro release kinetics of TNF-α inhibitors from the CS-GP-TNF-α inhibitor nerve conduits were investigated using high-performance liquid chromatography. The in vivo continuous release profile of the TNF-α inhibitors released from the CS-GP-TNF-α inhibitor nerve conduits was measured using an enzyme-linked immunosorbent assay over 14 days. We found that the amount of TNF-α inhibitors released decreased with time after the bridging of the sciatic nerve defects in rats. Moreover, 4 and 12 weeks after surgery, histological analyses and functional evaluations were carried out to assess the influence of the TENG on regeneration. Immunochemistry performed 4 weeks after grafting to assess early regeneration outcomes revealed that the TENG strikingly promoted axonal outgrowth. Twelve weeks after grafting, the TENG accelerated myelin sheath formation, as well as functional restoration. In general, the regenerative outcomes following TENG more closely paralleled findings observed with autologous grafting than the use of the CS matrix scaffold. Collectively, our data indicate that the CS-GP-TNF-α inhibitor nerve conduits comprised an elaborate system for sustained release of TNF-α inhibitors in vitro, while studies in vivo demonstrated that the TENG could accelerate regenerating axonal outgrowth and functional restoration. The introduction of CS-GP-TNF-α-inhibitor nerve conduits into a scaffold may contribute to an efficient and adaptive immune microenvironment that can be used to facilitate peripheral nerve repair.
Similar content being viewed by others
References
Akerman, P., P. Cote, S. Q. Yang, C. McClain, S. Nelson, G. J. Bagby, et al. Antibodies to tumor necrosis factor-alpha inhibit liver regeneration after partial hepatectomy. Am. J. Physiol. 263:G579–G585, 1992.
Avellino, A. M., D. Hart, A. T. Dailey, M. MacKinnon, D. Ellegala, and M. Kliot. Differential macrophage responses in the peripheral and central nervous system during wallerian degeneration of axons. Exp. Neurol. 136:183–198, 1995.
Battiston, B., P. Titolo, D. Ciclamini, and B. Panero. Peripheral nerve defects: overviews of practice in Europe. Hand Clin. 33:545–550, 2017.
Chiono, V., and C. Tonda-Turo. Trends in the design of nerve guidance channels in peripheral nerve tissue engineering. Prog. Neurobiol. 131:87–104, 2015.
Cressman, D. E., L. E. Greenbaum, R. A. DeAngelis, G. Ciliberto, E. E. Furth, V. Poli, et al. Liver failure and defective hepatocyte regeneration in interleukin-6-deficient mice. Science. 274:1379–1383, 1996.
Dalamagkas, K., M. Tsintou, and A. Seifalian. Advances in peripheral nervous system regenerative therapeutic strategies: a biomaterials approach. Mater. Sci. Eng. C. 65:425–432, 2016.
Freeman, M. R. Signaling mechanisms regulating Wallerian degeneration. Curr. Opin. Neurobiol. 27:224–231, 2014.
Gnavi, S., C. Barwig, T. Freier, K. Haastert-Talini, C. Grothe, and S. Geuna. The use of chitosan-based scaffolds to enhance regeneration in the nervous system. Int. Rev. Neurobiol. 109:1–62, 2013.
Gomez-Sanchez, J. A., L. Carty, M. Iruarrizaga-Lejarreta, M. Palomo-Irigoyen, M. Varela-Rey, M. Griffith, et al. Schwann cell autophagy, myelinophagy, initiates myelin clearance from injured nerves. J. Cell Biol. 210:153–168, 2015.
Gu, X., F. Ding, and D. F. Williams. Neural tissue engineering options for peripheral nerve regeneration. Biomaterials. 35:6143–6156, 2014.
Gu, X., F. Ding, Y. Yang, and J. Liu. Construction of tissue engineered nerve grafts and their application in peripheral nerve regeneration. Prog. Neurobiol. 93:204–230, 2011.
Gurtner, G. C., S. Werner, Y. Barrandon, and M. T. Longaker. Wound repair and regeneration. Nature. 453:314–321, 2008.
He, Z., and Y. Jin. Intrinsic control of axon regeneration. Neuron. 90:437–451, 2016.
Huebner, E. A., and S. M. Strittmatter. Axon regeneration in the peripheral and central nervous systems. Results Probl. Cell Differ. 48:339–351, 2009.
Jang, S. Y., B. A. Yoon, Y. K. Shin, S. H. Yun, Y. R. Jo, Y. Y. Choi, et al. Schwann cell dedifferentiation-associated demyelination leads to exocytotic myelin clearance in inflammatory segmental demyelination. Glia. 65:1848–1862, 2017.
Karin, M., and H. Clevers. Reparative inflammation takes charge of tissue regeneration. Nature. 529:307–315, 2016.
Koppes, R. A., S. Park, T. Hood, X. Jia, N. Abdolrahim Poorheravi, A. H. Achyuta, et al. Thermally drawn fibers as nerve guidance scaffolds. Biomaterials. 81:27–35, 2016.
Leung, L., and C. M. Cahill. TNF-alpha and neuropathic pain–a review. J. Neuroinflamm. 7:27, 2010.
Li, S., C. Xue, Y. Yuan, R. Zhang, Y. Wang, Y. Wang, et al. The transcriptional landscape of dorsal root ganglia after sciatic nerve transection. Sci. Rep. 5:16888, 2015.
Liu, Y., L. J. Zhou, J. Wang, D. Li, W. J. Ren, J. Peng, et al. TNF-alpha differentially regulates synaptic plasticity in the hippocampus and spinal cord by microglia-dependent mechanisms after peripheral nerve injury. J. Neurosci. 37:871–881, 2017.
Martini, R., S. Fischer, R. Lopez-Vales, and S. David. Interactions between Schwann cells and macrophages in injury and inherited demyelinating disease. Glia. 56:1566–1577, 2008.
Mc Guire, C., R. Beyaert, and G. van Loo. Death receptor signalling in central nervous system inflammation and demyelination. Trends Neurosci. 34:619–628, 2011.
Pabari, A., S. Y. Yang, A. M. Seifalian, and A. Mosahebi. Modern surgical management of peripheral nerve gap. JPRAS. 63:1941–1948, 2010.
Pateman, C. J., A. J. Harding, A. Glen, C. S. Taylor, C. R. Christmas, P. P. Robinson, et al. Nerve guides manufactured from photocurable polymers to aid peripheral nerve repair. Biomaterials. 49:77–89, 2015.
Raimondo, S., M. Fornaro, P. Tos, B. Battiston, M. G. Giacobini-Robecchi, and S. Geuna. Perspectives in regeneration and tissue engineering of peripheral nerves. Ann. Anat. 193:334–340, 2011.
Sadtler, K., K. Estrellas, B. W. Allen, M. T. Wolf, H. Fan, A. J. Tam, et al. Developing a pro-regenerative biomaterial scaffold microenvironment requires T helper 2 cells. Science. 352:366–370, 2016.
Sivashankari, P. R., and M. Prabaharan. Prospects of chitosan-based scaffolds for growth factor release in tissue engineering. Int. J. Biol. Macromol. 93:1382–1389, 2016.
Spivey, E. C., Z. Z. Khaing, J. B. Shear, and C. E. Schmidt. The fundamental role of subcellular topography in peripheral nerve repair therapies. Biomaterials. 33:4264–4276, 2012.
Tang, X., H. Qin, X. Gu, and X. Fu. China’s landscape in regenerative medicine. Biomaterials. 124:78–94, 2017.
Tang, X., Y. Wang, S. Zhou, T. Qian, and X. Gu. Signaling pathways regulating dose-dependent dual effects of TNF-alpha on primary cultured Schwann cells. Mol. Cell Biochem. 378:237–246, 2013.
Tang, X., C. Xue, Y. Wang, F. Ding, Y. Yang, and X. Gu. Bridging peripheral nerve defects with a tissue engineered nerve graft composed of an in vitro cultured nerve equivalent and a silk fibroin-based scaffold. Biomaterials. 33:3860–3867, 2012.
Tzekova, N., A. Heinen, S. Bunk, C. Hermann, H. P. Hartung, B. Reipert, et al. Immunoglobulins stimulate cultured Schwann cell maturation and promote their potential to induce axonal outgrowth. J. Neuroinflamm. 12:107, 2015.
Vargas, M. E., J. Watanabe, S. J. Singh, W. H. Robinson, and B. A. Barres. Endogenous antibodies promote rapid myelin clearance and effective axon regeneration after nerve injury. PNAS. 107:11993–11998, 2010.
Wang, Y., X. Tang, B. Yu, Y. Gu, Y. Yuan, D. Yao, et al. Gene network revealed involvements of Birc2, Birc3 and Tnfrsf1a in anti-apoptosis of injured peripheral nerves. PLoS ONE. 7:e43436, 2012.
Wong, K. M., E. Babetto, and B. Beirowski. Axon degeneration: make the Schwann cell great again. Neural Regen. Res. 12:518–524, 2017.
Yang, Y., W. Zhao, J. He, Y. Zhao, F. Ding, and X. Gu. Nerve conduits based on immobilization of nerve growth factor onto modified chitosan by using genipin as a crosslinking agent. Eur. J. Pharm. Biopharm. 79:519–525, 2011.
Yi, S., X. Tang, J. Yu, J. Liu, F. Ding, and X. Gu. Microarray and qPCR analyses of wallerian degeneration in rat sciatic nerves. Front. Cell. Neurosci. 11:22, 2017.
Zhao, Y., Y. Wang, J. Gong, L. Yang, C. Niu, X. Ni, et al. Chitosan degradation products facilitate peripheral nerve regeneration by improving macrophage-constructed microenvironments. Biomaterials. 134:64–77, 2017.
Acknowledgments
We appreciate the financial support from the National Key Research and Development Program of China (No. 2016YFC1101603), National Natural Science Foundation of China (Nos. 81370043 and 81671823), and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).
Author Contributions
XT and YY designed the research; LZ, WZ, CN, YZ, HS and XT performed the experiments; LZ, CN, YZ, HS, YW, YY, and XT analyzed data; XT wrote the paper.
Conflict of interest
The authors declare no conflicts of interest.
Author information
Authors and Affiliations
Corresponding authors
Additional information
Associate Editor Michael Gower oversaw the review of this article.
Rights and permissions
About this article
Cite this article
Zhang, L., Zhao, W., Niu, C. et al. Genipin-Cross-Linked Chitosan Nerve Conduits Containing TNF-α Inhibitors for Peripheral Nerve Repair. Ann Biomed Eng 46, 1013–1025 (2018). https://doi.org/10.1007/s10439-018-2011-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10439-018-2011-0