Cellular attachment and spatial control of cells using micro-patterned ultra-violet/Ozone treatment in serum enriched media

Abstract

Ultra-violet Ozone (UVO) modified polystyrene (PS) surfaces were analyzed by X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), contact angle (CA), optical microscopy (OM) and cell culture experiments. UV/Ozone treatment up to 900 s was used to increase the surface oxygen concentration of PS surfaces from 0% to ∼35% (unwashed) and 0% to ∼27% (washed). The observed differences in oxygen concentration, between washed and unwashed surfaces, have been previously attributed to the removal of low molecular weight debris produced in this treatment process. Surface roughness ( R q) is known to affect cellular attachment and proliferation. AFM studies of the UV/Ozone treated PS surfaces show the surface roughness is an order of magnitude less than that expected to cause an effect. UV/Ozone treatment of PS showed a marked change in CA which decreased to ∼60° after 900 s treatment. The increased attachment and proliferation of Chinese hamster ovarian (CHO) and mouse embryo 3T3-L1 (3T3) cells on the treated surfaces compared to untreated PS were found to correlate strongly with the increase in surface oxygen concentration. Surface chemical oxidation patterns on the PS were produced using a simple masking technique and a short UV/Ozone treatment time, typically 20–45 s. The chemical patterns on PS were visualized by water condensation and the spatially selective attachment of CHO and 3T3-L1 cells cultured with 10% (v/v) serum. This paper describes an easily reproducible, one step technique to produce a well-defined, chemically heterogeneous surface with a cellular resolution using UV/Ozone modification. By using a variety of cell types, that require different media conditions, we have been able to expand the potential applications of this procedure.