Development of an esophagus acellular matrix tissue scaffold.
A cell-extraction protocol yielding an esophagus acellular matrix (EAM) scaffold for use in tissue engineering of an esophagus, including hypotonic lysis, multiple detergent cell extraction steps, and nucleic acid digestion, was developed in a rat model. Histological techniques, burst pressure studies, in vitro esophageal epithelial cell seeding, and in vivo implantation were used to assess cell extraction, extracellular matrix (ECM) preservation, and biocompatibility. Microscopy demonstrated that cell extraction protocols using sodium dodecyl sulfate (SDS) (0.5%, wt/vol) as a detergent resulted in cell-free EAM with retained ECM protein collagen, elastin, laminin, and fibronectin. Burst pressure studies indicated a loss of tensile strength in EAMs, but at intraluminal pressures that were unlikely to affect in vivo application. In vitro cell seeding studies exhibited epithelial cell proliferation with stratification similar to native esophagi after 11 days, and subcutaneously implanted EAMs displayed neovascularization and a minimal inflammatory response after 30 days of implantation. This study presents an esophagus acellular matrix tissue scaffold with preserved ECM proteins, biomechanical properties, and the ability to support esophageal cell proliferation to serve as the foundation for a tissue-engineered esophagus.
Department of Otolaryngology-Head and Neck Surgery, University of Washington, Seattle, USA.
Amit D
AD
Bhrany
Benjamin L
BL
Beckstead
Tess C
TC
Lang
D Gregory
DG
Farwell
Cecilia M
CM
Giachelli
Buddy D
BD
Ratner
D000818:Animals; D001672:Biocompatible Materials; D001696:Biomechanical Phenomena; D003094:Collagen / Q000378:metabolism / Q000648:ultrastructure; D004848:Epithelium / Q000187:drug effects / Q000648:ultrastructure; D004947:Esophagus / Q000737:chemistry* / Q000648:ultrastructure; D005109:Extracellular Matrix / Q000378:metabolism* / Q000648:ultrastructure; D005353:Fibronectins / Q000378:metabolism / Q000648:ultrastructure; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes; D006651:Histocytochemistry; D020007:Implants, Experimental; D007211:Indoles; D007797:Laminin / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D011916:Rats, Inbred F344; D011917:Rats, Inbred Lew; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013501:Surface-Active Agents / Q000494:pharmacology; D023822:Tissue Engineering / Q000379:methods; D016042:Transplantation, Heterotopic
D001672:Biocompatible Materials; D005353:Fibronectins; D005456:Fluorescent Dyes; D007211:Indoles; D007797:Laminin; D013501:Surface-Active Agents; D012967:Sodium Dodecyl Sulfate; C007293:DAPI; D003094:Collagen