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Characterization of Endothelial Progenitor Cell Interactions with Human Tropoelastin.

Yu Y, Wise SG, Michael PL, Bax DV, Yuen GS, Hiob MA, Yeo GC, Filipe EC, Dunn LL, Chan KH, Hajian H, Celermajer DS, Weiss AS, Ng MK - PLoS ONE (2015)

Bottom Line: The rapid restoration of a functional endothelium is known to reduce these complications.Integrin αVβ3 blocking antibodies decreased EPC adhesion and spreading on rhTE by 39±3% and 56±10% respectively, demonstrating a large contribution from this specific integrin.In conclusion, rhTE supports EPC binding via an integrin mechanism involving αVβ3.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Royal Prince Alfred Hospital, Sydney, NSW, 2050, Australia; The Heart Research Institute, Sydney, NSW, 2042, Australia; Sydney Medical School, University of Sydney, Sydney, NSW, 2006, Australia.

ABSTRACT
The deployment of endovascular implants such as stents in the treatment of cardiovascular disease damages the vascular endothelium, increasing the risk of thrombosis and promoting neointimal hyperplasia. The rapid restoration of a functional endothelium is known to reduce these complications. Circulating endothelial progenitor cells (EPCs) are increasingly recognized as important contributors to device re-endothelialization. Extracellular matrix proteins prominent in the vessel wall may enhance EPC-directed re-endothelialization. We examined attachment, spreading and proliferation on recombinant human tropoelastin (rhTE) and investigated the mechanism and site of interaction. EPCs attached and spread on rhTE in a dose dependent manner, reaching a maximal level of 56±3% and 54±3%, respectively. EPC proliferation on rhTE was comparable to vitronectin, fibronectin and collagen. EDTA, but not heparan sulfate or lactose, reduced EPC attachment by 81±3%, while full attachment was recovered after add-back of manganese, inferring a classical integrin-mediated interaction. Integrin αVβ3 blocking antibodies decreased EPC adhesion and spreading on rhTE by 39±3% and 56±10% respectively, demonstrating a large contribution from this specific integrin. Attachment of EPCs on N-terminal rhTE constructs N25 and N18 accounted for most of this interaction, accompanied by comparable spreading. In contrast, attachment and spreading on N10 was negligible. αVβ3 blocking antibodies reduced EPC spreading on both N25 and N18 by 45±4% and 42±14%, respectively. In conclusion, rhTE supports EPC binding via an integrin mechanism involving αVβ3. N25 and N18, but not N10 constructs of rhTE contribute to EPC binding. The regulation of EPC activity by rhTE may have implications for modulation of the vascular biocompatibility of endovascular implants.

No MeSH data available.


Related in: MedlinePlus

Schematic diagram showing full-length rhTE and constructs N18, N25 and N10.Key domain features are indicated.
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pone.0131101.g001: Schematic diagram showing full-length rhTE and constructs N18, N25 and N10.Key domain features are indicated.

Mentions: Full-length recombinant human tropoelastin (rhTE) and N-terminal constructs representing the first 25 (N25), 18 (N18), and 10 (N10) domains (Fig 1) of rhTE were produced in a previously described E. coli expression system [34]. Human dermal fibroblasts (HDF; line GM3348) were obtained from Coriell Research Institute (Camden, NJ, USA). Primary human coronary artery smooth muscle cells (SMCs) were obtained from Cell Applications (San Diego, CA, USA). Fibronectin and vitronectin from human plasma, as well as calf skin collagen type 1 (Sigma) were used as controls. Anti-human integrin antibodies, αvβ3-clone LM609, α5β1-clone JBS5, and α2β1-clone BHA2.1 were obtained from Millipore. All other reagents were purchased from Sigma.


Characterization of Endothelial Progenitor Cell Interactions with Human Tropoelastin.

Yu Y, Wise SG, Michael PL, Bax DV, Yuen GS, Hiob MA, Yeo GC, Filipe EC, Dunn LL, Chan KH, Hajian H, Celermajer DS, Weiss AS, Ng MK - PLoS ONE (2015)

Schematic diagram showing full-length rhTE and constructs N18, N25 and N10.Key domain features are indicated.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4482626&req=5

pone.0131101.g001: Schematic diagram showing full-length rhTE and constructs N18, N25 and N10.Key domain features are indicated.
Mentions: Full-length recombinant human tropoelastin (rhTE) and N-terminal constructs representing the first 25 (N25), 18 (N18), and 10 (N10) domains (Fig 1) of rhTE were produced in a previously described E. coli expression system [34]. Human dermal fibroblasts (HDF; line GM3348) were obtained from Coriell Research Institute (Camden, NJ, USA). Primary human coronary artery smooth muscle cells (SMCs) were obtained from Cell Applications (San Diego, CA, USA). Fibronectin and vitronectin from human plasma, as well as calf skin collagen type 1 (Sigma) were used as controls. Anti-human integrin antibodies, αvβ3-clone LM609, α5β1-clone JBS5, and α2β1-clone BHA2.1 were obtained from Millipore. All other reagents were purchased from Sigma.

Bottom Line: The rapid restoration of a functional endothelium is known to reduce these complications.Integrin αVβ3 blocking antibodies decreased EPC adhesion and spreading on rhTE by 39±3% and 56±10% respectively, demonstrating a large contribution from this specific integrin.In conclusion, rhTE supports EPC binding via an integrin mechanism involving αVβ3.

View Article: PubMed Central - PubMed

Affiliation: Department of Cardiology, Royal Prince Alfred Hospital, Sydney, NSW, 2050, Australia; The Heart Research Institute, Sydney, NSW, 2042, Australia; Sydney Medical School, University of Sydney, Sydney, NSW, 2006, Australia.

ABSTRACT
The deployment of endovascular implants such as stents in the treatment of cardiovascular disease damages the vascular endothelium, increasing the risk of thrombosis and promoting neointimal hyperplasia. The rapid restoration of a functional endothelium is known to reduce these complications. Circulating endothelial progenitor cells (EPCs) are increasingly recognized as important contributors to device re-endothelialization. Extracellular matrix proteins prominent in the vessel wall may enhance EPC-directed re-endothelialization. We examined attachment, spreading and proliferation on recombinant human tropoelastin (rhTE) and investigated the mechanism and site of interaction. EPCs attached and spread on rhTE in a dose dependent manner, reaching a maximal level of 56±3% and 54±3%, respectively. EPC proliferation on rhTE was comparable to vitronectin, fibronectin and collagen. EDTA, but not heparan sulfate or lactose, reduced EPC attachment by 81±3%, while full attachment was recovered after add-back of manganese, inferring a classical integrin-mediated interaction. Integrin αVβ3 blocking antibodies decreased EPC adhesion and spreading on rhTE by 39±3% and 56±10% respectively, demonstrating a large contribution from this specific integrin. Attachment of EPCs on N-terminal rhTE constructs N25 and N18 accounted for most of this interaction, accompanied by comparable spreading. In contrast, attachment and spreading on N10 was negligible. αVβ3 blocking antibodies reduced EPC spreading on both N25 and N18 by 45±4% and 42±14%, respectively. In conclusion, rhTE supports EPC binding via an integrin mechanism involving αVβ3. N25 and N18, but not N10 constructs of rhTE contribute to EPC binding. The regulation of EPC activity by rhTE may have implications for modulation of the vascular biocompatibility of endovascular implants.

No MeSH data available.


Related in: MedlinePlus