Filamentous Escherichia coli cells swimming in tapered microcapillaries.

This study analyzed the swimming characteristics of filamentous Escherichia coli cells inside tapered capillaries with a diameter decreasing from 700 µm to 4 µm and a mean body length of 27.8 µm ± 11.9 µm. Cells that were pre-oriented towards the narrower diameter section of the tapered capillary swam with high directional persistence, following conical-helix trajectories along the capillary wall. The confinement of the tapered capillary significantly diminished the mean swimming speed of filamentous cells when compared to their unrestricted mean swimming speed. The cell body rotation of individual filamentous bacteria decreased along the tapered direction, likely due to increased steric interactions with the capillary wall. Filamentous cells that swam under imposed flow rates ranging from 0.2 µl/min to 0.8 µl/min showed positive rheotaxis inside the 150 µm-350 µm diameter region of the tapered capillary. Depending on the imposed flow rate, none of the bacteria could advance beyond a critical diameter in the tapered capillary. This critical diameter is likely to be the position of the maximum shear rate they can tolerate without being flushed away. This work showed experimental evidence of how a simple flow constriction such as a tapered tube forms a hydrodynamic barrier that can deter the advance of bacterial rheotaxis.

Improved dermal delivery of FITC-BSA using a combination of passive and active methods.

This work presents results on the in vitro penetration of a model macromolecule [fluorescein isothiocyanate-labeled bovine serum albumin (FITC-BSA)] through porcine skin, mediated with a microneedle skinroller (200-µm-length needle) and different novel formulations. After perforating the porcine skin with a microneedle skinroller, the efficiency of delivering FITC-BSA via different novel formulations was evaluated and compared. Formulations, including l-α-phosphatidylcholine (PC) liposomes, double emulsions, and double-encapsulation formulations were used. High-resolution cryo-scanning electron microscopy was used to visualize surface morphology and cross-section of perforated porcine skin. By the use of confocal microscopy, the penetration pathway and penetration depth of FITC-BSA through the perforated porcine skin under different formulations were analyzed. FITC-BSA was extracted from stratum corneum and viable skin, and analyzed by fluorimetry, indicating that there is no significant difference in the amount of FITC-BSA delivered to viable skin by PC-liposome suspension (12.90 ± 1.25 µg/cm(2)) versus double-encapsulation formulations (10.47 ± 0.80 µg/cm(2)); however, both formulations showed a significant increase as compared with an aqueous solution of FITC-BSA. In this work, double-encapsulation formulations were used in dermal delivery for the first time and combined with microneedle skinroller treatment, the results showed a high efficiency in delivering macromolecules.

Oil-frozen W1/O/W2 double emulsions for dermal biomacromolecular delivery containing ethanol as chemical penetration enhancer.

Oil-frozen water-in-oil-in-water (W1/O/W2) double emulsions (DE) containing ethanol up to 40% (w/v) in the external aqueous W2 phase exhibited external coalescence upon thawing of the oil phase, releasing up to 85% of the encapsulated protein of the internal aqueous phase. These emulsions were studied in vitro as potential dermal macromolecular delivery formulations, achieving fluorescein isothiocyanate-labeled bovine serum albumin (FITC-BSA) penetration of up to 86 µm into porcine skin, reaching the viable epidermis where the immunocompetent Langerhans cells are located. Enzyme-linked immunosorbent assay was performed to observe the effect of the emulsification process and ethanol content on the ability of BSA to form antigen-antibody complexes; results indicated that ethanol content and the emulsification process did not diminish the BSA-antibody complex formation when compared with a BSA standard aqueous solution. Therefore, it is shown that oil-frozen W1/O/W2 DE, with penetration-enhancing ethanol in the W2 phase, can potentially be used for cutaneous vaccine delivery formulations.