Hemostatic Comparison of a Polysaccharide Powder and a Gelatin Powder.

Purpose/Aim: Powdered hemostats have been widely adopted for their ease-of-use; however, their efficacy has been limited resulting in applications restricted to low-level bleeds. This study investigates the use of bovine-derived gelatin particles (BGP) as a standalone hemostatic powder and compare BGP to commercially available microporous polysaccharide hemospheres (MPH). Materials and Methods: The powders were investigated for their hemostatic efficacy in a heparinized pre-clinical bleeding model limited to grade 1 and 2 bleeds on a validated intraoperative bleeding scale, which represents the accepted, clinical use of hemostatic powders. Results: At 10 minutes, the hemostatic success of lesions treated with BGP were 78% while MPH were 22%. The odds ratio for hemostatic success of BGP relative to MPH was 15.18 (95% CI: 7.37, 31.27). The 95% lower limit of the odds ratio was greater than 1. This indicates that BGP are superior to MPH (p < 0.001). The median time to hemostasis for BGP was 1.6 minutes and MPH was 14.5 minutes. The ratio for time to hemostasis of MPH relative to BGP was 9.23 (95% CI: 6.99, 12.19). This indicates that BGP achieve significantly faster time to hemostasis (p < 0.001). Conclusions: Characterization of tissue explant ultrastructure, particle size, and swelling revealed differences in the materials. BGP, in addition to absorbing fluid and concentrating clotting factors and platelets, integrate into the clot and stabilize the fibrin matrix. BGP have advantages over MPH in terms of speed and efficacy. BGP are a favorable biomaterial for further research that greatly improve the limited efficacy of powdered hemostats.

Production and analysis of a Bacillus subtilis biofilm comprised of vegetative cells and spores using a modified colony biofilm model.

Bacillus subtilis is a spore-forming soil bacterium that is capable of producing robust biofilms. Sporulation can occur in B. subtilis biofilms and it is possible that the spores embedded in the protective matrix could present a significant challenge to disinfecting agents or processes. This article describes a method for the growth and quantification of a reproducible B. subtilis ATCC 35021 biofilm comprised of vegetative cells and spores using a modified colony biofilm model. In this method, membranes were inoculated and incubated for a total of 8 days to promote biofilm formation and subsequent sporulation within the biofilm. Representative samples were taken over the course of the incubation period to evaluate the biofilms using enumerative, microscopic, and spectrometric methods. At various time points, the total numbers of cells and spores were quantified. A Congo red agar (CRA) method was utilized to detect the TasA matrix protein, a primary component of the B. subtilis biofilm matrix. The presence of TasA was also confirmed using mass spectrometry. The biofilm morphologies were correlated to the enumeration data with a variety of correlative imaging techniques: confocal microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). At the end of the incubation period, the biofilm contained >7 logs total colony forming units with spores comprising approximately 10% of the biofilm. The biofilm generated using this method allows researchers to use a new, more robust challenge for efficacy testing of chemical and physical antimicrobial treatments such as antibiotics, disinfectants, or heat.

Electric Field Controlled Self-Assembly of Hierarchically Ordered Membranes.

Self-assembly in the presence of external forces is an adaptive, directed organization of molecular components under nonequilibrium conditions. While forces may be generated as a result of spontaneous interactions among components of a system, intervention with external forces can significantly alter the final outcome of self-assembly. Superimposing these intrinsic and extrinsic forces provides greater degrees of freedom to control the structure and function of self-assembling materials. In this work we investigate the role of electric fields during the dynamic self-assembly of a negatively charged polyelectrolyte and a positively charged peptide amphiphile in water leading to the formation of an ordered membrane. In the absence of electric fields, contact between the two solutions of oppositely charged molecules triggers the growth of closed membranes with vertically oriented fibrils that encapsulate the polyelectrolyte solution. This process of self-assembly is intrinsically driven by excess osmotic pressure of counterions, and the electric field is found to modify the kinetics of membrane formation, and also its morphology and properties. Depending on the strength and orientation of the field we observe a significant increase or decrease of up to nearly 100% in membrane thickness, as well as the controlled rotation of nanofiber growth direction by 90 degrees, resulting in a significant increase in mechanical stiffness. These results suggest the possibility of using electric fields to control structure in self-assembly processes involving diffusion of oppositely charged molecules.

A self-assembly pathway to aligned monodomain gels.

Aggregates of charged amphiphilic molecules have been found to access a structure at elevated temperature that templates alignment of supramolecular fibrils over macroscopic scales. The thermal pathway leads to a lamellar plaque structure with fibrous texture that breaks on cooling into large arrays of aligned nanoscale fibres and forms a strongly birefringent liquid. By manually dragging this liquid crystal from a pipette onto salty media, it is possible to extend this alignment over centimetres in noodle-shaped viscoelastic strings. Using this approach, the solution of supramolecular filaments can be mixed with cells at physiological temperatures to form monodomain gels of aligned cells and filaments. The nature of the self-assembly process and its biocompatibility would allow formation of cellular wires in situ that have any length and customized peptide compositions for use in biological applications.