Background
Diseases associated with the circulatory system are the main causes of worldwide morbidity and mortality, implying the need for vascular implants. Thus, the production of vascular biomaterials has proven to be a promising alternative to therapies used in studies and research related to vascular physiology.
Objectives
The present project aims to achieve the artificial development of blood vessels through the recellularization of vascular scaffolds derived from bovine placental vessels.
Methods
The chorioallantoic surface of the bovine placenta was used to produce decellularized biomaterials. For recellularization, 2.5 x 104 endothelial cells were seeded above each decellularized vessel fragment during three or seven days, when culture were interrupted, and the fragments were fixed for cell attachment analysis. Decellularized and recellularized biomaterials were evaluated by basic histology, scanning electron microscopy, and immunohistochemistry.
Results
The decellularization process produced vessels that maintained natural structure and elastin content, and no cells or gDNA remains were observed. Endothelial precursor cells were also attached to lumen and external surface of the decellularized vessel.Conclusion: Our results show a possibility of future uses of this biomaterial in cardiovascular medicine, as in the development of engineered vessels.
Decellularized Extracellular Matrix; Bioengineering; Blood Vessels
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Analysis of decellularized bovine placental vessel. Hematoxylin eosin, (A1) Native vessel, Bar= 20 μm, (^) lumen: extracellular matrix fibers stained in pink and cell nucleus in purple; (B1) Decellularized, Bar= 20 μm,(*) external surface: absent cell nuclei; (C-F1) Recellularized, Bar= 20 μm, (*) external surface: (C1), Recellularized three days (SV line), cell nuclei stained in purple adhered to the external surface of the biomaterial; (D1), Recellularized three days with canine yolk sac cells with vascular endothelial growth factor (VEGF) and enhanced green fluorescent protein (eGFP) overexpression (SV-VEGF lineage), extracellular matrix fibers stained in pink and cell nuclei in purple adhered to the external surface of the biomaterial; (E1), Recellularized seven days canine yolk sac cells (SV-lineage), cell nuclei stained in purple adhered to the external surface of the biomaterial; (F1), Recellularized seven days with cells (SV-VEGF lineage), extracellular matrix fibers stained in pink, cell nuclei in purple adhered to the external surface of the biomaterial. (A4) bar = 2 μm, decellularized: Observe preservation of fibers of the extracellular matrix of the vascular biomaterial; Recellularization with three days of culture (B4, C4): (B4) bar = 30 μm, with SV cells adhered to the vascular biomaterial wall; (C4) bar = 10 μm, SV-VEGF cells adhered to the vascular biomaterial wall. Immunohistochemistry assay: elastin expression in the vascular biomaterial of the native bovine chorioallantoic surface, and recellularized with SV-VEGF cells. Caption: Bar=20 µm, (A3) native blood vessel of the chorioallantoic surface of the bovine placenta, observe elastin expression in the entire vascular wall; (B3) native blood vessel from the chorioallantoic surface of the bovine placenta, negative control stained with hematoxylin; (C3) 7-day recellularization of bovine placental vascular biomaterial with SV-VEGF cells, observing elastin expression in the entire vascular wall; (D3) 7-day recellularization of bovine placental vascular biomaterial with SV-VEGF cells, negative control stained with hematoxylin. Recellularizarion assay: Bar 20 µm ; (A2-C2) 3 days of cultivation; (A2) decellularized control vascular biomaterial, observe absent nuclei; (B2) biomaterial recellularized with canine yolk sac endothelial progenitor cells (SV), observe nuclei stained in blue by 4’,6’-diamino-2-fenil-indol (DAPI); (C2) recellularized biomaterial with SV-VEGF cells, observe nuclei stained blue by DAPI, eGFP not observed; (D2-F2) 7 days of cultivation; (D2) vascular biomaterial decellularized control, observe absent nuclei; (E2) biomaterial recellularized with SV cells, observe nuclei stained in blue by DAPI; (F2) Biomaterial recellularized with SV-VEGF cells, observe nuclei stained in blue by DAPI.
