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Mechanisms of Reduced Astrocyte Surface Coverage in Cortical Neuron-Glia Co-cultures on Nanoporous Gold Surfaces

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Abstract

Nanoporous gold (np-Au) is a promising multifunctional material for neural electrodes. We have previously shown that np-Au nanotopography reduces astrocyte surface coverage (linked to undesirable gliosis) while maintaining high neuronal coverage in a cortical primary neuron-glia co-culture model as long as 2 weeks in vitro. Here, we investigate the potential influence of secreted soluble factors from cells grown on np-Au leading to the cell type-specific surface coverage on conventional tissue culture plastic. We then test the hypothesis that secretion of factors is responsible for inhibiting astrocyte coverage on np-Au. In order to assess whether factors secreted from cells grown on np-Au surfaces reduced surface coverage by astrocytes, we seeded fresh primary rat neuron-glia co-cultures on conventional polystyrene culture dishes, but maintained the cells in conditioned media from co-cultures grown on np-Au surfaces. After 1 week in vitro, a preferential reduction in astrocyte surface coverage was not observed, suggesting that soluble factors are not playing a role. In contrast, 4 h after cell seeding there were a significant number of non-adhered, yet still viable, cells for the cultures on np-Au surfaces. We hypothesize that the non-adherent cells are mainly astrocytes, because: (i) there was no difference in neuronal cell coverage between np-Au and pl-Au for long culture durations and (ii) neurons are post-mitotic and not expected to increase in number upon attaching to the surface. Overall, the results suggest that the np-Au topography leads to preferential neuronal attachment shortly after cell seeding and limits astrocyte-specific np-Au surface coverage at longer culture durations.

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References

  1. Agarwal, A., and D. E. Bergles. Astrocyte morphology is controlled by neuron-derived FGF. Neuron 83:255–257, 2014.

    Article  Google Scholar 

  2. Arregui, C., S. Carbonetto, and L. McKerracher. Characterization of neural cell adhesion sites: point contacts are the sites of interaction between integrins and the cytoskeleton in PC12 cells. J. Neurosci. 14:6967–6977, 1994.

    Google Scholar 

  3. Biggs, M. J. P., R. G. Richards, and M. J. Dalby. Nanotopographical modification: a regulator of cellular function through focal adhesions. Nanomedicine 6:619–633, 2010.

    Google Scholar 

  4. Blumenthal, N. R., O. Hermanson, B. Heimrich, and V. P. Shastri. Stochastic nanoroughness modulates neuron-astrocyte interactions and function via mechanosensing cation channels. Proc. Natl. Am. Sci. 111:16124–16129, 2014.

    Article  Google Scholar 

  5. Chapman, C. A. R., H. Chen, M. Stamou, J. Biener, M. M. Biener, P. J. Lein, and E. Seker. Nanoporous gold as a neural interface coating: effects of topography, surface chemistry, and feature size. ACS Appl. Mater. Interfaces 7:7093–7100, 2015.

    Article  Google Scholar 

  6. Chapman, C. A. R., P. Daggumati, S. C. Gott, M. P. Rao, and E. Seker. Substrate toporgraphy guides pore morphology evolution in nanoporous gold thin films. Scripta Mater. 110:33–36, 2016.

    Article  Google Scholar 

  7. Daggumati, P., Z. Matharu, L. Wang, and E. Seker. Biofouling-resilient nanoporous gold electrodes for DNA sensing. Anal. Chem. 87:8618–8622, 2015.

    Article  Google Scholar 

  8. Dalby, M. J., N. Gadegaard, and R. O. C. Oreffo. Harnessing nanotopography and integrin-matric interactions to influence stem cell fate. Nat. Mater. 13:558–569, 2014.

    Article  Google Scholar 

  9. Dembo, M., D. Torney, K. Saxman, and D. Hammer. The reaction-limited kinetics of membrane-to-surface adhesion and detachment. Proc. R. Soc. Lond. Ser. B: Biol. Sci. 234:55–83, 1988.

    Article  Google Scholar 

  10. Dorofeeva, T. S., and E. Seker. Electrically-tunable pore morphology in nanoporous gold thin films. Nano Res. 8(7):2188–2198, 2015.

    Article  Google Scholar 

  11. Erlebacher, J., M. J. Aziz, A. Karma, N. Dimitrov, and K. Sieradzki. Evolution of nanoporosity in dealloying. Nature 410:450–453, 2001.

    Article  Google Scholar 

  12. Feng, J., W. Zhao, and J. Wu. A label-free optical sensor based on nanoporous gold arrays for the detection of oligodeoxynucleotides. Biosens. Bioelectron. 30:21–27, 2011.

    Article  Google Scholar 

  13. Gasiorowski, J. Z., S. J. Liliensiek, P. Russel, D. A. Stephan, P. A. Nealey, and C. J. Murphy. Alterations in gene expression of human vascular endothelial cells associated with nanotopographic cues. Biomaterials 31:8882–8888, 2010.

    Article  Google Scholar 

  14. Geiger, B., J. P. Spatz, and A. D. Bershadsky. Environmental sensing through focal adhesions. Nat. Rev. Mol. Cell Bio. 10:21–33, 2009.

    Article  Google Scholar 

  15. Goldberg, M., R. Langer, and X. Jia. Nanostructured materials for applications in drug delivery and tissue engineering. J. Biomater. Sci. Polym. Ed. 18:241–268, 2007.

    Article  Google Scholar 

  16. Gultepe, E., D. Nagesha, S. Sridhar, and M. Amiji. Nanoporous inorganic membranes or coatings for sustained drug delivery in implantable devices. Adv. Drug Del. Rev. 62:305–315, 2010.

    Article  Google Scholar 

  17. Jeon, H., S. Koo, W. M. Reese, P. Loskill, C. P. Grigoropoulos, and K. E. Healy. Directing cell migration and organization via nanocrater-patterned cell-repellent interfaces. Nat. Mater. 14:918–923, 2015.

    Article  Google Scholar 

  18. Jin, H. J., and J. Weissmuller. A material with electrically tunable strength and flow stress. Science 332:1179–1182, 2011.

    Article  Google Scholar 

  19. Kang, K., S. Lee, J. E. Han, J. W. Choi, and M. Song. The complex morphology of reactive astrocytes controlled by fibroblast growth factor signaling. Glia 62:1328–1344, 2014.

    Article  Google Scholar 

  20. Kotov, N. A., J. O. Winter, I. P. Clements, E. Jan, B. P. Timko, S. Campidelli, S. Pathak, A. Mazzatenta, C. M. Lieber, M. Prato, R. V. Bellamkonda, G. A. Silva, N. W. S. Kam, F. Patolsky, and L. Ballerini. Nanomaterials for neural interfaces. Adv. Mater. 21:3970–4004, 2009.

    Article  Google Scholar 

  21. Kurtulus, O., P. Daggumati, and E. Seker. Molecular release from patterned nanoporous gold thin films. Nanoscale 6:7062–7071, 2014.

    Article  Google Scholar 

  22. Li, K., J. Huang, G. Shi, W. Zhang, and L. Jin. A sensitive nanoporous gold-based electrochemical DNA biosensor for Escherichia coli detection. Anal. Lett. 44:2559–2570, 2011.

    Article  Google Scholar 

  23. Mody, N. A., and M. R. King. Influence of Brownian motion on blood platelet flow behavior and adhesive dynamics near a planar wall. Langmuir 23:6321–6328, 2007.

    Article  Google Scholar 

  24. Morgan, J. T., C. J. Murphy, and P. Russell. What do mechanotransduction, Hippo, Wnt, and TGFB have in common? YAP and TAZ as key orchestrating molecules in ocular health and disease. Exp. Eye Res. 115:1–12, 2013.

    Article  Google Scholar 

  25. Patel, J., L. Radhakrishnan, B. Zhao, B. Uppalapati, R. C. Daniels, K. R. Ward, and M. M. Collinson. Electrochemical properties of nanostructured porous gold electrodes in biofouling solutions. Anal. Chem. 85:11610–11618, 2013.

    Article  Google Scholar 

  26. Polikov, V. S., P. A. Tresco, and W. M. Reichert. Response of brain tissue to chronically implanted neural electrodes. J. Neurosci. Methods 148:1–18, 2005.

    Article  Google Scholar 

  27. Popat, K. C., L. Leoni, C. A. Grimes, and T. A. Desai. Influence of engineered titania nanotubular surfaces on bone cells. Biomaterials 28:3188–3197, 2007.

    Article  Google Scholar 

  28. Robinson, D. A. The electrical properties of metal mircoelectrodes. Proc. IEEE 56:1065–1071, 1968.

    Article  Google Scholar 

  29. Santos, G. M., F. Zhao, J. Zeng, and W. Shih. Characterization of nanoporous gold disks for photothermal light harvesting and light-gated molecular release. Nanoscale 6:5718–5724, 2014.

    Article  Google Scholar 

  30. Seker, E., Y. Berdichevsky, M. R. Begley, M. L. Reed, K. J. Staley, and M. L. Yarmush. The fabrication of low-impedance nanoporous gold multiple-electrode arrays for neural electrophysiology studies. Nanotechnology 21:1–7, 2010.

    Article  Google Scholar 

  31. Seker, E., M. L. Reed, and M. R. Begley. Nanoporous gold: fabrication, characterization, and applications. Materials 2:2188–2215, 2009.

    Article  Google Scholar 

  32. Sofroniew, M. V. V., and H. V. Vinters. Astrocytes: biology and pathology. Acta Neuropathol. 119:7–35, 2010.

    Article  Google Scholar 

  33. Solanki, P. R., A. Kaushik, V. V. Agrawal, and B. D. Malhotra. Nanostructured metal oxide-based biosensors. NPG Asia Mater. 3:17–24, 2010.

    Article  Google Scholar 

  34. Venkatesh, H. S., T. B. Johung, V. Caretti, A. Noll, Y. Tang, S. Nagaraja, E. M. Gibson, C. W. Mount, J. Polepalli, S. S. Mitra, P. J. Woo, R. C. Malenka, H. Vogel, M. Bredel, P. Mallick, and M. Monje. Neuronal activity promotes glioma growth through neuroligin-3 secretion. Cell 161:803–816, 2015.

    Article  Google Scholar 

  35. Wagoner, N. J., J. Oh, P. Repovic, and E. N. Benveniste. Interleukin-6 production by astrocytes: autocrine regulation by IL-6 and the soluble IL-6 receptor. J. Neurosci. 19:5236–5244, 1999.

    Google Scholar 

  36. Woolley, A. J., H. A. Desai, and K. J. Otto. Chronic intracortical microelectrode arrays induce non-uniform, depth-related tissue responses. J. Neural Eng. 10:1–11, 2013.

    Article  Google Scholar 

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Acknowledgments

We gratefully acknowledge support from UC Lab Fees Research Program Award [12-LR-237197], Research Investments in the Sciences & Engineering (RISE) Award, and National Science Foundation Awards (CBET-1512745 and CBET&DMR-1454426). C. Chapman was supported by a National Science Foundation Graduate Research Fellowship [DGE-1148897] and a predoctoral fellowship from the National Institute of Health [T32-GM008799]. Support was also provided by the CounterACT Program, National Institutes of Health Office of the Director, and the National Institutes of Neurological Disorders and Stroke [U54-NS079202]. H. Chen was supported by a predoctoral fellowship from the National Institute of Environmental Health Sciences [T32-ES007059]. H. Chen and M. Stamou received predoctoral fellowships from the UC Davis Superfund Basic Research Program [P42-ES04699].

Conflict of Interest

E Seker has an issued patent, titled “Nanoporous metal multiple electrode array and method of making same”. C. A. R. Chapman, H. Chen, M. Stamou, and P. J. Lein declare that they have no conflicts of interest.

Ethical Standards

All studies were conducted according to protocols approved by the Institutional Animal Care and Use Committee of the University of California, Davis. No human subjects research was performed in this study.

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Author notes

  1. Marianna Stamou

    Present address: Department of Health Sciences and Technology, Swiss Federal Institute of Technology (ETH) Zurich, Zurich, Switzerland

Authors and Affiliations

  1. Department of Biomedical Engineering, University of California - Davis, Davis, CA, 95616, USA

    Christopher A. R. Chapman

  2. Department of Molecular Biosciences, University of California - Davis, Davis, CA, 95616, USA

    Hao Chen, Marianna Stamou & Pamela J. Lein

  3. Department of Electrical and Computer Engineering, University of California - Davis, 3177 Kemper Hall, Davis, CA, 95616, USA

    Erkin Seker

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  1. Christopher A. R. Chapman
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Correspondence to Erkin Seker.

Additional information

Associate Editor Tejal Desai oversaw the review of this article.

Dr. Erkin Şeker joined the Department of Electrical and Computer Engineering at UC Davis in 2011. He received his PhD degree in Electrical Engineering from the University of Virginia (UVA) in 2007, where he developed techniques to control mechanical and morphological properties of nanoporous gold. During his postdoctoral appointment in the Department of Chemistry at UVA he investigated material-biomolecule interactions and developed microfluidic flow control schemes. Between 2009 and 2011, as a research associate at the Center for Engineering in Medicine at Harvard Medical School, he developed multiple electrode arrays for neural electrophysiology applications and spearheaded the development of microsystems for monitoring transcriptional and secretory dynamics at a cellular-level in the context of metabolic dysregulation. At UC Davis, he is leading the interdisciplinary Multifunctional Nanoporous Metals research group with the overarching goal of understanding and controlling nanostructured material properties and their interaction with biological systems to develop effective biomedical tools for both basic and clinical applications. He is the recipient of Fund for Medical Discovery Award from Massachusetts General Hospital and a NSF CAREER Award. He served as an Associate Scientific Advisor for Science Translational Medicine journal and was selected to participate in National Academy of Engineering’s Annual Frontiers of Engineering Education Symposium in 2014.

This article is part of the 2016 Young Innovators Issue.

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Chapman, C.A.R., Chen, H., Stamou, M. et al. Mechanisms of Reduced Astrocyte Surface Coverage in Cortical Neuron-Glia Co-cultures on Nanoporous Gold Surfaces. Cel. Mol. Bioeng. 9, 433–442 (2016). https://doi.org/10.1007/s12195-016-0449-4

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