Perfusion Decellularization of Extrahepatic Bile Duct Allows Tissue-Engineered Scaffold Generation by Preserving Matrix Architecture and Cytocompatibility
Yolik Ramírez-Marín,
David Eduardo Abad-Contreras,
Martha Ustarroz-Cano,
Norma S. Pérez-Gallardo,
Lorena Villafuerte-García,
Dulce Maria Puente-Guzmán,
Jorge Luna del Villar-Velasco,
Leonardo Alejandro Rodríguez-López,
Gonzalo Torres-Villalobos,
Miguel Ángel Mercado,
Jesús Tapia-Jurado,
Francisco Drusso Martínez-García,
Martin Conrad Harmsen,
M. Cristina Piña-Barba,
David M. Giraldo-Gomez
Affiliations
Yolik Ramírez-Marín
Program of Medical Specialization General Surgery, Division of Posgraduate Studies, Faculty of Medicine, National Autonomous University of Mexico (UNAM), Avenida Universidad 3000, Circuito de Posgrados, Unidad de Posgrado Edificio “E” 2° piso, Ciudad Universitaria, Coyoacán, Ciudad de México 04510, Mexico
David Eduardo Abad-Contreras
Laboratory for Biomaterials, Materials Research Institute, National Autonomous University of Mexico (UNAM), Avenida Universidad 3000, Circuito Exterior, Ciudad Universitaria, Coyoacán, Ciudad de México 04510, Mexico
Martha Ustarroz-Cano
Department of Cell and Tissue Biology, Faculty of Medicine, National Autonomous University of Mexico (UNAM), Avenida Universidad 3000, Circuito Interior, Edificio “A” 3° piso, Ciudad Universitaria, Coyoacán, Ciudad de México 04510, Mexico
Norma S. Pérez-Gallardo
Surgical Training Section, Faculty of Veterinary Medicine and Animal Husbandry, National Autonomous University of Mexico (UNAM), Avenida Universidad 3000, Circuito Exterior, Ciudad Universitaria, Coyoacán, Ciudad de México 04510, Mexico
Lorena Villafuerte-García
Surgical Training Section, Faculty of Veterinary Medicine and Animal Husbandry, National Autonomous University of Mexico (UNAM), Avenida Universidad 3000, Circuito Exterior, Ciudad Universitaria, Coyoacán, Ciudad de México 04510, Mexico
Dulce Maria Puente-Guzmán
Surgical Training Section, Faculty of Veterinary Medicine and Animal Husbandry, National Autonomous University of Mexico (UNAM), Avenida Universidad 3000, Circuito Exterior, Ciudad Universitaria, Coyoacán, Ciudad de México 04510, Mexico
Jorge Luna del Villar-Velasco
Surgical Training Section, Faculty of Veterinary Medicine and Animal Husbandry, National Autonomous University of Mexico (UNAM), Avenida Universidad 3000, Circuito Exterior, Ciudad Universitaria, Coyoacán, Ciudad de México 04510, Mexico
Leonardo Alejandro Rodríguez-López
National Institute of Medical Sciences and Nutrition of Mexico Salvador Zubirán, Vasco de Quiroga 15, Belisario Domínguez Secc. 16, Tlalpan, Ciudad de México 14080, Mexico
Gonzalo Torres-Villalobos
National Institute of Medical Sciences and Nutrition of Mexico Salvador Zubirán, Vasco de Quiroga 15, Belisario Domínguez Secc. 16, Tlalpan, Ciudad de México 14080, Mexico
Miguel Ángel Mercado
National Institute of Medical Sciences and Nutrition of Mexico Salvador Zubirán, Vasco de Quiroga 15, Belisario Domínguez Secc. 16, Tlalpan, Ciudad de México 14080, Mexico
Jesús Tapia-Jurado
Unit of Advanced Medical Simulation, Division of Posgraduate Studies, Faculty of Medicine, National Autonomous University of Mexico (UNAM), Avenida Universidad 3000, Circuito de Posgrados, Unidad de Posgrado Edificio “B” 2° piso, Ciudad Universitaria, Coyoacán, Ciudad de México 04510, Mexico
Francisco Drusso Martínez-García
Department of Pathology and Medical Biology, University Medical Center Groningen University of Groningen, Hanzeplein 1, 9713 Groningen, The Netherlands
Martin Conrad Harmsen
Department of Pathology and Medical Biology, University Medical Center Groningen University of Groningen, Hanzeplein 1, 9713 Groningen, The Netherlands
M. Cristina Piña-Barba
Laboratory for Biomaterials, Materials Research Institute, National Autonomous University of Mexico (UNAM), Avenida Universidad 3000, Circuito Exterior, Ciudad Universitaria, Coyoacán, Ciudad de México 04510, Mexico
David M. Giraldo-Gomez
Department of Cell and Tissue Biology, Faculty of Medicine, National Autonomous University of Mexico (UNAM), Avenida Universidad 3000, Circuito Interior, Edificio “A” 3° piso, Ciudad Universitaria, Coyoacán, Ciudad de México 04510, Mexico
Reconstruction of bile ducts damaged remains a vexing medical problem. Surgeons have few options when it comes to a long segment reconstruction of the bile duct. Biological scaffolds of decellularized biliary origin may offer an approach to support the replace of bile ducts. Our objective was to obtain an extracellular matrix scaffold derived from porcine extrahepatic bile ducts (dECM-BD) and to analyze its biological and biochemical properties. The efficiency of the tailored perfusion decellularization process was assessed through histology stainings. Results from 4’-6-diamidino-2-phenylindole (DAPI), Hematoxylin and Eosin (H&E) stainings, and deoxyribonucleic acid (DNA) quantification showed proper extracellular matrix (ECM) decellularization with an effectiveness of 98%. Immunohistochemistry results indicate an effective decrease in immunogenic marker as human leukocyte antigens (HLA-A) and Cytokeratin 7 (CK7) proteins. The ECM of the bile duct was preserved according to Masson and Herovici stainings. Data derived from scanning electron microscopy (SEM) and thermogravimetric analysis (TGA) showed the preservation of the dECM-BD hierarchical structures. Cytotoxicity of dECM-BD was null, with cells able to infiltrate the scaffold. In this work, we standardized a decellularization method that allows one to obtain a natural bile duct scaffold with hierarchical ultrastructure preservation and adequate cytocompatibility.
