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Direct Quantification of Solute Diffusivity in Agarose and Articular Cartilage Using Correlation Spectroscopy

Abstract

Articular cartilage is an avascular tissue; diffusive transport is critical for its homeostasis. While numerous techniques have been used to quantify diffusivity within porous, hydrated tissues and tissue engineered constructs, these techniques have suffered from issues regarding invasiveness and spatial resolution. In the present study, we implemented and compared two separate correlation spectroscopy techniques, fluorescence correlation spectroscopy (FCS) and raster image correlation spectroscopy (RICS), for the direct, and minimally-invasive quantification of fluorescent solute diffusion in agarose and articular cartilage. Specifically, we quantified the diffusional properties of fluorescein and Alexa Fluor 488-conjugated dextrans (3k and 10k) in aqueous solutions, agarose gels of varying concentration (i.e. 1, 3, 5%), and in different zones of juvenile bovine articular cartilage explants (i.e. superficial, middle, and deep). In agarose, properties of solute diffusion obtained via FCS and RICS were inversely related to molecule size, gel concentration, and applied strain. In cartilage, the diffusional properties of solutes were similarly dependent upon solute size, cartilage zone, and compressive strain; findings that agree with work utilizing other quantification techniques. In conclusion, this study established the utility of FCS and RICS as simple and minimally invasive techniques for quantifying microscale solute diffusivity within agarose constructs and articular cartilage explants.

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Similar content being viewed by others Explore related subjectsDiscover the latest articles and news from researchers in related subjects, suggested using machine learning. Abbreviations
10k dextran:

AlexaFluor® 488-conjugated 10 kilodalton dextran

3k dextran:

AlexaFluor® 488-conjugated 3 kilodalton dextran

α :

Anomalous diffusion exponent

D :

Diffusion coefficient (time invariant parameter)

FCS:

Fluorescence correlation spectroscopy

FRAP:

Fluorescence recovery after photobleaching

Γ :

Transport coefficient (time invariant parameter)

ICS:

Image correlation spectroscopy

LSCM:

Laser scanning confocal microscopy

MRI:

Magnetic resonance imaging

NMR:

Nuclear magnetic resonance

PSF:

Point spread function

RICS:

Raster image correlation spectroscopy

ROI:

Region of interest

ω 0 :

Radial waist dimension of the microscope point spread function

ω z :

Axial waist dimension of the microscope point spread function

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Acknowledgments

The research reported herein was supported by the NIH under Award Number R21AR062738. Microscopy equipment was acquired through NIH shared instrumentation grants (S10 RR0272773 & S10 OD016361) and access was supported by NIGMS (P20 GM103446). The content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. We thank Dr. Jeffrey Caplan and Michael Moore in the Delaware Biotechnology Institute Bio-Imaging Center for their technical assistance, as well as Ms. Alison Wright for assisting in the analysis of our correlation spectroscopy data.

Conflict of interest

The authors report no conflicts of interest with the submitted work.

Author information Authors and Affiliations

  1. Biomechanics & Movement Science, University of Delaware, Newark, DE, USA

    Janty S. Shoga & Christopher Price

  2. Department of Mechanical Engineering, University of Delaware, Newark, DE, USA

    Brian T. Graham, Liyun Wang & Christopher Price

  3. Department of Biomedical Engineering, University of Delaware, 161 Colburn Lab, Newark, DE, 19716, USA

    Christopher Price

Authors
  1. Janty S. Shoga
  2. Brian T. Graham
  3. Liyun Wang
  4. Christopher Price
Contributions

All authors have read and approved the final submission of the manuscript. The contributions of the authors are described below: Study Design – JS, CP. Acquisition of Data – JS, BG. Analysis and Interpretation – JS, BG, LW, CP. Manuscript Preparation – JS, BG, LW, CP. Statistical Analysis – JS, CP.

Corresponding author

Correspondence to Christopher Price.

Additional information

Associate Editor Michael S. Detamore oversaw the review of this article.

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Shoga, J.S., Graham, B.T., Wang, L. et al. Direct Quantification of Solute Diffusivity in Agarose and Articular Cartilage Using Correlation Spectroscopy. Ann Biomed Eng 45, 2461–2474 (2017). https://doi.org/10.1007/s10439-017-1869-6

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