Perfusion Decellularization of Extrahepatic Bile Duct Allows Tissue-Engineered Scaffold Generation by Preserving Matrix Architecture and Cytocompatibility


Por: Ramirez-Marin, Yolik, Abad-Contreras, David Eduardo, Ustarroz-Cano, Martha, Perez-Gallardo, Norma S., Villafuerte-Garcia, Lorena, Puente-Guzman, Dulce Maria, Villar-Velasco, Jorge Luna del, Rodriguez-Lopez, Leonardo Alejandro, Torres-Villalobos, Gonzalo, MERCADO, MIGUEL ANGEL, Tapia-Jurado, Jesus, Martinez-Garcia, Francisco Drusso, Harmsen, Martin Conrad, Pina-Barba, M. Cristina, Giraldo-Gomez, David M.

Publicada: 1 jun 2021
Resumen:
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.

Filiaciones:
Ramirez-Marin, Yolik:
 Natl Autonomous Univ Mexico UNAM, Fac Med, Div Posgrad Studies, Program Med Specializat Gen Surg, Ave Univ 3000,Unidad Posgrad Edificio E 2 Piso, Mexico City 04510, DF, Mexico

 Natl Inst Med Sci & Nutr Mexico Salvador Zubiran, Vasco Quiroga 15,Belisario Dominguez Secc 16, Mexico City 14080, DF, Mexico

Abad-Contreras, David Eduardo:
 Natl Autonomous Univ Mexico UNAM, Mat Res Inst, Lab Biomat, Ave Univ 3000, Mexico City 04510, DF, Mexico

Ustarroz-Cano, Martha:
 Natl Autonomous Univ Mexico UNAM, Dept Cell & Tissue Biol, Fac Med, Ave Univ 3000,Edificio 3 Piso,Ciudad Univ, Mexico City 04510, DF, Mexico

Perez-Gallardo, Norma S.:
 Natl Autonomous Univ Mexico UNAM, Fac Vet Med & Anim Husb, Surg Training Sect, Ave Univ 3000,Ciudad Univ, Mexico City 04510, DF, Mexico

Villafuerte-Garcia, Lorena:
 Natl Autonomous Univ Mexico UNAM, Fac Vet Med & Anim Husb, Surg Training Sect, Ave Univ 3000,Ciudad Univ, Mexico City 04510, DF, Mexico

Puente-Guzman, Dulce Maria:
 Natl Autonomous Univ Mexico UNAM, Fac Vet Med & Anim Husb, Surg Training Sect, Ave Univ 3000,Ciudad Univ, Mexico City 04510, DF, Mexico

Villar-Velasco, Jorge Luna del:
 Natl Autonomous Univ Mexico UNAM, Fac Vet Med & Anim Husb, Surg Training Sect, Ave Univ 3000,Ciudad Univ, Mexico City 04510, DF, Mexico

Rodriguez-Lopez, Leonardo Alejandro:
 Natl Inst Med Sci & Nutr Mexico Salvador Zubiran, Vasco Quiroga 15,Belisario Dominguez Secc 16, Mexico City 14080, DF, Mexico

Torres-Villalobos, Gonzalo:
 Natl Inst Med Sci & Nutr Mexico Salvador Zubiran, Vasco Quiroga 15,Belisario Dominguez Secc 16, Mexico City 14080, DF, Mexico

MERCADO, MIGUEL ANGEL:
 Natl Inst Med Sci & Nutr Mexico Salvador Zubiran, Vasco Quiroga 15,Belisario Dominguez Secc 16, Mexico City 14080, DF, Mexico

Tapia-Jurado, Jesus:
 Natl Autonomous Univ Mexico UNAM, Fac Med, Div Posgrad Studies, Unit Adv Med Simulat, Ave Univ 3000,Unidad Posgrad Edificio B 2 Piso, Mexico City 04510, DF, Mexico

Martinez-Garcia, Francisco Drusso:
 Univ Groningen, Univ Med Ctr Groningen, Dept Pathol & Med Biol, Hanzepl 1, NL-9713 Groningen, Netherlands

Harmsen, Martin Conrad:
 Univ Groningen, Univ Med Ctr Groningen, Dept Pathol & Med Biol, Hanzepl 1, NL-9713 Groningen, Netherlands

Pina-Barba, M. Cristina:
 Natl Autonomous Univ Mexico UNAM, Mat Res Inst, Lab Biomat, Ave Univ 3000, Mexico City 04510, DF, Mexico

Giraldo-Gomez, David M.:
 Natl Autonomous Univ Mexico UNAM, Dept Cell & Tissue Biol, Fac Med, Ave Univ 3000,Edificio 3 Piso,Ciudad Univ, Mexico City 04510, DF, Mexico

 Natl Autonomous Univ Mexico UNAM, Fac Med, Microscopy Core Facil, Ave Univ 3000,Edificio A Planta Baja, Mexico City 04510, DF, Mexico
ISSN: 19961944
Editorial
MDPI AG, POSTFACH, CH-4005 BASEL, SWITZERLAND, Suiza
Tipo de documento: Article
Volumen: 14 Número: 11
Páginas:
WOS Id: 000660269000001
ID de PubMed: 34198787
imagen Gold, Green Published, All Open Access, Green

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