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The influence of the ultrafine crystallinity of commercial purity grade 2 (as-received) titanium and titanium modified by equal channel angular pressing (modified titanium) on bacterial attachment was studied. A topographic profile analysis of the surface of the modified titanium revealed a complex morphology of the surface. Its prominent micro- and nano-scale features were 100-200-nm-scale undulations with 10-15 microm spacing. The undulating surfaces were nano-smooth, with height variations not exceeding 5-10 nm. These surface topography characteristics were distinctly different from those of the as-received samples, where broad valleys (up to 40-60 microm) were detected, whose inner surfaces exhibited asperities approximately 100 nm in height spaced at 1-2 microm. It was found that each of the three bacteria strains used in this study as adsorbates, viz. Staphylococcus aureus CIP 68.5, Pseudomonas aeruginosa ATCC 9025 and Escherichia coli K12, responded differently to the two types of titanium surfaces. Extreme grain refinement by ECAP resulted in substantially increased numbers of cells attached to the surface compared to as-received titanium. This enhanced degree of attachment was accompanied with an increased level of extracellular polymeric substances (EPS) production by the bacteria.

This work is part of a general effort to demonstrate the effect of the bulk microstructure of titanium as a model bone implant material on viability of osteoblasts (bone-forming cells). The objective of this work was to study the proliferation of preosteoblastic MC3T3-E1 cells extracted from mice embryos on commercial purity titanium substrates. Two distinct states of titanium were considered: as-received material with an average grain size of 4.5 microm and that processed by equal channel angular pressing (ECAP), with an average grain size of 200 nm. We report the first results of an in vitro study into the effect of this extreme grain refinement on viability and proliferation of MC3T3-E1 cells. By means of MTT assays it was demonstrated that ECAP processing of titanium enhances MC3T3-E1 culture proliferation in a spectacular way. This finding suggests that bone implants made from ECAP processed titanium may promote bone tissue growth.

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