Type I Diabetes Delays Perfusion and Engraftment of 3D Constructs by Impinging on Angiogenesis; Which can be Rescued by Hepatocyte Growth Factor Supplementation



The biggest bottleneck for cell-based regenerative therapy is the lack of a functional vasculature to support the grafts. This problem is exacerbated in diabetic patients, where vessel growth is inhibited. To address this issue, we aim to identify the causes of poor vascularization in 3D engineered tissues in diabetes and to reverse its negative effects.


We used 3D vascularized constructs composed of microvessel fragments containing all cells present in the microcirculation, embedded in collagen type I hydrogels. Constructs were either cultured in vitro or implanted subcutaneously in non-diabetic or in a type I diabetic (streptozotocin-injected) mouse model. We used qPCR, ELISA, immunostaining, FACs and co-culture assays to characterize the effect of diabetes in engineered constructs.


We demonstrated in 3D vascularized constructs that perivascular cells secrete hepatocyte growth factor (HGF), driving microvessel sprouting. Blockage of HGF or HGF receptor signaling in 3D constructs prevented vessel sprouting. Moreover, HGF expression in 3D constructs in vivo is downregulated in diabetes; while no differences were found in HGF receptor, VEGF or VEGF receptor expression. Low HGF expression in diabetes delayed the inosculation of graft and host vessels, decreasing blood perfusion and preventing tissue engraftment. Supplementation of HGF in 3D constructs, restored vessel sprouting in a diabetic milieu.


We show for the first time that diabetes affects HGF secretion in microvessels, which in turn prevents the engraftment of engineered tissues. Exogenous supplementation of HGF, restores angiogenic growth in 3D constructs showing promise for application in cell-based regenerative therapies.

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Change history

  • 17 September 2019

    The values on the Y-axis for Figs. 1a, 2a, and 5a were in ng/L, not in molar. To meet the CAMB journal requirements, we have converted the values to molar and updated the graphs.


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This work was supported by grants from the Canadian Institutes of Health Research (CIHR), Institute of Circulatory and Respiratory Health (137352 and PJT153160) and the Heart and Stroke Foundation of Canada (G-14-0006265) to S.S.N; NIH Grant (EB007556) to J.B.H. A Discovery grant from the Natural Sciences and Engineering Research Council (RGPIN 06621-2017) and an Early Researcher Award from the Ministry of Research, Innovation and Science (ER17-13-149) to S.S.N. supported R.H.

Authors contribution

SSN designed the experiments, coordinated the project and contributed to the writing of the manuscript. JBH supported HGF blockage assays. WA designed and performed experiment and contributed to manuscript writing. RH contributed to performing experiments and writing manuscript and YA contributed to performing experiments.

Conflict of interest

J.B.H. is an inventor on a patent regarding the use of adipose-derived microvessels and has equity interest with Advanced Solutions Life sciences, which is commercializing isolated microvessel technology. A version of this technology was used as an experimental model in this manuscript. This equity was obtained after the work for this manuscript was completed. W.A., R.H., Y.A. and S.S.N. declare no conflict of interest.

Ethical standards

All animal studies were carried out in accordance with Institutional guidelines and approved by the Animal Care Committee at the University Health Network (ID 2420 and 2427). No human subjects were used in this study.

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Correspondence to Sara S. Nunes.

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Sara S. Nunes is a Scientist at the University Health Network in the Toronto General Hospital Research Institute. She holds an Assistant Professor appointment at the Institute of Biomaterials Biomedical Engineering and a cross-appointment at the Department of Laboratory Medicine & Pathobiology at the University of Toronto. Her translational research program aims to develop regenerative medicine strategies, and use bioengineering approaches to study cardiovascular diseases and for drug testing. Nunes obtained her Ph.D. from the State University of Rio de Janeiro, Brazil, and completed postdoctoral training under Dr. James Hoying, Ph.D. at the University of Louisville and later with Prof. Milica Radisic, Ph.D. at the University of Toronto. Dr. Nunes received several awards and fellowships for her work, including the prestigious Early Researcher Award from the Ministry of Research Innovation and Science in Canada and the Scientist Development Grant from the American Heart Association, USA. She has developed new vascularization techniques to support functional tissues for organ regeneration and is pioneering the work to create mature vessels with specific arterio-venous identities in 3D engineered tissues. Her work on human cardiac tissues-on-a-dish, named biowires, has opened a new area of research in human pluripotent stem cell-derived cardiomyocyte maturation and drug testing which catalyzed further mechanistic and translational research in this area worldwide. She holds funding from CIHR, NSERC, CFREF and JDRF-USA. She serves as a committee review member for Canadian Institutes of Health Research, and as Ad Hoc reviewer for NIH study section.


This article is part of the 2019 CMBE Young Innovators special issue.

Associate Editor Stephanie Michelle Willerth oversaw the review of this article.

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Altalhi, W., Hatkar, R., Hoying, J.B. et al. Type I Diabetes Delays Perfusion and Engraftment of 3D Constructs by Impinging on Angiogenesis; Which can be Rescued by Hepatocyte Growth Factor Supplementation. Cel. Mol. Bioeng. 12, 443–454 (2019). https://doi.org/10.1007/s12195-019-00574-3

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  • Endothelial cell
  • Tissue engineering
  • Microvessel
  • Regenerative medicine
  • Revascularization
  • Angiogenesis
  • Hepatocyte growth factor
  • Blood perfusion
  • Diabetes
  • Anastomosis
  • Inosculation