title: Attachment of human primary osteoblast cells to modified polyethylene surfaces.
abstract: Ultra-high-molecular-weight polyethylene (UHMWPE) has a long history of use in medical devices, primarily for articulating surfaces due to its inherent low surface energy which limits tissue integration. To widen the applications of UHMWPE, the surface energy can be increased. The increase in surface energy would improve the adsorption of proteins and attachment of cells to allow tissue integration, thereby allowing UHMWPE to potentially be used for a wider range of implants. The attachment and function of human primary osteoblast-like (HOB) cells to surfaces of UHMWPE with various levels of incorporated surface oxygen have been investigated. The surface modification of the UHMWPE was produced by exposure to a UV/ozone treatment. The resulting surface chemistry was studied using X-ray photoelectron spectroscopy (XPS), and the topography and surface structure were probed by atomic force microscopy (AFM) and scanning electron microscopy (SEM), which showed an increase in surface oxygen from 11 to 26 atom % with no significant change to the surface topography. The absolute root mean square roughness of both untreated and UV/ozone-treated surfaces was within 350-450 nm, and the water contact angles decreased with increasing oxygen incorporation, i.e., showing an increase in surface hydrophilicity. Cell attachment and functionality were assessed over a 21 day period for each cell-surface combination studied; these were performed using SEM and the alamarBlue assay to study cell attachment and proliferation and energy-dispersive X-ray (EDX) analysis to confirm extracellular mineral deposits, and total protein assay to examine the intra- and extracellular protein expressed by the cells. HOB cells cultured for 21 days on the modified UHMWPE surfaces with 19 and 26 atom % oxygen incorporated showed significantly higher cell densities compared to cells cultured on tissue culture polystyrene (TCPS) from day 3 onward. This indicated that the cells attached and proliferated more readily on the UV/ozone-treated UHMWPE surfaces than on untreated UHMWPE and TCPS surfaces. Contact guidance of the cells was observed on the UHMWPE surfaces by both SEM and AFM. Scanning electron micrographs showed that the cells were confluent on the modified UHMWPE surfaces by day 10, which led to visible layering of the cells by day 21, an indicator of nodule formation. In vitro mineralization of the extracellular matrix expressed by the HOB cells on the modified UHMWPE surfaces was confirmed by SEM and EDX analysis; spherulite structures were observed near cell protrusions by day 21. EDX analysis confirmed the spherulites to contain calcium and phosphorus, the major constituents in calcium phosphate apatite, the mineral phase of bone. Overall cell attachment, functionality, and mineralization were found to be enhanced on the UV/ozone-modified UHMWPE surfaces, demonstrating the importance of optimizing the surface chemistry for primary HOB cells.
authors.affiliation: Materials & Biomaterials Research Centre, The Robert Gordon University, Aberdeen AB10 1FR, United Kingdom. spoulsson@gmail.com
authors.forename: Alexandra H C
authors.initials: AH
authors.lastname: Poulsson
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Mitchell
authors.forename: Marcus R
authors.initials: MR
authors.lastname: Davidson
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Johnstone
authors.forename: Neil
authors.initials: N
authors.lastname: Emmison
authors.forename: Robert H
authors.initials: RH
authors.lastname: Bradley
mesh_terms: D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods; D049109:Cell Proliferation; D002478:Cells, Cultured; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods; D008855:Microscopy, Electron, Scanning / Q000379:methods; D010006:Osteoblasts / Q000166:cytology*; D010100:Oxygen / Q000737:chemistry; D010126:Ozone; D020959:Polyethylene / Q000737:chemistry*; D011095:Polyethylenes / Q000737:chemistry; D012984:Software; D014466:Ultraviolet Rays; D014965:X-Rays
chemical_list: D011095:Polyethylenes; C111601:ultra-high molecular weight polyethylene; D010126:Ozone; D020959:Polyethylene; D010100:Oxygen