Matheus Bertanha1, Andrei Moroz2, Rodrigo Almeida3, Flavia Cilene Alves3, Michele Janegitz Acorci Valério3, Regina Moura4, Maria Aparecida Custódio Domingues5, Marcone Lima Sobreira4, Elenice Deffune6. 1. 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. 2. 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. 3. Cell Engineering Laboratory, Blood Transfusion Center, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil. 4. Department of Surgery and Orthopedics, Vascular Laboratory, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil. 5. Department of Pathology, Botucatu Medical School, UNESP-Paulista State University, Botucatu, Brazil. 6. 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.
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.
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: Diego Noe Rodriguez Sanchez; Matheus Bertanha; Thiago Dias Fernandes; Luiz Antônio de Lima Resende; Elenice Deffune; Rogério Martins Amorim Journal: Int J Stem Cells Date: 2017-05-30 Impact factor: 2.500