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Estimating Cell Concentration in Three-Dimensional Engineered Tissues Using High Frequency Quantitative Ultrasound

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Abstract

Histology and biochemical assays are standard techniques for estimating cell concentration in engineered tissues. However, these techniques are destructive and cannot be used for longitudinal monitoring of engineered tissues during fabrication processes. The goal of this study was to develop high-frequency quantitative ultrasound techniques to nondestructively estimate cell concentration in three-dimensional (3-D) engineered tissue constructs. High-frequency ultrasound backscatter measurements were obtained from cell-embedded, 3-D agarose hydrogels. Two broadband single-element transducers (center frequencies of 30 and 38 MHz) were employed over the frequency range of 13–47 MHz. Agarose gels with cell concentrations ranging from 1 × 104 to 1 × 106 cells mL−1 were investigated. The integrated backscatter coefficient (IBC), a quantitative ultrasound spectral parameter, was calculated and used to estimate cell concentration. Accuracy and precision of this technique were analyzed by calculating the percent error and coefficient of variation of cell concentration estimates. The IBC increased linearly with increasing cell concentration. Axial and lateral dimensions of regions of interest that resulted in errors of less than 20% were determined. Images of cell concentration estimates were employed to visualize quantitatively regional differences in cell concentrations. This ultrasound technique provides the capability to rapidly quantify cell concentration within 3-D tissue constructs noninvasively and nondestructively.

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Acknowledgments

The authors thank Sally Z. Child, M.S., and Stephen McAleavey, Ph.D. (University of Rochester) for technical assistance and helpful discussions. This work was supported, in part, by National Institutes of Health Grant EB008368.

Conflict of interest

No conflicts of interest exist.

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Authors and Affiliations

  1. Department of Biomedical Engineering, University of Rochester, 310 Goergen Hall, P.O. Box 270168, Rochester, NY, 14627, USA

    Karla P. Mercado, Denise C. Hocking & Diane Dalecki

  2. The Rochester Center for Biomedical Ultrasound, University of Rochester, Rochester, NY, USA

    Karla P. Mercado, María Helguera, Denise C. Hocking & Diane Dalecki

  3. Chester F. Carlson Center for Imaging Science, Rochester Institute of Technology, Rochester, NY, USA

    María Helguera

  4. Department of Pharmacology and Physiology, University of Rochester, Rochester, NY, USA

    Denise C. Hocking

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  1. Karla P. Mercado
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Correspondence to Diane Dalecki.

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Associate Editor Scott I Simon oversaw the review of this article.

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Mercado, K.P., Helguera, M., Hocking, D.C. et al. Estimating Cell Concentration in Three-Dimensional Engineered Tissues Using High Frequency Quantitative Ultrasound. Ann Biomed Eng 42, 1292–1304 (2014). https://doi.org/10.1007/s10439-014-0994-8

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  • DOI: https://doi.org/10.1007/s10439-014-0994-8

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