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In Situ Quantification of Surface Chemistry in Porous Collagen Biomaterials

  • Nondestructive Characterization of Biomaterials for Tissue Engineering and Drug Delivery
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Abstract

Cells inside a 3D matrix (such as tissue extracellular matrix or biomaterials) sense their insoluble environment through specific binding interactions between their adhesion receptors and ligands present on the matrix surface. Despite the critical role of the insoluble matrix in cell regulation, there exist no widely-applicable methods for quantifying the chemical stimuli provided by a matrix to cells. Here, we describe a general-purpose technique for quantifying in situ the density of ligands for specific cell adhesion receptors of interest on the surface of a 3D matrix. This paper improves significantly the accuracy of the procedure introduced in a previous publication by detailed marker characterization, optimized staining, and improved data interpretation. The optimized methodology is utilized to quantify the ligands of integrins α 1 β 1, α 2 β 1 on two kinds of matched porous collagen scaffolds, which are shown to possess significantly different ligand density, and significantly different ability to induce peripheral nerve regeneration in vivo. Data support the hypothesis that cell adhesion regulates contractile cell phenotypes, recently shown to be inversely related to organ regeneration. The technique provides a standardized way to quantify the surface chemistry of 3D matrices, and a means for introducing matrix effects in quantitative biological models.

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Acknowledgments

The cDNA of I domains (α1, α2) of integrin subunits α1 and α2 were a kind gift of Sue Craig (Martin Humphries Lab, University of Manchester, UK). We thank Dr. Amit Roy for performing the initial protein expressions and purifications, Mrs Debby Pheasant (MIT biophysical instrumentation facility) for valuable technical assistance in the circular dichroism experiments, Dr. Albert Tai (genomics facility core, Tufts University) for valuable technical assistance in the BIACORE experiments, and Prof. K. Van Vliet for providing access to the fluorescent microscope of her lab. The authors acknowledge financial support from the National Institutes of Health grant RO1 NS051320 (I.V.Y., D.S.T, E.C.S., P.T.C.S), the Biomechanics Training Grant T32EB006348 (E.C.S.), the National Science Foundation Graduate Research Fellowship DGE‐1122374 (M.C.B), Singapore Alliance for Science and Technology Center, BioSym IRG and Singapore and MIT Alliance 2, CSB program (P.T.C.S, and D.S.T.), and National Institutes of Health grant 9P41EB015871-26A1 (P.T.C.S).

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Authors and Affiliations

  1. Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA

    Dimitrios S. Tzeranis, Eric C. Soller, Melissa C. Buydash, Peter T. C. So & Ioannis V. Yannas

  2. Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA

    Peter T. C. So & Ioannis V. Yannas

  3. Department of Mechanical Engineering, National Technical University of Athens, 15780, Zografou, Greece

    Dimitrios S. Tzeranis

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  1. Dimitrios S. Tzeranis
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Correspondence to Dimitrios S. Tzeranis.

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Associate Editor Agata A. Exner oversaw the review of this article.

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Tzeranis, D.S., Soller, E.C., Buydash, M.C. et al. In Situ Quantification of Surface Chemistry in Porous Collagen Biomaterials. Ann Biomed Eng 44, 803–815 (2016). https://doi.org/10.1007/s10439-015-1445-x

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  • DOI: https://doi.org/10.1007/s10439-015-1445-x

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