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Elucidation of Endothelial Cell Hemostatic Regulation with Integrin-Targeting Hydrogels

  • Allison Post
  • Sevinj Isgandarova
  • Margarita Martinez-Moczygemba
  • Mariah Hahn
  • Brooke Russell
  • Magnus Hook
  • Elizabeth Cosgriff-HernandezEmail author
Article

Abstract

Despite advances in the development of materials for cardiovascular devices, current strategies generally lack the thromboresistance of the native endothelium both in terms of efficacy and longevity. To harness this innate hemostatic regulation and improve long-term hemocompatibility, biohybrid devices are designed to promote endothelialization. Much of the research effort to date has focused on the use of extracellular matrix (ECM)-mimics and coatings to promote endothelial cell adhesion and migration with less attention given to the effect of the supported ECM binding events on hemostatic regulation. In this study, we developed integrin-targeted hydrogels to investigate the individual and combined effects of integrin binding events supported by many ECM-based coatings (α1β1, α2β1, α5β1, αvβ3). Targeted endothelial cell integrin interactions were first confirmed with antibody blocking studies and then correlated with gene expression of hemostatic regulators and a functional assay of platelet attachment and activation. Surfaces that targeted integrins α1β1 and α2β1 resulted in an endothelial cell layer that exhibited a thromboresistant phenotype with an associated reduction in platelet attachment and activation. It is anticipated that identification of specific integrins that promote endothelial cell adhesion as well as thromboresistance will enable the design of cardiovascular materials with improved long-term hemocompatibility.

Keywords

Endothelial cells Integrins Hemostasis Thromboresistance Platelets 

Notes

Acknowledgments

This work was funded by the National Institutes of Health, Grant Numbers R21 EB020978 and R01 EB013297. The authors kindly thank the Grande-Allen lab for their help in platelet adhesion studies.

Conflict of interest

Dr. Cosgriff-Hernandez reports a stakeholder interest in ECM Technologies, outside the submitted work; In addition, Dr. Cosgriff-Hernandez, Dr. Magnus Hook, and Dr. Brooke Russell have a patent US20110288274A1 issued, and a patent US9725498B2 issued. Dr. Hook and Dr. Russell are also stakeholders in ECM Technologies (ECMT) that seeks to commercialize designer collagens such as those used in the studies presented.

Supplementary material

10439_2018_2194_MOESM1_ESM.tif (11.4 mb)
Supplementary Fig. 1 A) Cell population selected for analysis in the SSC vs. FSC plot. B) Representative PE signal from FACS analysis. The dotted line represents a blank sample and the grey peak is the signal from the stained HUVECs. Supplementary material 1 (TIFF 11648 kb)
10439_2018_2194_MOESM2_ESM.tif (22.1 mb)
Supplementary Fig. 2 Representative images of unblocked and most blocked (blocking with integrin antibody combinations) HUVEC adhesion. Nucleic stain: SYBR Green; actin stain: rhodamine phalloidin (red). Scale bar represents 200 µm. Supplementary material 2 (TIFF 22627 kb)
10439_2018_2194_MOESM3_ESM.tif (9.4 mb)
Supplementary Fig. 3 HUVEC gene expression of hemostatic regulators. All values have been normalized to TCPS GAPDH (value of 1, no fold change). Data shown is average ± standard deviation. * represents significance from all other groups. Significance was determined by ANOVA with a post hoc Tukey’s test with p < 0.05 for all comparisons. Supplementary material 3 (TIFF 9606 kb)
10439_2018_2194_MOESM4_ESM.docx (13 kb)
Supplementary material 4 (DOCX 13 kb)

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Copyright information

© Biomedical Engineering Society 2019

Authors and Affiliations

  1. 1.Department of Biomedical EngineeringTexas A&M UniversityCollege StationUSA
  2. 2.Center for Inflammatory and Infectious Diseases, Institute of Biosciences and TechnologyTexas A&M Health Science CenterHoustonUSA
  3. 3.Department of Biomedical EngineeringRensselaer Polytechnic InstituteTroyUSA
  4. 4.Department of Biomedical EngineeringUniversity of TexasAustinUSA

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