title: Polydimethylsiloxane as a substrate for retinal pigment epithelial cell growth.
abstract: Retinal pigment epithelial (RPE) cell transplantation represents potential treatment for age-related macular degeneration (AMD). Because delivery of isolated cells can cause serious complications, it is necessary to develop a suitable transplant membrane that could support an intact functioning RPE monolayer. Polydimethylsiloxane (PDMS) possesses the physical properties required for a transplanting device and is widely used clinically. We have investigated the use of PDMS as a potential surface for the growth of healthy RPE monolayers. PDMS discs were surface modified by air and ammonia gas plasma treatments. Dynamic contact angles were measured to determine the changes in wettability. Human ARPE-19 cells were seeded onto untreated and treated samples. Cell number, morphology and monolayer formation, cytotoxicity, and phagocytosis of photoreceptor outer segments (POS) were assessed at set time-points. Air plasma treatment increased the wettability of PDMS. This significantly enhanced cell growth, reaching confluence by day 7. Immunofluorescence revealed well-defined actin staining, monolayer formation, and high cell viability on air plasma treated and untreated surfaces, and to a lesser extent, on ammonia plasma treated. Furthermore, RPE monolayers were able to demonstrate phagocytosis of POS in a time-dependent manner similar to control. PDMS can support an intact functional monolayer of healthy differentiated RPE cells.
authors.affiliation: Department of Ophthalmology, School of Clinical Sciences, The University of Liverpool, Liverpool L69 3GA, United Kingdom. y.krishna@liverpool.ac.uk
authors.forename: Yamini
authors.initials: Y
authors.lastname: Krishna
authors.forename: Carl M
authors.initials: CM
authors.lastname: Sheridan
authors.forename: David L
authors.initials: DL
authors.lastname: Kent
authors.forename: Ian
authors.initials: I
authors.lastname: Grierson
authors.forename: Rachel L
authors.initials: RL
authors.lastname: Williams
mesh_terms: D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D004129:Dimethylpolysiloxanes / Q000494:pharmacology* / Q000627:therapeutic use; D004847:Epithelial Cells; D006801:Humans; D007700:Kinetics; D010587:Phagocytosis / Q000187:drug effects; D010857:Pigment Epithelium of Eye / Q000166:cytology* / Q000187:drug effects; D012374:Rod Cell Outer Segment / Q000276:immunology; D012828:Silicones / Q000494:pharmacology* / Q000627:therapeutic use; D013499:Surface Properties; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D004129:Dimethylpolysiloxanes; D012828:Silicones; C013830:baysilon