Charge stabilized crystalline colloidal arrays as templates for fabrication of non-close-packed inverted photonic crystals.

We developed a straightforward method to form non-close-packed highly ordered fcc direct and inverse opal silica photonic crystals. We utilize an electrostatically self assembled crystalline colloidal array (CCA) template formed by monodisperse, highly charged polystyrene particles. We then polymerize a hydrogel around the CCA (PCCA) and condense silica to form a highly ordered silica impregnated (siPCCA) photonic crystal. Heating at 450 degrees C removes the organic polymer leaving a silica inverse opal structure. By altering the colloidal particle concentration we independently control the particle spacing and the wall thickness of the inverse opal photonic crystals. This allows us to control the optical dielectric constant modulation in order to optimize the diffraction; the dielectric constant modulation is controlled independently of the photonic crystal periodicity. These fcc photonic crystals are better ordered than typical close-packed photonic crystals because their self assembly utilizes soft electrostatic repulsive potentials. We show that colloidal particle size and charge polydispersity has modest impact on ordering, in contrast to that for close-packed crystals.

Fast responsive crystalline colloidal array photonic crystal glucose sensors.

We developed new photonic crystal polymerized crystalline colloidal array (PCCA) glucose sensing materials, which operate on the basis of formation of cross-links in the hydrogel. These materials are composed of hydrogels that embed an array of approximately 100-nm-diameter monodisperse polystyrene colloids that Bragg diffract light in the visible spectral region. The hydrogels change volume as the glucose concentration varies. This changes the lattice spacing, which changes the wavelength of the diffracted light. In contrast to our previous glucose sensing photonic crystal materials, we no longer require Na+ chelating agents. These photonic crystal materials are being designed for use in glucose sensing contact lens for people with diabetes mellitus. We describe methods to speed up the response kinetics of these PCCA sensing materials. Rapid-response kinetics is achieved by controlling the elasticity and the hydrophilic-hydrophobic balance of the hydrogel system. A more hydrophobic hydrogel composition is obtained by copolymerizing n-hexylacrylate into an acrylamide-bisacrylamide hydrogel. The response rate significantly increases to where it fully responds within 90 s to the average glucose concentrations found in blood (5 mM) and within 300 s to the average glucose concentrations found in tear fluid (0.15 mM). We find unusual temperature-dependent kinetics, which derive from glucose mutarotation in solution. It is shown that alpha-d-glucose is the glucose anomer binding to the boronic acid derivative. Care must be taken in any glucose determination to ensure that the glucose mutarotation equilibrium has been established. We have demonstrated that the sensor is responsive to approximately 0.15 mM glucose concentrations in artificial tear fluid solution.

Liposome transduction into cells enhanced by haptotactic peptides (Haptides) homologous to fibrinogen C-termini.

Haptides are 19-21mer cell-binding peptides equivalent to sequences on the C-termini of fibrinogen beta chain (Cbeta), gamma chain (preCgamma) and the extended alphaE chain of fibrinogen (CalphaE). In solution, Haptides accumulated in cells by non-saturable kinetics [Exp. Cell Res. 287 (2003) 116]. This study describes Haptide interactions with liposomes and Haptide-mediated liposome uptake by cells. Haptides became incorporated into negatively charged liposomes, changing their zeta potential. Atomic force microscopy and particle sizing by light scattering showed that the liposomes dissolved Haptide nanoparticles and absorbed them from solution. Pre-mixing fluorescent rhodamine-containing liposomes or "stealth" doxorubicin (DOX)-containing liposomes (Doxil) with Cbeta, preCgamma or to a lesser degree CalphaE, significantly enhanced their uptake by fibroblasts and endothelial cells. Confocal microscopy showed Haptide-induced liposome uptake saturated above approximately 40 microM Haptide. Cytotoxicity tests with lower concentrations of Doxil liposomes indicated that premixing with approximately 40 microM Cbeta or preCgamma increased their toxicity by one order of magnitude. It was evident that the liposomes complexed with an amphiphilic Haptide are transduced through cell membranes, probably by a non-receptor-mediated process. These results suggest that Cbeta or pre-Cgamma could be employed to augment the cellular uptake of drugs in liposomal formulations.

Fibrinogen C-terminal peptidic sequences (Haptides) modulate fibrin polymerization.

We previously described synthetic peptides of 19-21 amino acid residues, homologous to the C-termini of fibrinogen Fib(340) and Fib(420), from the beta-chain (Cbeta), the extended alphaE chain (CalphaE) and near the end of the gamma-chain (preCgamma) which elicited attachment (haptotactic) responses from mesenchymal cells. We named these haptotactic peptides -Haptides. The effects of Haptides on fibrin clot formation was evaluated and their possible effects on platelet aggregation was examined. The Haptides Cbeta,CalphaE and preCgamma, (2-10 micro M) increased fibrin clot turbidity and also decreased thrombin-induced clotting time. Higher concentrations (>120 micro M of Cbeta or preCgamma) induced fibrinogen precipitation even without thrombin. These precipi-tates exhibited different ultrastructure from thrombin-induced fibrin by scanning and transmission microscopy. C-terminal peptides of the other fibrinogen chains exerted no such effects. Sepharose beads covalently coated with either whole fibrinogen or Haptides (SB-Fib or SB-Haptide) highly adsorbed free (FITC) Haptides. In aqueous solution, Haptides formed nano-par-ticles with average size of approximately 150 nm in diameter. We suggest that such positively charged aggregates could serve to nucleate and accelerate fibrin gel formation. These results also indicate that Cbeta and preCgamma sequences within fibrin(ogen) participate in the docking and condensation of fibrin(ogen) during its assembly into a fibrin clot. By contrast, Haptides up to 100 micro M did not bind to platelets, and had no effect on platelet aggregation. Our findings highlight the roles of the C-terminal sequences of the beta and gamma chains in fibrin(ogen) polymerization as well as in cell attachment.