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Multiphoton microscopy guides neurotrophin modification with poly(ethylene glycol) to enhance interstitial diffusion

Nature Materials volume 3, pages 489–494 (2004)Cite this article

Abstract

Brain-derived neurotrophic factor (BDNF) is a promising therapeutic agent for the treatment of neurodegenerative diseases. However, the limited distribution of this molecule after administration into the brain tissue considerably hampers its efficacy. Here, we show how multiphoton microscopy of fluorescently tagged BDNF in brain-tissue slices provides a useful and rapid screening method for examining the diffusion of large molecules in tissues, and for studying the effects of chemical modifications—for example, conjugating with polyethylene glycol (PEG)—on the diffusion constant. This single variable, obtained by monitoring short-term diffusion in real time, can be effectively used for rational drug design. In this study on fluorescently tagged BDNF and BDNF-PEG, we identify slow diffusion as a major contributing factor to the limited penetration of BDNF, and demonstrate how chemical modification can be used to overcome this barrier.

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Figure 1: MALDI-TOF spectra for native BDNF and BDNF-PEG conjugate.
Figure 2
Figure 3: Intensity profile data and model fits initially (solid squares and lines, respectively) and at later times (open squares and dashed lines, respectively).
Figure 4: MPFPR recovery trace for BNDF-R.
Figure 5: Simulated tissue neurotrophin concentration in the vicinity of a controlled-release pellet.

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Acknowledgements

We thank Moses V. Chao for his generous gift of the trkB-expressing PC12 cell line, and Amgen for the kind donation of all BDNF used in this study. This work was supported by grants from the National Institutes of Health (NS-38470 and NS-45236, S.C.M., M.S. and W.M.S.). This publication was made possible in part by Grant Number P41-RR04224 from the National Center for Research Resources, National Institutes of Health to R.M.W, W.R.Z. and W.W.W.

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Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, Cornell University, Ithaca, 14853, New York, USA

    Mark Stroh & W. Mark Saltzman

  2. Applied and Engineering Physics, Cornell University, Ithaca, 14853, New York, USA

    Warren R. Zipfel, Rebecca M. Williams & Watt W. Webb

  3. Department of Biomedical Engineering, Yale university, New Haven, 06520, Connecticut, USA

    Shu Chin Ma & W. Mark Saltzman

  4. Department of Biomedical Engineering, Yale University, PO Box 208284, New Haven, 06520, Connecticut, USA

    W. Mark Saltzman

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Correspondence to W. Mark Saltzman.

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Stroh, M., Zipfel, W., Williams, R. et al. Multiphoton microscopy guides neurotrophin modification with poly(ethylene glycol) to enhance interstitial diffusion. Nature Mater 3, 489–494 (2004). https://doi.org/10.1038/nmat1159

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