Skip to main content
SpringerLink
Log in

Dynamics of tissue ingrowth in SIKVAV-modified highly superporous PHEMA scaffolds with oriented pores after bridging a spinal cord transection

  • Tissue Engineering Constructs and Cell Substrates
  • Original Research
  • Published:
Journal of Materials Science: Materials in Medicine Aims and scope Submit manuscript

Abstract

While many types of biomaterials have been evaluated in experimental spinal cord injury (SCI) research, little is known about the time-related dynamics of the tissue infiltration of these scaffolds. We analyzed the ingrowth of connective tissue, axons and blood vessels inside the superporous poly (2-hydroxyethyl methacrylate) hydrogel with oriented pores. The hydrogels, either plain or seeded with mesenchymal stem cells (MSCs), were implanted in spinal cord transection at the level of Th8. The animals were sacrificed at days 2, 7, 14, 28, 49 and 6 months after SCI and histologically evaluated. We found that within the first week, the hydrogels were already infiltrated with connective tissue and blood vessels, which remained stable for the next 6 weeks. Axons slowly and gradually infiltrated the hydrogel within the first month, after which the numbers became stable. Six months after SCI we observed rare axons crossing the hydrogel bridge and infiltrating the caudal stump. There was no difference in the tissue infiltration between the plain hydrogels and those seeded with MSCs. We conclude that while connective tissue and blood vessels quickly infiltrate the scaffold within the first week, axons show a rather gradual infiltration over the first month, and this is not facilitated by the presence of MSCs inside the hydrogel pores. Further research which is focused on the permissive micro-environment of the hydrogel scaffold is needed, to promote continuous and long-lasting tissue regeneration across the spinal cord lesion.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Geller HM, Fawcett JW. Building a bridge: engineering spinal cord repair. Exp Neurol. 2002;174:125–36. https://doi.org/10.1006/exnr.2002.7865.

    Article  Google Scholar 

  2. Hejčl AJ, Sameš P, Syková M. Experimental treatment of spinal cord injuries. Cesk Slov Neurol N. 2015;78/111:377–93.

    Article  Google Scholar 

  3. Hejcl A, Sedy J, Kapcalova M, Toro DA, Amemori T, Lesny P, et al. HPMA-RGD hydrogels seeded with mesenchymal stem cells improve functional outcome in chronic spinal cord injury. Stem Cells Dev. 2010;19:1535–46. https://doi.org/10.1089/scd.2009.0378.

    Article  CAS  Google Scholar 

  4. Hejcl A, Ruzicka J, Kapcalova M, Turnovcova K, Krumbholcova E, Pradny M, et al. Adjusting the chemical and physical properties of hydrogels leads to improved stem cell survival and tissue ingrowth in spinal cord injury reconstruction: a comparative study of four methacrylate hydrogels. Stem Cells Dev. 2013;22:2794–805. https://doi.org/10.1089/scd.2012.0616.

    Article  CAS  Google Scholar 

  5. Nomura H, Baladie B, Katayama Y, Morshead CM, Shoichet MS, Tator CH. Delayed implantation of intramedullary chitosan channels containing nerve grafts promotes extensive axonal regeneration after spinal cord injury. Neurosurgery. 2008;63:127–41. https://doi.org/10.1227/01.NEU.0000335080.47352.31.

    Article  Google Scholar 

  6. Kubinova S, Horak D, Hejcl A, Plichta Z, Kotek J, Proks V, et al. SIKVAV-modified highly superporous PHEMA scaffolds with oriented pores for spinal cord injury repair. J Tissue Eng Regen Med. 2015;9:1298–309. https://doi.org/10.1002/term.1694.

    Article  CAS  Google Scholar 

  7. Hejcl A, Lesny P, Pradny M, Sedy J, Zamecnik J, Jendelova P, et al. Macroporous hydrogels based on 2-hydroxyethyl methacrylate. Part 6: 3D hydrogels with positive and negative surface charges and polyelectrolyte complexes in spinal cord injury repair. J Mater Sci Mater Med. 2009;20:1571–7. https://doi.org/10.1007/s10856-009-3714-4.

    Article  CAS  Google Scholar 

  8. Kubinova S, Horak D, Kozubenko N, Vanecek V, Proks V, Price J, et al. The use of superporous Ac-CGGASIKVAVS-OH-modified PHEMA scaffolds to promote cell adhesion and the differentiation of human fetal neural precursors. Biomaterials. 2010;31:5966–75. https://doi.org/10.1016/j.biomaterials.2010.04.040.

    Article  CAS  Google Scholar 

  9. Salek P, Korecka L, Horak D, Petrovsky E, Kovarova J, Metelka R, et al. Immunomagnetic sulfonated hypercrosslinked polystyrene microspheres for electrochemical detection of proteins. J Mater Chem. 2011;21:14783–92. https://doi.org/10.1039/c1jm12475g.

    Article  CAS  Google Scholar 

  10. Okabe M, Ikawa M, Kominami K, Nakanishi T, Nishimune Y. ‘Green mice’ as a source of ubiquitous green cells. FEBS Lett. 1997;407:313–9.

    Article  CAS  Google Scholar 

  11. Hejcl A, Urdzikova L, Sedy J, Lesny P, Pradny M, Michalek J, et al. Acute and delayed implantation of positively charged 2-hydroxyethyl methacrylate scaffolds in spinal cord injury in the rat. J Neurosurg Spine. 2008;8:67–73. https://doi.org/10.3171/SPI-08/01/067.

    Article  Google Scholar 

  12. Horák D, Hradil H, Lapčíková M, Šlouf M. Superporous poly(2-hydroxyethyl methacrylate) based scaffolds: preparation and characterization. Polymer. 2008;49:2046–54.

    Article  Google Scholar 

  13. Horak D, Kroupova J, Slouf M, Dvorak P. Poly(2-hydroxyethyl methacrylate)-based slabs as a mouse embryonic stem cell support. Biomaterials. 2004;25:5249–60. https://doi.org/10.1016/j.biomaterials.2003.12.031.

    Article  CAS  Google Scholar 

  14. Fitch MT, Doller C, Combs CK, Landreth GE, Silver J. Cellular and molecular mechanisms of glial scarring and progressive cavitation: in vivo and in vitro analysis of inflammation-induced secondary injury after CNS trauma. J Neurosci. 1999;19:8182–98.

    Article  CAS  Google Scholar 

  15. Ruzicka J, Romanyuk N, Hejcl A, Vetrik M, Hruby M, Cocks G, et al. Treating spinal cord injury in rats with a combination of human fetal neural stem cells and hydrogels modified with serotonin. Acta Neurobiol Exp. 2013;73:102–15.

    Google Scholar 

  16. Hejcl A, Lesny P, Pradny M, Michalek J, Jendelova P, Stulik J, et al. Biocompatible hydrogels in spinal cord injury repair. Physiol Res. 2008;57(Suppl 3):S121–32.

    CAS  Google Scholar 

  17. Amr SM, Gouda A, Koptan WT, Galal AA, Abdel-Fattah DS, Rashed LA, et al. Bridging defects in chronic spinal cord injury using peripheral nerve grafts combined with a chitosan-laminin scaffold and enhancing regeneration through them by co-transplantation with bone-marrow-derived mesenchymal stem cells: case series of 14 patients. J Spinal Cord Med. 2014;37:54–71. https://doi.org/10.1179/2045772312Y.0000000069.

    Article  Google Scholar 

  18. Li N, Sarojini H, An J, Wang E. Prosaposin in the secretome of marrow stroma-derived neural progenitor cells protects neural cells from apoptotic death. J Neurochem. 2010;112:1527–38. https://doi.org/10.1111/j.1471-4159.2009.06565.x.

    Article  CAS  Google Scholar 

  19. Hao P, Liang Z, Piao H, Ji X, Wang Y, Liu Y, et al. Conditioned medium of human adipose-derived mesenchymal stem cells mediates protection in neurons following glutamate excitotoxicity by regulating energy metabolism and GAP-43 expression. Metab Brain Dis. 2014;29:193–205. https://doi.org/10.1007/s11011-014-9490-y.

    Article  CAS  Google Scholar 

  20. Gunther MI, Weidner N, Muller R, Blesch A. Cell-seeded alginate hydrogel scaffolds promote directed linear axonal regeneration in the injured rat spinal cord. Acta Biomater. 2015;27:140–50. https://doi.org/10.1016/j.actbio.2015.09.001.

    Article  Google Scholar 

  21. Oliveira E, Assuncao-Silva RC, Ziv-Polat O, Gomes ED, Teixeira FG, Silva NA, et al. Influence of different ECM-like hydrogels on neurite outgrowth induced by adipose tissue-derived stem cells. Stem Cells Int. 2017;2017:6319129 https://doi.org/10.1155/2017/6319129.

    Article  CAS  Google Scholar 

  22. Papa S, Vismara I, Mariani A, Barilani M, Rimondo S, De Paola M, et al. Mesenchymal stem cells encapsulated into biomimetic hydrogel scaffold gradually release CCL2 chemokine in situ preserving cytoarchitecture and promoting functional recovery in spinal cord injury. J Control Release. 2018;278:49–56. https://doi.org/10.1016/j.jconrel.2018.03.034.

    Article  CAS  Google Scholar 

  23. Qu J, Zhang H. Roles of mesenchymal stem cells in spinal cord injury. Stem Cells Int. 2017;2017:5251313 https://doi.org/10.1155/2017/5251313.

    Article  Google Scholar 

  24. Stewart AN, Matyas JJ, Welchko RM, Goldsmith AD, Zeiler SE, Hochgeschwender U, et al. SDF-1 overexpression by mesenchymal stem cells enhances GAP-43-positive axonal growth following spinal cord injury. Restor Neurol Neurosci. 2017;35:395–411. https://doi.org/10.3233/RNN-160678.

    Article  CAS  Google Scholar 

  25. Yang EZ, Zhang GW, Xu JG, Chen S, Wang H, Cao LL, et al. Multichannel polymer scaffold seeded with activated Schwann cells and bone mesenchymal stem cells improves axonal regeneration and functional recovery after rat spinal cord injury. Acta Pharmacol Sin. 2017;38:623–37. https://doi.org/10.1038/aps.2017.11.

    Article  CAS  Google Scholar 

  26. Tashiro K, Sephel GC, Weeks B, Sasaki M, Martin GR, Kleinman HK, et al. A synthetic peptide containing the IKVAV sequence from the A chain of laminin mediates cell attachment, migration, and neurite outgrowth. J Biol Chem. 1989;264:16174–82.

    CAS  Google Scholar 

  27. Tysseling-Mattiace VM, Sahni V, Niece KL, Birch D, Czeisler C, Fehlings MG, et al. Self-assembling nanofibers inhibit glial scar formation and promote axon elongation after spinal cord injury. J Neurosci. 2008;28:3814–23. https://doi.org/10.1523/JNEUROSCI.0143-08.2008.

    Article  CAS  Google Scholar 

  28. Tysseling VM, Sahni V, Pashuck ET, Birch D, Hebert A, Czeisler C, et al. Self-assembling peptide amphiphile promotes plasticity of serotonergic fibers following spinal cord injury. J Neurosci Res. 2010;88:3161–70. https://doi.org/10.1002/jnr.22472.

    Article  CAS  Google Scholar 

  29. Andrews MR, Czvitkovich S, Dassie E, Vogelaar CF, Faissner A, Blits B, et al. Alpha9 integrin promotes neurite outgrowth on tenascin-C and enhances sensory axon regeneration. J Neurosci. 2009;29:5546–57. https://doi.org/10.1523/JNEUROSCI.0759-09.2009.

    Article  CAS  Google Scholar 

  30. Cheah M, Andrews MR, Chew DJ, Moloney EB, Verhaagen J, Fassler R, et al. Expression of an activated integrin promotes long-distance sensory axon regeneration in the spinal cord. J Neurosci. 2016;36:7283–97. https://doi.org/10.1523/JNEUROSCI.0901-16.2016.

    Article  CAS  Google Scholar 

  31. Deumens R, Koopmans GC, Honig WM, Hamers FP, Maquet V, Jerome R, et al. Olfactory ensheathing cells, olfactory nerve fibroblasts and biomatrices to promote long-distance axon regrowth and functional recovery in the dorsally hemisected adult rat spinal cord. Exp Neurol. 2006;200:89–103. https://doi.org/10.1016/j.expneurol.2006.01.030.

    Article  CAS  Google Scholar 

  32. Deumens R, Koopmans GC, Honig WM, Maquet V, Jerome R, Steinbusch HW, et al. Limitations in transplantation of astroglia-biomatrix bridges to stimulate corticospinal axon regrowth across large spinal lesion gaps. Neurosci Lett. 2006;400:208–12. https://doi.org/10.1016/j.neulet.2006.02.050.

    Article  CAS  Google Scholar 

  33. Deng LX, Deng P, Ruan Y, Xu ZC, Liu NK, Wen X, et al. A novel growth-promoting pathway formed by GDNF-overexpressing Schwann cells promotes propriospinal axonal regeneration, synapse formation, and partial recovery of function after spinal cord injury. J Neurosci. 2013;33:5655–67. https://doi.org/10.1523/JNEUROSCI.2973-12.2013.

    Article  CAS  Google Scholar 

  34. Iida T, Nakagawa M, Asano T, Fukushima C, Tachi K. Free vascularized lateral femoral cutaneous nerve graft with anterolateral thigh flap for reconstruction of facial nerve defects. J Reconstr Microsurg. 2006;22:343–8. https://doi.org/10.1055/s-2006-946711.

    Article  Google Scholar 

  35. Glaser J, Gonzalez R, Sadr E, Keirstead HS. Neutralization of the chemokine CXCL10 reduces apoptosis and increases axon sprouting after spinal cord injury. J Neurosci Res. 2006;84:724–34. https://doi.org/10.1002/jnr.20982.

    Article  CAS  Google Scholar 

  36. Gao XR, Xu HJ, Wang LF, Liu CB, Yu F. Mesenchymal stem cell transplantation carried in SVVYGLR modified self-assembling peptide promoted cardiac repair and angiogenesis after myocardial infarction. Biochem Biophys Res Commun. 2017;491:112–8. https://doi.org/10.1016/j.bbrc.2017.07.056.

    Article  CAS  Google Scholar 

  37. Hou Y, Ryu CH, Jun JA, Kim SM, Jeong CH, Jeun SS. IL-8 enhances the angiogenic potential of human bone marrow mesenchymal stem cells by increasing vascular endothelial growth factor. Cell Biol Int. 2014;38:1050–9. https://doi.org/10.1002/cbin.10294.

    Article  CAS  Google Scholar 

  38. Wang C, Poon S, Murali S, Koo CY, Bell TJ, Hinkley SF, et al. Engineering a vascular endothelial growth factor 165-binding heparan sulfate for vascular therapy. Biomaterials. 2014;35:6776–86. https://doi.org/10.1016/j.biomaterials.2014.04.084.

    Article  CAS  Google Scholar 

  39. Golebiewska EM, Poole AW. Platelet secretion: from haemostasis to wound healing and beyond. Blood Rev. 2015;29:153–62. https://doi.org/10.1016/j.blre.2014.10.003.

    Article  CAS  Google Scholar 

  40. Ullm S, Kruger A, Tondera C, Gebauer TP, Neffe AT, Lendlein A, et al. Biocompatibility and inflammatory response in vitro and in vivo to gelatin-based biomaterials with tailorable elastic properties. Biomaterials. 2014;35:9755–66. https://doi.org/10.1016/j.biomaterials.2014.08.023.

    Article  CAS  Google Scholar 

  41. Feng Y, Li Q, Wu D, Niu Y, Yang C, Dong L, et al. A macrophage-activating, injectable hydrogel to sequester endogenous growth factors for in situ angiogenesis. Biomaterials. 2017;134:128–42. https://doi.org/10.1016/j.biomaterials.2017.04.042.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We would like to thank Frances Zatřepálková and Jan Lodin for proofreading the manuscript. The study has been supported by 2 grants from the Grant Agency of the Czech Republic 14-14961S, 17-11140S, the Operational Programme Research, Development and Education in the framework of the project “Centre of Reconstructive Neuroscience “, registration number CZ.02.1.01/0.0./0.0/15_003/0000419 and by the grant from the Ministry of Education, Youth and Sports No. LO1309.

Author information

Authors and Affiliations

  1. Institute of Experimental Medicine, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague, Czech Republic

    Aleš Hejčl, Jiří Růžička, Šárka Kubinová & Pavla Jendelová

  2. Department of Neurosurgery, J. E. Purkinje University, Masaryk Hospital, Sociální péče 12A, 401 13, Ústí nad Labem, Czech Republic

    Aleš Hejčl

  3. Department of Neuroscience, 2nd Faculty of Medicine, Charles University, V Úvalu 84, 150 06, Prague 5, Czech Republic

    Jiří Růžička & Pavla Jendelová

  4. Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Heyrovského nám.2, 162 06, Praha 6, Břevnov, Czech Republic

    Vladimír Proks, Hana Macková & Daniel Horák

  5. Department of Neurosurgery, Motol University Hospital, V Úvalu 84, Prague 5, 150 06, Czech Republic

    Dmitry Tukmachev

  6. Department of Mathematics, Faculty of Science, J. E. Purkyně University, České mládeže 8, 400 96, Ústí nad Labem, Czech Republic

    Jiří Cihlář

Authors
  1. Aleš Hejčl
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jiří Růžička
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Vladimír Proks
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hana Macková
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Šárka Kubinová
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Dmitry Tukmachev
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jiří Cihlář
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Daniel Horák
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Pavla Jendelová
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Aleš Hejčl.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hejčl, A., Růžička, J., Proks, V. et al. Dynamics of tissue ingrowth in SIKVAV-modified highly superporous PHEMA scaffolds with oriented pores after bridging a spinal cord transection. J Mater Sci: Mater Med 29, 89 (2018). https://doi.org/10.1007/s10856-018-6100-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s10856-018-6100-2

Search

Navigation