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The settlement and release of Ulva spores from chemically modified, micro-engineered surface topographies have been investigated using poly(dimethyl siloxane) elastomers (PDMSe) with varying additions of non-network forming poly(dimethyl siloxane) based oils. The topographic features were based on 5 microns wide pillars or ridges separated by 5, 10, or 20 microns wide channels. Pattern depths were 5 or 1.5 microns. Swimming spores showed no marked difference in settlement on smooth surfaces covered with excess PDMS oils. However, incorporation of oils significantly reduced settlement density on many of the surfaces with topographic features, in particular, the 5 microns wide and deep channels. Previous results, confirmed here, demonstrate preferences by the spores to settle in channels and against pillars with spatial dimensions of 5 microns, 10 microns and 20 microns. The combination of lubricity and pillars significantly reduced the number of attached spores compared to the control, smooth, unmodified PDMSe surfaces when exposed to turbulent flow in a flow channel. The results are discussed in relation to the energy needs for spores to adhere to various surface features and the concepts of ultrahydrophobic surfaces. A factorial, multi-level experimental design was analyzed and a 2nd order polynomial model was regressed for statistically significant effects and interactions to determine the magnitude and direction of influence on the spore density measurements between factor levels.

We examined how variations in elastic modulus, surface chemistry and the height and spacing of micro-ridges interact and effect endothelial cell (EC) alignment. Specifically, we employed independent control of the surface properties in order to elucidate the relative importance of each factor. Polydimethylsiloxane elastomer (PDMSe) was fabricated with 1.5 or 5 microm tall, 5 microm spaced and 5, 10, or 20 microm wide ridge microtopographies. Elastic modulus was varied from 0.3, 1.0, 1.4, and 2.3 MPa by controlling oligomeric additives and crosslink density. Surface chemistry was left untreated, argon plasma treated, coated with fibronectin (Fn) or patterned with Fn tracks on flat PDMSe or the tops of micro-ridges. Primary porcine vascular ECs were cultured on the PDMSe substrates and nuclear form factor (NFF) was used to determine cell orientation relative to surface microtopography. Experimental results showed that microtopographical variation strongly altered EC alignment on Fn coated surfaces, but not on plasma treated surfaces. Interestingly, similar alignment was achieved with different orientation cues, either micropatterned chemistry (2D) or microtopography (3D). In total, the effect of varying one of the experimental parameters depended strongly on the state of the others, highlighting the need for multi-factor analysis of surface properties for applications where cells and tissue will contact synthetic materials.

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An antigen-binding fragment (Fab) from a murine monoclonal antibody (4-4-20) with high affinity for fluorescein was cocrystallized with ligand in polyethylene glycol (PEG) and 2-methyl-2,4-pentanediol (MPD) in forms suitable for X-ray analyses. In MPD the affinity of the intact antibody for fluorescein was 300 times lower than the value (3.4 x 10(10) M-1) obtained in aqueous buffers. This decreased affinity was manifested by the partial release of bound fluorescein when MPD was added to solutions of liganded Fab during crystallization trials. In PEG, the ligand remained firmly bound to the protein. The liganded Fab crystallized in the monoclinic space group P2(1) in PEG, with a = 58.6, b = 97.2, c = 44.5 A and beta = 95.2 degrees. In MPD the space group was triclinic P1, with a = 58.3, b = 43.4, c = 42.3 A, alpha = 83.9 degrees, beta = 87.6 degrees, and gamma = 84.5 degrees. X-ray diffraction data were collected for both forms to 2.5-A resolution. Surprisingly, the triclinic form of the liganed antifluorescyl Fab had the same space group, closely similar cell dimensions, and practically the same orientation in the unit cell as an unliganded Fab (BV04-01) with activity against single-stranded DNA.