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
Gold, Green Published, All Open Access, Green
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