Penetration enhancement microscopy

In addition, data obtained from infrared, thermal, and fluorescence spectroscopic studies of the outermost layer of skin, stratum corneum (SC), and its components imply enhancer-improved permeation of solutes through the SC is associated with alterations involving the hydrocarbon chains of the SC lipid components. Data obtained from electron microscopy and x-ray diffraction reveals that the disordering of the lamellar packing is also an important mechanism for increased permeation of drugs induced by penetration enhancers (for a recent review, see Ref. 206). 

Infrared microscopic imaging provides the significant advantages of direct spatially resolved concentration and molecular structure information for sample constituents. Raman microscopy (not further discussed in this chapter) possesses the additional benefit of confocal acquisition of this information and a 10-fold increase in spatial resolution at the expense of reduced signal-to-noise ratios compared with IR. The interested reader is urged to check the seminal studies of the Puppels group in Rotterdam,38 0 as well as our own initial efforts in this direction.41 The current section describes the initial applications of IR microspectroscopic imaging to monitor the permeation and tissue distribution of the dermal penetration enhancer, DMSO, in porcine skin as well as to track the extent of permeation of phospholipid vesicles. 

In a more recent study, vesicle skin interactions were examined with confo-cal laser scanning microscopy (CLSM) [36], A large number of liposome compositions were examined. These studies revealed that the liposome constituents were only present in the outermost layers of the SC and that the constituents only acted as penetration enhancers, which is in contrast to the studies of Holland et al. Whether these differences in findings are due to another study design or to a difference in vesicle components is not clear. 

Different light microscopy techniques, such as autoradiography and quantitative fluorescence microscopy (QFM), have been used to examine the influence of penetration enhancers on percutaneous absorption. 

As previously discussed, electron, light, and confocal microscopy techniques may be used to visualize the position of electron-dense precipitates, radioactive substances, and fluorescent probes, respectively, in the sample tissue. However, none of these techniques possess the capability both to visualize and to selectively measure the flux of a molecule across the skin. SECM, however, permits the measurement and subsequent imaging of the local flux of an electroactive species across biological membranes. Scott et al. [3] used SECM to investigate the effect of pretreatment of the penetration enhancer sodium dodecyl sulfate (SDS), on the ion transport rate and transport pathways of Fe(CN) across hairless mouse skin. Increasing the time of SDS exposure from 10 min to 30 min increased the overall (porous and nonporous) transport of Fe(CN) by 17-fold. More specifically, the SDS-induced increase in Fe(CN)g transport was found to be associated with nonporous (i.e., intercellular) transport routes, while transport via porous routes was significantly reduced. The fraction of Fe(CN)g transport through pores, as measured by... 

There are many cases in which other techniques have been applied to biphasic systems in order to establish the nature of mixing. For example, fluorescence microscopy of DPPC monolayers containing 2% of a fluorescent probe have shown the coexistence of solid and fluid phases of DPPC at intermediate pressures (Weis, 1991). Similar results have been achieved with a variety of other phospholipids using the same technique (Vaz et al., 1989). The recent application of laser light scattering to this area (Street et al., unpublished data) has yet to produce any conclusive evidence, but the future for this particular technique is also promising. It also provides information about the viscoelastic properties of the monolayer and how these are affected by the inclusion of penetration enhancers. 

Saunders JCJ, Davis HJ, Coetzee L, et al. 1999. A Novel Skin Penetration Enhancer Evaluation by Membrane Diffusion and Confocal Microscopy. J Pharm Pharmaceut Sci 2(3) 99-107. http //www.ualberta.ca/ csps/JPPS2(3)/J.Saunders/microscopy.htm. 

The topic of ocular bioavailability of indomethacin administered in a sub-micrometer emulsion was al.so examined by the research group led by M, J. Alon.so at the University of Santiago de Compo-stela, Spain (32). These workers tested several carriers in vivo in rabbits nanopaiticles, nanocapsules, microparticles, and a submicrometer emulsion. The latter, prepared by the method of Yu et al. (33) with poloxamer 188 and soybean lecithin and containing 0.1% w/v indomethacin, had particles of diameter 0.21 0.02 im. The submicrometer systems (nanoparticles, nanocapsules, and emulsion) increased by more than three-fold the indomethacin concentration in the cornea, aqueous humor, and iris-ciliary body at 0.S and 1.0 h post instillation. Furthermore, an increased indomethacin bioavailability of 300% was observed in comparison with the value obtained for an aqueous commercial solution. Confocal laser scanning microscopy studies indicated (hat the submtcion particles penetrated into the comeal epithelium cells by endocyiosis (he audiors further suggested that the vehicle components (lecithin in the case of emulsions) may act as penetration enhancers or as endocytotic stimulators. 

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