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Research article
Angiogenic properties of human endothelial colony-forming cells in Granulomatosis with Polyangiitis
Zoraida Sachetto
Universidade Estadual de Campinas
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
BACKGROUND
Endothelial progenitor cells are essential for vascular homeostasis. Considering the recurrent nature of granulomatosis with polyangiitis (GPA) the aim of the study was to evaluate the angiogenic capacity of endothelial colony-forming cells (ECFC), which have the capacity for neovasculogenesis in vitro, of the patients with GPA, before and after plasma stimulation.
METHODS
Thirteen GPA PR3-positive patients and 15 healthy controls were included. ECFC were isolated from periferic blood and characterized by flow cytometry (FACS). Capillary tube formation (Matrigel assay) and scratching assay were measured during 24 hours. The migration assay was also performed after overnight incubation with healthy control plasma and active GPA patient plasma. Three patients with active disease where submitted to recollection of ECFC after treatment for longitudinal evaluation.
RESULTS
The culture was successful in 62% of GPA patients and 57% of controls. In the matrigel assay the ECFC from patients and controls showed similar capillary structures formation, however ECFC from inactive GPA alone showed early loss of angiogenic capacity between 15 and 24 hours. In scratching assay, there was an impairment in the proliferative capacity of the ECFC between GPA patients and controls without significant difference (12th hour, p = 0.05). When incubated with control plasma, ECFC of remission GPA patients showed a significant lower migration capacity after the 4th hour (p = 0.0001). In longitudinal analysis, ECFC isolated after treatment showed significantly lower migration rates.
CONCLUSIONS
PR3-positive remission GPA ECFC demonstrated early loss of tube formation and less proliferative capacity compared to those of healthy controls, suggesting impairment of endothelial function.
Keywords
Anti-Neutrophil Cytoplasmic Antibody-Associated Vasculitis, Granulomatosis with Polyangiitis, Endothelial Progenitor Cells, Endothelial Colony-forming Cells
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This is a preprint. It has not completed peer review.
Research article
Angiogenic properties of human endothelial colony-forming cells in Granulomatosis with Polyangiitis
Zoraida Sachetto
Universidade Estadual de Campinas
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 22 May, 2020
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
BACKGROUND
Endothelial progenitor cells are essential for vascular homeostasis. Considering the recurrent nature of granulomatosis with polyangiitis (GPA) the aim of the study was to evaluate the angiogenic capacity of endothelial colony-forming cells (ECFC), which have the capacity for neovasculogenesis in vitro, of the patients with GPA, before and after plasma stimulation.
METHODS
Thirteen GPA PR3-positive patients and 15 healthy controls were included. ECFC were isolated from periferic blood and characterized by flow cytometry (FACS). Capillary tube formation (Matrigel assay) and scratching assay were measured during 24 hours. The migration assay was also performed after overnight incubation with healthy control plasma and active GPA patient plasma. Three patients with active disease where submitted to recollection of ECFC after treatment for longitudinal evaluation.
RESULTS
The culture was successful in 62% of GPA patients and 57% of controls. In the matrigel assay the ECFC from patients and controls showed similar capillary structures formation, however ECFC from inactive GPA alone showed early loss of angiogenic capacity between 15 and 24 hours. In scratching assay, there was an impairment in the proliferative capacity of the ECFC between GPA patients and controls without significant difference (12th hour, p = 0.05). When incubated with control plasma, ECFC of remission GPA patients showed a significant lower migration capacity after the 4th hour (p = 0.0001). In longitudinal analysis, ECFC isolated after treatment showed significantly lower migration rates.
CONCLUSIONS
PR3-positive remission GPA ECFC demonstrated early loss of tube formation and less proliferative capacity compared to those of healthy controls, suggesting impairment of endothelial function.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5