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Biomechanical regulation of blood vessel growth during tissue vascularization

Nature Medicine volume 15, pages 657–664 (2009)Cite this article

Abstract

Formation of new vessels in granulation tissue during wound healing has been assumed to occur solely through sprouting angiogenesis. In contrast, we show here that neovascularization can be accomplished by nonangiogenic expansion of preexisting vessels. Using neovascularization models based on the chick chorioallantoic membrane and the healing mouse cornea, we found that tissue tension generated by activated fibroblasts or myofibroblasts during wound contraction mediated and directed translocation of the vasculature. These mechanical forces pulled vessels from the preexisting vascular bed as vascular loops with functional circulation that expanded as an integral part of the growing granulation tissue through vessel enlargement and elongation. Blockade of vascular endothelial growth factor receptor-2 confirmed that biomechanical forces were sufficient to mediate the initial vascular growth independently of endothelial sprouting or proliferation. The neovascular network was further remodeled by splitting, sprouting and regression of individual vessels. This model explains the rapid appearance of large functional vessels in granulation tissue during wound healing.

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Figure 1: Vascularization of a fibrin and collagen matrix implanted on the CAM through elongation of the preexisting capillary network.
Figure 2: Neovascularization is preceded by ingrowth of proto- and myofibroblasts.
Figure 3: Matrix contraction is a prerequisite for vascular ingrowth on the CAM.
Figure 4: Time-lapse recordings of vessel remodeling during implant vascularization on the CAM.
Figure 5: Neovascularization of the wounded mouse cornea by recruitment of functional microvascular loops from the limbal capillary network.
Figure 6: Characterization of neovascular growth during wound healing of the injured mouse cornea using whole mount staining.

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Acknowledgements

We thank M. Aronsson for help with cornea experiments, J. Kilarska for help with image processing and analysis and K. Kullander and N. Rabe for help with microscopy of whole-mount corneas. We also thank M. Swartz for allowing us to use her laboratory to perform gel contraction and image analysis and A. Bikfalvi for comments. This study was supported by grants to P.G. from the Göran Gustafsson Foundation, the Swedish Cancer Foundation (project 4422-B04-O5XAB), the Swedish Research Council (project K2005-31X-15348-01A), the Children's Cancer Foundation of Sweden (project 04/037), Lions Cancer Research Foundation in Uppsala, the Magnus Bergvall Foundation, King Gustaf V's 80-Year Foundation and Uppsala Foundation for Medical Research, by grants to A.K. from the Crown Princess Margareta Foundation (KMA), the Edwin Jordan Foundation, Karolinska Institute research grants, Stiftelsen Synfrämjandets Forskningsfond, and Swedish Research Council (project 2006-13828-37891-37) and by grants to B.S. from St. Erik Eye Hospital Research Foundation, Sigvard and Marianne Bernadotte Research Foundation for Children's Eye Care and Stiftelsen Synfrämjandets Forskningsfond.

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Author notes

  1. Present address: Laboratory of Mechanobiology and Morphogenesis, Swiss Federal Institute of Technology, Lausanne, Switzerland.

  2. Branka Samolov and Ludvig Petersson: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Medical Biochemistry and Microbiology, Biomedical Center, University of Uppsala, Uppsala, Sweden

    Witold W Kilarski, Ludvig Petersson & Pär Gerwins

  2. Department of Clinical Neuroscience, Section of Ophthalmology and Vision, Karolinska Institutet, Stockholm, Sweden

    Branka Samolov & Anders Kvanta

  3. Department of Oncology, Radiology, and Clinical Immunology, Section for Radiology, Uppsala University Hospital, Uppsala, Sweden

    Pär Gerwins

Authors
  1. Witold W Kilarski
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  2. Branka Samolov
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  4. Anders Kvanta
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Contributions

W.W.K. was involved in the overall design of the study; performed most of the experiments involving the CAM, immunohistochemistry, in vitro gel contraction, whole-mount staining of cornea samples and statistical analysis; analyzed data; and wrote most of the manuscript. B.S. designed and performed the cornea experiments and statistical analysis, and she wrote portions of the manuscript and edited the manuscript. L.P. performed CAM experiments with PVA sponges, in vitro cell analysis, peptide inhibitor studies and cornea stainings. A.K. designed parts of cornea experiments and edited the manuscript. P.G. designed the overall study, analyzed data, and wrote and edited the manuscript.

Corresponding authors

Correspondence to Witold W Kilarski or Pär Gerwins.

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Kilarski, W., Samolov, B., Petersson, L. et al. Biomechanical regulation of blood vessel growth during tissue vascularization. Nat Med 15, 657–664 (2009). https://doi.org/10.1038/nm.1985

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