Spatial and orientation control of cylindrical nanostructures in ABA triblock copolymer thin films by raster solvent vapor annealing.

We present a spatially resolved approach for the solvent vapor annealing (SVA) of block copolymer thin films that permits the facile and relatively rapid manipulation of nanoscale ordering and nanostructure orientation. In our method, a localized (point) SVA zone is created through the use of a vapor delivery nozzle. This point annealing zone can be rastered across the thin film using a motorized stage to control the local nanoscale structure and orientation in a cylinder-forming ABA triblock copolymer thin film. At moderate rastering speeds (∼100 µm/s) (i.e., relatively modest annealing time at a given point), the film displayed ordered cylindrical nanostructures with the cylinders oriented parallel to the substrate surface. As the rastering speed was decreased (∼10 µm/s), the morphology transformed into a surface nanostructure indicative of cylinders oriented perpendicular to the substrate surface. These perpendicular cylinder orientations also were created by rastering multiple times over the same region, and this effect was found when rastering in either retrace (overlapping) or crossed-path (orthogonal) geometries. Similar trends in nanostructure orientation and ordering were obtained from various nozzle diameters by accounting for differences in solvent flux and annealing time, illustrating the universality of this approach. Finally, we note that our "stylus-based" raster solvent vapor annealing technique allows a given point to be solvent annealed approximately 2 orders of magnitude faster than conventional "bell jar" solvent vapor annealing.

Systematic study on the effect of solvent removal rate on the morphology of solvent vapor annealed ABA triblock copolymer thin films.

Nanoscale self-assembly of block copolymer thin films has garnered significant research interest for nanotemplate design and membrane applications. To fulfill these roles, control of thin film morphology and orientation is critical. Solvent vapor annealing (SVA) treatments can be used to kinetically trap morphologies in thin films not achievable by traditional thermal treatments, but many variables affect the outcome of SVA, including solvent choice, total solvent concentration/swollen film thickness, and solvent removal rate. In this work, we systematically examined the effect of solvent removal rate on the final thin film morphology of a cylinder-forming ABA triblock copolymer. By kinetically trapping the film morphologies at key points during the solvent removal process and then using successive ultraviolet ozone (UVO) etching steps followed by atomic force microscopy (AFM) imaging to examine the through-film morphologies of the films, we determined that the mechanism for cylinder reorientation from substrate-parallel to substrate-perpendicular involved the propagation of changes at the free surface through the film toward the substrate as a front. The degree of reorientation increased with successively slower solvent removal rates. Furthermore, the AFM/UVO etching scheme permitted facile real-space analysis of the thin film internal structure in comparison to cross-sectional transmission electron microscopy.

Gradient solvent vapor annealing of block copolymer thin films using a microfluidic mixing device.

Solvent vapor annealing (SVA) with solvent mixtures is a promising approach for controlling block copolymer thin film self-assembly. In this work, we present the design and fabrication of a solvent-resistant microfluidic mixing device to produce discrete SVA gradients in solvent composition and/or total solvent concentration. Using this device, we identified solvent composition dependent morphology transformations in poly(styrene-b-isoprene-b-styrene) films. This device enables faster and more robust exploration of SVA parameter space, providing insight into self-assembly phenomena.

RhoA/Rho-kinase suppresses endothelial nitric oxide synthase in the penis: a mechanism for diabetes-associated erectile dysfunction.

Significant impairment in endothelial-derived nitric oxide is present in the diabetic corpus cavernosum. RhoA/Rho-kinase may suppress endothelial nitric oxide synthase (eNOS). Here, we tested the hypothesis that RhoA/Rho-kinase contributes to diabetes-related erectile dysfunction and down-regulation of eNOS in the streptozotocin (STZ)-diabetic rat penis. Colocalization of Rho-kinase and eNOS protein was present in the endothelium of the corpus cavernosum. RhoA/Rho-kinase protein abundance and MYPT-1 phosphorylation at Thr-696 were elevated in the STZ-diabetic rat penis. In addition, eNOS protein expression, cavernosal constitutive NOS activity, and cGMP levels were reduced in the STZ-diabetic penis. To assess the functional role of RhoA/Rho-kinase in the penis, we evaluated the effects of an adeno-associated virus encoding the dominant-negative RhoA mutant (AAVTCMV19NRhoA) on RhoA/Rho-kinase and eNOS and erectile function in vivo in the STZ-diabetic rat. STZ-diabetic rats transfected with AAVCMVT19NRhoA had a reduction in RhoA/Rho-kinase and MYPT-1 phosphorylation at a time when cavernosal eNOS protein, constitutive NOS activity, and cGMP levels were restored to levels found in the control rats. There was a significant decrease in erectile response to cavernosal nerve stimulation in the STZ-diabetic rat. AAVT19NRhoA gene transfer improved erectile responses in the STZ-diabetic rat to values similar to control. These data demonstrate a previously undescribed mechanism for the down-regulation of penile eNOS in diabetes mediated by activation of the RhoA/Rho-kinase pathway. Importantly, these data imply that inhibition of RhoA/Rho-kinase improves eNOS protein content and activity thus restoring erectile function in diabetes.