title: The basement membrane of the isolated rat colonic mucosa. A light, electron and atomic force microscopy study.
abstract: Basement membranes (BM) are structures of the extracellular matrix (ECM), which are involved in epithelial barriers, but also play an important role in processes such as cell adhesion, cell growth and tissue healing. The aim of this study was to investigate possible effects of cell removal on the structure of the BM of the colonic mucosa. The superficial epithelium was removed with EDTA and the samples were then mechanically fixed for immunohistochemistry, TEM, SEM and AFM. For SEM and AFM, some samples were also prepared according to the OTO method. BM marker proteins were detected after cell removal by immunohistochemistry, indicating that BM remains. However, a lamina lucida (LL) was no longer visible in TEM, it disappeared and the BM became slightly thinner. The surface topography of the BM is characterized by the presence of globules, fenestrations and pore-like structures, which were visualized with SEM and AFM. Noteworthy is the visualization for the first time with AFM of a 3D network of fine fibers and filaments ("cords"), which very much resembled that described with TEM by Inoue (1994). An unresolved question is whether the pore-like structures observed in this study, especially with SEM, actually correspond to the pores of the BM whose existence has been demonstrated functionally. In conclusion, the structural patterns and changes described could be considered as a reference to evaluate the effects of other decellularization protocols on BMs, such as those used in tissue engineering.
authors.affiliation: Department of Histology and Pathology, Faculty for Health Sciences, University Rey Juan Carlos I, 28922 Alcorcon, Madrid, Spain; Department of Anatomy and Cell Biology, Medical School, Saarland University, 66421 Homburg Saar, Germany. Electronic address: pedro.mestres@urjc.es.
authors.forename: Pedro
authors.initials: P
authors.lastname: Mestres
authors.affiliation: Department of Histology and Pathology, Faculty for Health Sciences, University Rey Juan Carlos I, 28922 Alcorcon, Madrid, Spain.
authors.forename: Laura Lopez
authors.initials: LL
authors.lastname: Gomez
authors.affiliation: Department of Histology and Pathology, Faculty for Health Sciences, University Rey Juan Carlos I, 28922 Alcorcon, Madrid, Spain.
authors.forename: Teresa Nuñez
authors.initials: TN
authors.lastname: Lopez
authors.affiliation: Laboratory for Electron Microscopy, Centre for Technical Support (CAT), University Rey Juan Carlos I, 28933 Mostoles, Madrid, Spain.
authors.forename: Gilberto
authors.initials: G
authors.lastname: del Rosario
authors.affiliation: Department of Experimental Physics, Saarland University, 66041 Saarbrucken, Germany.
authors.forename: Slavomir Witold
authors.initials: SW
authors.lastname: Lukas
authors.affiliation: Department of Experimental Physics, Saarland University, 66041 Saarbrucken, Germany.
authors.forename: Uwe
authors.initials: U
authors.lastname: Hartmann
mesh_terms: D000818:Animals; D001485:Basement Membrane / Q000648:ultrastructure*; D003106:Colon / Q000648:ultrastructure*; D020895:Enterocytes / Q000648:ultrastructure; D005724:Ganglia / Q000033:anatomy & histology; D007413:Intestinal Mucosa / Q000294:innervation / Q000648:ultrastructure*; D008565:Membrane Proteins / Q000737:chemistry; D018625:Microscopy, Atomic Force; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D051381:Rats; D017208:Rats, Wistar
chemical_list: D008565:Membrane Proteins
keywords: AFM; Basement membrane; Colon; Decellularization; Scanning electron microscopy; Tissue preparation methods; Transmission electron microscopy