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Biofunctional polymer nanoparticles for intra-articular targeting and retention in cartilage

Nature Materials volume 7, pages 248–254 (2008)Cite this article

Abstract

The extracellular matrix of dense, avascular tissues presents a barrier to entry for polymer-based therapeutics, such as drugs encapsulated within polymeric particles. Here, we present an approach by which polymer nanoparticles, sufficiently small to enter the matrix of the targeted tissue, here articular cartilage, are further modified with a biomolecular ligand for matrix binding. This combination of ultrasmall size and biomolecular binding converts the matrix from a barrier into a reservoir, resisting rapid release of the nanoparticles and clearance from the tissue site. Phage display of a peptide library was used to discover appropriate targeting ligands by biopanning on denuded cartilage. The ligand WYRGRL was selected in 94 of 96 clones sequenced after five rounds of biopanning and was demonstrated to bind to collagen II α1. Peptide-functionalized nanoparticles targeted articular cartilage up to 72-fold more than nanoparticles displaying a scrambled peptide sequence following intra-articular injection in the mouse.

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Figure 1: Phage amplification and competitive binding showing that C1–3 was selected owing to its specificity of binding to the cartilage matrix.
Figure 2: Competitive and sequence-specific binding of WYRGRL.
Figure 3: Characterization of peptide-conjugated nanoparticles.
Figure 4: In vivo targeting of WYRGRL–PPS nanoparticles.
Figure 5: Quantification of nanoparticles in the ECM and within the cells.
Figure 6: Size-dependent matrix accumulation of WYRGRL–PPS after 24 h.

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Acknowledgements

The authors thank V. Garea and M. Pasquier for the help with the animal work and histology, A. Rehor for help with and discussions on initial nanoparticle synthesis, R. Schoenmakers for peptide synthesis, A. van der Vlies for obtaining 1H-NMR spectra, and especially the BIOP core facility for their support in establishing the imaging and quantification protocol and the proteomics core facility for running mass spectrometry analyses, and M. Lütolf for his help and suggestions in preparing the manuscript. DAR was supported by a grant from the Hans-Neuenschwander-Stiftung, Berne.

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Authors and Affiliations

  1. Institute of Bioengineering and Institute of Chemical Sciences and Engineering, Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland

    Dominique A. Rothenfluh, Harry Bermudez, Conlin P. O’Neil & Jeffrey A. Hubbell

  2. Department of Polymer Science and Engineering, University of Massachusetts, Amherst, Massachusetts 01003, USA

    Harry Bermudez

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  1. Dominique A. Rothenfluh
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Contributions

The experiments have been designed by D.A.R. and J.A.H. and carried out by D.A.R. H.B. introduced phage display and aided in establishing initial protocols. C.P.O. assisted in the affinity purification. The manuscript was written by D.A.R. and J.A.H. Principal investigator is J.A.H.

Corresponding author

Correspondence to Jeffrey A. Hubbell.

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The EPFL has filed an application for a patent on the technology described in this manuscript, of which D.A.R. and J.A.H. are inventors.

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Supplementary Information, Table S1

Supplementary Table S1 and Supplementary figures S1-S3 (PDF 343 kb)

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Rothenfluh, D., Bermudez, H., O’Neil, C. et al. Biofunctional polymer nanoparticles for intra-articular targeting and retention in cartilage. Nature Mater 7, 248–254 (2008). https://doi.org/10.1038/nmat2116

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