title: Tissue-engineered blood vessel substitute by reconstruction of endothelium using mesenchymal stem cells induced by platelet growth factors.
abstract: BACKGROUND
  Cardiovascular diseases remain leaders as the major causes of mortality in Western society. Restoration of the circulation through construction of bypass surgical treatment is regarded as the gold standard treatment of peripheral vascular diseases, and grafts are necessary for this purpose. The great saphenous vein is often not available and synthetic grafts have their limitations. Therefore, new techniques to produce alternative grafts have been developed and, in this sense, tissue engineering is a promising alternative to provide biocompatible grafts. This study objective was to reconstruct the endothelium layer of decellularized vein scaffolds, using mesenchymal stem cells (MSCs) and growth factors obtained from platelets.
  
  
  METHODS
  Fifteen nonpregnant female adult rabbits were used for all experiments. Adipose tissue and vena cava were obtained and subjected to MSCs isolation and tissue decellularization, respectively. MSCs were subjected to differentiation using endothelial inductor growth factor (EIGF) obtained from human platelet lysates. Immunofluorescence, histological and immunohistochemical analyses were employed for the final characterization of the obtained blood vessel substitute.
  
  
  RESULTS
  The scaffolds were successfully decellularized with sodium dodecyl sulfate. MSCs actively adhered at the scaffolds, and through stimulation with EIGF were differentiated into functional endothelial cells, secreting significantly higher quantities of von Willebrand factor (0.85 μg/mL; P < .05) than cells cultivated under the same conditions, without EIGF (0.085 μg/mL). Cells with evident morphologic characteristics of endothelium were seen at the lumen of the scaffolds. These cells also stained positive for fascin protein, which is highly expressed by differentiated endothelial cells.
  
  
  CONCLUSIONS
  Taken together, the use of decellularized bioscaffold and subcutaneous MSCs seems to be a potential approach to obtain bioengineered blood vessels, in the presence of EIGF supplementation.
authors.affiliation: Department of Surgery and Orthopedics, Vascular Laboratory, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil; Cell Engineering Laboratory, Blood Transfusion Center, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil. Electronic address: matheus.fameca@ig.com.br.
authors.forename: Matheus
authors.initials: M
authors.lastname: Bertanha
authors.affiliation: Cell Engineering Laboratory, Blood Transfusion Center, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil; Department of Morphology, Extracellular Matrix Laboratory, Botucatu Biosciences Institute, UNESP-Paulista State University, Botucatu, Brazil.
authors.forename: Andrei
authors.initials: A
authors.lastname: Moroz
authors.affiliation: Cell Engineering Laboratory, Blood Transfusion Center, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Almeida
authors.affiliation: Cell Engineering Laboratory, Blood Transfusion Center, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil.
authors.forename: Flavia Cilene
authors.initials: FC
authors.lastname: Alves
authors.affiliation: Cell Engineering Laboratory, Blood Transfusion Center, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil.
authors.forename: Michele Janegitz
authors.initials: MJ
authors.lastname: Acorci Valério
authors.affiliation: Department of Surgery and Orthopedics, Vascular Laboratory, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil.
authors.forename: Regina
authors.initials: R
authors.lastname: Moura
authors.affiliation: Department of Pathology, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil.
authors.forename: Maria Aparecida Custódio
authors.initials: MA
authors.lastname: Domingues
authors.affiliation: Department of Surgery and Orthopedics, Vascular Laboratory, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil.
authors.forename: Marcone Lima
authors.initials: ML
authors.lastname: Sobreira
authors.affiliation: Cell Engineering Laboratory, Blood Transfusion Center, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil; Department of Urology, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil.
authors.forename: Elenice
authors.initials: E
authors.lastname: Deffune
mesh_terms: D000818:Animals; D001807:Blood Vessel Prosthesis; D002454:Cell Differentiation; D002478:Cells, Cultured; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000166:cytology*; D005260:Female; D006801:Humans; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D010982:Platelet-Derived Growth Factor / Q000378:metabolism*; D011817:Rabbits; D019651:Plastic Surgery Procedures / Q000379:methods*; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds
chemical_list: D010982:Platelet-Derived Growth Factor