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In vivo laser speckle imaging reveals microvascular remodeling and hemodynamic changes during wound healing angiogenesis

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Abstract

Laser speckle contrast imaging (LSCI) is a high-resolution and high contrast optical imaging technique often used to characterize hemodynamic changes in short-term physiological experiments. In this study, we demonstrate the utility of LSCI for characterizing microvascular remodeling and hemodynamic changes during wound healing angiogenesis in vivo. A 2 mm diameter hole was made in the mouse ear and the periphery of the wound imaged in vivo using LSCI over 12 days. We were able to visualize and quantify the vascular and perfusion changes that accompanied wound healing in the microenvironment proximal to the wound, and validated these changes with histology. We found that consistent with the stages of wound healing, microvessel density increased during the initial inflammatory phase (i.e., day 0–3), stayed elevated through the tissue formation phase (i.e., until day 7) and returned to baseline during the tissue remodeling phase (i.e., by day 12). Concomitant “wide area mapping” of blood flow revealed that tissue perfusion in the wound periphery initially decreased, gradually increased from day 3–7, and subsided as healing completed. Interestingly, some regions exhibited a reestablishment of tissue perfusion approximately 6 days earlier than the ~18 days usually reported for the long term remodeling phase. The results from this study demonstrate that LSCI is an ideal platform for elucidating in vivo changes in microvascular hemodynamics and angiogenesis, and has the potential to offer invaluable insights in a range of disease models involving abnormal hemodynamics, such as diabetes and tumors.

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Acknowledgments

This work was supported jointly by National Institute of Aging award number R01AG029681, Department of Health and Human Services award number 1R43CA139983-01, and a JHU Institute of NanoBiotechnology (INBT) Junior Faculty Pilot Award. We gratefully acknowledge Peng Miao for assistance with image registration.

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Authors and Affiliations

  1. Department of Biomedical Engineering, The Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Abhishek Rege & Nitish V. Thakor

  2. Russell H. Morgan Department of Radiology and Radiological Sciences, The Johns Hopkins University School of Medicine, 720 Rutland Ave, 217 Traylor Bldg, Baltimore, MD, 21205, USA

    Kevin Rhie & Arvind P. Pathak

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  1. Abhishek Rege
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Correspondence to Arvind P. Pathak.

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Rege, A., Thakor, N.V., Rhie, K. et al. In vivo laser speckle imaging reveals microvascular remodeling and hemodynamic changes during wound healing angiogenesis. Angiogenesis 15, 87–98 (2012). https://doi.org/10.1007/s10456-011-9245-x

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