Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Guiding intracortical brain tumour cells to an extracortical cytotoxic hydrogel using aligned polymeric nanofibres

Abstract

Glioblastoma multiforme is an aggressive, invasive brain tumour with a poor survival rate. Available treatments are ineffective and some tumours remain inoperable because of their size or location. The tumours are known to invade and migrate along white matter tracts and blood vessels. Here, we exploit this characteristic of glioblastoma multiforme by engineering aligned polycaprolactone (PCL)-based nanofibres for tumour cells to invade and, hence, guide cells away from the primary tumour site to an extracortical location. This extracortial sink is a cyclopamine drug-conjugated, collagen-based hydrogel. When aligned PCL-nanofibre films in a PCL/polyurethane carrier conduit were inserted in the vicinity of an intracortical human U87MG glioblastoma xenograft, a significant number of human glioblastoma cells migrated along the aligned nanofibre films and underwent apoptosis in the extracortical hydrogel. Tumour volume in the brain was significantly lower following insertion of aligned nanofibre implants compared with the application of smooth fibres or no implants.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

Figure 1: Schematic and image of conduit inserted into a rat brain.
Figure 2: In vitro tumour cell migration on aligned nanofibre films versus smooth films.
Figure 3: Live/Dead assay for cell viability in hydrogels.
Figure 4: Glioblastoma cells in the conduit in vivo.
Figure 5: Proliferation of U87MG-eGFP cells in the brain and conduits.
Figure 6: Apoptosis staining of glioblastoma cells in the cyclopamine hydrogel-filled conduits.

References

  1. 1

    Davis, F. G., Freels, S., Grutsch, J., Barlas, S. & Brem, S. Survival rates in patients with primary malignant brain tumors stratified by patient age and tumor histological type: an analysis based on Surveillance, Epidemiology, and End Results (SEER) data, 1973–1991. J. Neurosurg. 88, 1–10 (1998).

    CAS  Article  Google Scholar 

  2. 2

    Wiesner, S. M., Freese, A. & Ohlfest, J. R. Emerging concepts in glioma biology: implications for clinical protocols and rational treatment strategies. Neurosurg. Focus 19, E3 (2005).

    Article  Google Scholar 

  3. 3

    Tuettenberg, J. et al. Recurrence pattern in glioblastoma multiforme patients treated with anti-angiogenic chemotherapy. J. Cancer Res. Clin. Oncol. 135, 1239–1244 (2009).

    CAS  Article  Google Scholar 

  4. 4

    Rostomily, R. C. et al. Multimodality management of recurrent adult malignant gliomas: results of a phase II multiagent chemotherapy study and analysis of cytoreductive surgery. Neurosurgery 35, 378–388 (1994).

    CAS  Article  Google Scholar 

  5. 5

    Bar, E. E. et al. Cyclopamine-mediated hedgehog pathway inhibition depletes stem-like cancer cells in glioblastoma. Stem Cells 25, 2524–2533 (2007).

    CAS  Article  Google Scholar 

  6. 6

    Bellail, A. C., Hunter, S. B., Brat, D. J., Tan, C. & Van Meir, E. G. Microregional extracellular matrix heterogeneity in brain modulates glioma cell invasion. Int. J. Biochem. Cell Biol. 36, 1046–1069 (2004).

    CAS  Article  Google Scholar 

  7. 7

    Bernstein, J. J. & Woodard, C. A. Glioblastoma cells do not intravasate into blood vessels. Neurosurgery 36, 124–132 (1995).

    CAS  Article  Google Scholar 

  8. 8

    Weiner, H. L. et al. Induction of medulloblastomas in mice by sonic hedgehog, independent of Gli1. Cancer Res. 62, 6385–6389 (2002).

    CAS  Google Scholar 

  9. 9

    Becher, O. J. et al. Gli activity correlates with tumor grade in platelet-derived growth factor-induced gliomas. Cancer Res. 68, 2241–2249 (2008).

    CAS  Article  Google Scholar 

  10. 10

    Grossman, S. A. et al. The intracerebral distribution of BCNU delivered by surgically implanted biodegradable polymers. J. Neurosurg. 76, 640–647 (1992).

    CAS  Article  Google Scholar 

  11. 11

    Kohn, K. W. Interstrand cross-linking of DNA by 1,3-bis(2-chloroethyl)-1-nitrosourea and other 1-(2-haloethyl)-1-nitrosoureas. Cancer Res. 37, 1450–1454 (1977).

    CAS  Google Scholar 

  12. 12

    Kee, N., Sivalingam, S., Boonstra, R. & Wojtowicz, J. M. The utility of Ki-67 and BrdU as proliferative markers of adult neurogenesis. J. Neurosci. Methods 115, 97–105 (2002).

    CAS  Article  Google Scholar 

  13. 13

    Haghparast, S. M. A., Kihara, T., Shimizu, Y., Yuba, S. & Miyake, J. Actin-based biomechanical features of suspended normal and cancer cells. J. Biosci. Bioeng. 116, 380–385 (2013).

    CAS  Article  Google Scholar 

  14. 14

    Biran, R., Noble, M. D. & Tresco, P. A. Directed nerve outgrowth is enhanced by engineered glial substrates. Exp. Neurol. 184, 141–152 (2003).

    CAS  Article  Google Scholar 

  15. 15

    Kanda, S. et al. Sonic hedgehog induces capillary morphogenesis by endothelial cells through phosphoinositide 3-kinase. J. Biol. Chem. 278, 8244–8249 (2003).

    CAS  Article  Google Scholar 

  16. 16

    Baluk, P., Morikawa, S., Haskell, A., Mancuso, M. & McDonald, D. M. Abnormalities of basement membrane on blood vessels and endothelial sprouts in tumors. Am. J. Pathol. 163, 1801–1815 (2003).

    Article  Google Scholar 

  17. 17

    Hernandez, L., Smirnova, T., Wyckoff, J., Condeelis, J. & Segall, J. E. In vivo assay for tumor cell invasion. Methods Mol. Biol. 571, 227–238 (2009).

    CAS  Article  Google Scholar 

  18. 18

    Wyckoff, J., Gligoijevic, B., Entenberg, D., Segall, J. E. & Condeelis, J. The in vivo invasion assay: preparation and handling of collection needles. Cold Spring Harbor Protocols 10, 1232–1234 (2011).

    Google Scholar 

  19. 19

    Kim, Y. T., Haftel, V. K., Kumar, S. & Bellamkonda, R. V. The role of aligned polymer fiber-based constructs in the bridging of long peripheral nerve gaps. Biomaterials 29, 3117–3127 (2008).

    CAS  Article  Google Scholar 

  20. 20

    Karumbaiah, L. et al. The upregulation of specific interleukin (IL) receptor antagonists and paradoxical enhancement of neuronal apoptosis due to electrode induced strain and brain micromotion. Biomaterials 33, 5983–5996 (2012).

    CAS  Article  Google Scholar 

Download references

Acknowledgements

We would like to thank J. Leisen for his technical expertise with the MR scanner and J. Lyon and S. Nayebosadri for their assistance with the animal surgeries and immunohistochemistry. We would also like to acknowledge funding support for this project from National Institutes of Health EUREKA R01 CA153229 (NCI) (R.V.B.), the Georgia Research Alliance (R.V.B.), and Ian’s Friends Foundation (R.V.B.)

Author information

Affiliations

Authors

Contributions

R.V.B. conceived the tumour ‘exvasion’ strategy using nanofibre films towards an apoptosis-inducing hydrogel and was responsible for the overall study design. A.J. implemented the above strategy by means of the design of a nanofibre tumour guide and a hydrogel sink, and also performed the surgeries, designed the experiments and analysed the results. M.B. oversaw the smooth film control studies, as well as quantified the Ki-67+ staining and tumour volume. G.D.P. fabricated the polymer conduits and nanofibre films. C.M.V. designed and performed the collagen and cyclopamine collagen two-well depot Live/Dead experiments, based on a cyclopamine strategy suggested by T.J.M. V.J.M. assisted with animal surgeries and tissue sectioning. A.V. performed haematoxylin and eosin staining and helped with histological assessment. S.B.P. stably transfected the U87MG cells to express eGFP. B.B helped with surgical strategy.

Corresponding author

Correspondence to Ravi V. Bellamkonda.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Information (PDF 1363 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Jain, A., Betancur, M., Patel, G. et al. Guiding intracortical brain tumour cells to an extracortical cytotoxic hydrogel using aligned polymeric nanofibres. Nature Mater 13, 308–316 (2014). https://doi.org/10.1038/nmat3878

Download citation

Further reading

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing