title: Functions of corneal endothelial cells do not change after uptake of superparamagnetic iron oxide nanoparticles.
abstract: To avoid donor tissue shortages, ex vivo cultured human corneal endothelial cell (HCEC) transplantation is a promising therapeutic resource. Superparamagnetic iron oxide nanoparticle (SPION) cell labeling assists HCEC transplantation by attaching the posterior corneal stroma in ex vivo animal models. However, possible functional changes of the HCECs following SPION labeling remain to be determined. In this study, we used SPIONs to label cultured rabbit CECs (RCECs) in order to observe important cell functions and the levels of cell markers. The synthetic SPIONs exhibited superparamagnetism at room temperature, with saturation magnetization of 55.4 emu/g and negligible remanence or coercivity. The ζ-potential was -24.5 mV and the diameter was 101 ± 55 nm. Immunostaining demonstrated a normal density of zonula occluden-1 (ZO-1), nestin and Ki-67 at cellular junctions or in nuclei from RCECs following SPION labeling at 16 µg/ml. MTT cytotoxicity assay, homotypic adhesion assay, quantitative flow cytometric Ki-67 analysis and RCEC pump function measurement demonstrated no significant differences between the cells with or without SPION labeling (P<0.05, for all assays). Results of this study demonstrated successful labeled cultured RCECs with synthetic SPIONs. Labeled cells possessed several important characteristics required to maintain the transparency and refractive parameters of the cornea, including hexagonal cell morphology, higher cell adhesion ability and proliferative potential, cell pump function and the positive expression of several cell markers.
authors.affiliation: Department of Ophthalmology, Tongji Hospital Affiliated to Tongji University School of Medicine, Shanghai 200065, P.R. China.
authors.forename: Yan-Long
authors.initials: YL
authors.lastname: Bi
authors.forename: Ming-Feng
authors.initials: MF
authors.lastname: Wu
authors.forename: Li-Xia
authors.initials: LX
authors.lastname: Lu
authors.forename: Qi
authors.initials: Q
authors.lastname: Zhou
authors.forename: Fei
authors.initials: F
authors.lastname: Du
authors.forename: Xiao-Ting
authors.initials: XT
authors.lastname: Sun
authors.forename: Sen-Fei
authors.initials: SF
authors.lastname: Tang
authors.forename: Guo-Tong
authors.initials: GT
authors.lastname: Xu
mesh_terms: D000818:Animals; D002448:Cell Adhesion / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003911:Dextrans / Q000737:chemistry; D004195:Disease Models, Animal; D004728:Endothelium, Corneal / Q000166:cytology / Q000378:metabolism*; D005290:Ferric Compounds / Q000737:chemistry*; D007381:Intermediate Filament Proteins / Q000378:metabolism; D019394:Ki-67 Antigen / Q000378:metabolism; D008280:Magnetics; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000633:toxicity; D009419:Nerve Tissue Proteins / Q000378:metabolism; D064231:Nestin; D011817:Rabbits; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D062826:Zonula Occludens-1 Protein / Q000378:metabolism
chemical_list: D003911:Dextrans; D005290:Ferric Compounds; D007381:Intermediate Filament Proteins; D019394:Ki-67 Antigen; C577484:NES protein, human; D009419:Nerve Tissue Proteins; D064231:Nestin; D062826:Zonula Occludens-1 Protein; C000499:ferric oxide; D000254:Sodium-Potassium-Exchanging ATPase