Administration of endothelial progenitor cells accelerates the resolution of arterial thrombus in mice

Giane D Carneiro; Micheli S Sielski; Cristiano Pedroso Vieira; Fabio Trindade Maranhão Costa; Claudio C Werneck; Cristina P Vicente

doi:10.1016/j.jcyt.2019.01.005

Administration of endothelial progenitor cells accelerates the resolution of arterial thrombus in mice

Cytotherapy. 2019 Apr;21(4):444-459. doi: 10.1016/j.jcyt.2019.01.005. Epub 2019 Mar 20.

Authors

Giane D Carneiro¹, Micheli S Sielski¹, Cristiano Pedroso Vieira², Fabio Trindade Maranhão Costa³, Claudio C Werneck⁴, Cristina P Vicente⁵

Affiliations

¹ Structural and Functional Biology Department, State University of Campinas (UNICAMP), São Paulo, Brazil.
² Pharmacology Department, State University of Campinas (UNICAMP), São Paulo, Brazil.
³ Genetics, Evolution and Bioagents Department, State University of Campinas (UNICAMP), Campinas, Brazil.
⁴ Biochemistry and Tissue Biology Department, Institute of Biology, State University of Campinas (UNICAMP), SãoPaulo, Brazil.
⁵ Structural and Functional Biology Department, State University of Campinas (UNICAMP), São Paulo, Brazil. Electronic address: [email protected].

PMID: 30904331
DOI: 10.1016/j.jcyt.2019.01.005

Abstract

Background: Endothelial progenitor cells (EPCs) are circulating progenitor cells that can play an essential role in vascular remodelling. In this work, we compared the role of two EPCs cultivated with different mediums in the resolution of the arterial thrombus induced by FeCl₃ lesion and in vessel re-endothelization in the mouse carotid artery.

Methods: Mice mononuclear cells were differentiated into EPCs using Dulbecco's Modified Eagle's Medium (DMEM) and vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF) and IGF (Insulin Growth Factor) called EPCs--M1) or with EGM2(endothelial growth medium) (media supplemented with growth factors from Lonza called (EPCs-M2) for 30days and characterized using flow cytometry. The animals received three EPC injections post-lesion, and we analyzed thrombosis time, vessel re-endothelization, metalloproteinases activities, eNOS (endothelial Nitric oxide synthase) presence and SDF-1(Stromal Derived Factor- 1) levels in circulation.

Results: EPC-M1 presented a more immature progenitor profile than EPC-M2 cells. The injection of EPC-M1 prolonged the thrombosis time, and the treatment with the different EPCs increased eNOS expression and MMP2 (Metalloproteinase 2) activity and decreased SDF-1 in plasma. Only EPC-M1 treatment increased both MMP2 and MMP9 and reduced thrombus after 7days. Also, both EPCs decreased platelet aggregation in vitro.

Conclusions: EPCs-M1 were more efficient in all of the analyzed assays. EPCsM2 may be a more mature EPC, proliferating less and promoting a less significant matrix remodelling. EPCs can promote vascular remodelling by inhibiting thrombosis and stimulating vascular wall remodelling and the treatment with a more immature progenitor may be more efficient in this process.

Keywords: MMPs (metalloproteinases); arterial thrombosis; eNOS (endothelial Nitric Oxide Synthase); endothelial progenitor cells; intravital microscopy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Arteries / pathology
Cell Differentiation
Cells, Cultured
Chemokine CXCL12 / metabolism
Embolization, Therapeutic
Endothelial Progenitor Cells / metabolism
Endothelial Progenitor Cells / transplantation*
Gelatinases / metabolism
Male
Matrix Metalloproteinase 2 / metabolism
Matrix Metalloproteinase 9 / metabolism
Mice, Inbred C57BL
Nitric Oxide Synthase Type III / metabolism
Platelet Aggregation
Thrombosis / enzymology
Thrombosis / pathology
Thrombosis / therapy*
Vascular Endothelial Growth Factor A / metabolism

Substances

Chemokine CXCL12
Vascular Endothelial Growth Factor A
Nitric Oxide Synthase Type III
Gelatinases
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9