Comparison of the transdermal delivery of estradiol from two gel formulations.

OBJECTIVE: Conventional oral oestrogen replacement therapy can relieve postmenopausal symptoms but is associated with undesirable side-effects which can be minimised by avoiding the fluctuating hormonal blood levels resulting from oral therapy and eliminating hepatic first-pass metabolism by the use of the transdermal route. The two commercially available transdermal gel formulations differ in composition and application recommendations. Sandrena Gel contains 0.1% (w/w) and Oestrogel 0.06% (w/w) estradiol and recommended dosages are 0.5-1.5 g over 200-400 cm2 (Sandrena Gel) and 2.5 g gel over 720 cm2 (Oestrogel). In transdermal therapy the formulation composition may have a significant effect on drug delivery and we have therefore compared the permeation of estradiol from these formulations across human skin in vitro. METHODS: The in vitro percutaneous penetration of estradiol from the formulations through epidermal membranes prepared from excised female human thing skin was assessed over a 24 h period using static type Franz diffusion cells. RESULTS: Permeation of the active was similar from each formulation representing (at 24 h) 18.2 +/- 3.5% of the applied dose from Sandrena Gel and 17.4 +/- 4.8% of the applied dose from Oestrogel. These percentages equate to cumulative skin permeations of 0.65 +/- 0.15 microgram/cm2 and 0.45 +/- 0.15 microgram/cm2 respectively. CONCLUSION: The results suggest that the two formulations are bioequivalent at the recommended dose levels.

Molecular structural changes in human fetal tissue during the early stages of embryogenesis.

Low-angle synchrotron X-ray diffraction studies of human fetal extensor tendon and skin collagen, centred in time about the period of first major movement by the fetus, indicate that alignment of the tendon collagen fibrils occurs about this time. At this stage there appears to be no detectable structural difference between tendon and dermis. By three weeks post-partum, marked differences between these tissues can be detected. The distribution of the intermediate filament diameters for all fetal tendons investigated was unimodal (mean 41.2 +/- 0.4 nm) in contrast with that for post-partum tendon which is multimodal. Equatorial periodicities of 353 +/- 3 nm and 32.1 +/- 0.1 nm, consistent with the presence of type IV collagen, were obtained from all fetal samples examined. Neither of these periodicities were observed in post-partum normal tendon and only the larger were observed in post-partum normal skin. The consistency of the results suggest that low-angle X-ray diffraction could be used for the identification of fetal-like tissues found in pathological tissues.

Percutaneous penetration of N-nitroso-N-methyldodecylamine through human skin in vitro: application from cosmetic vehicles.

The human skin penetration of N-nitroso-N-methyldodecylamine (NDOMA) from isopropyl myristate (IPM) and two vehicles representative of cosmetic/personal care formulations was determined in vitro. When applied as an infinite dose in IPM (1 microgram/microliter) the average total absorption over 48 hr was 0.10 +/- 0.01% of the applied dose (all data are expressed as means +/- SE). When applied as a finite dose in a representative oil-in-water emulsion formulation the average total absorption over 48 hr was 4.66 +/- 0.76% of the applied dose. When applied as a finite dose in a representative shampoo formulation for 10 min, followed by rinsing (to represent in-use exposure conditions), the average total absorption over 48 hr was 0.75 +/- 0.17% of the applied dose. Approximately 72% of the NDOMA in the applied shampoo formulation was removed by rinsing. The overall data indicated that NDOMA could penetrate the skin but that penetration was low. The rate and extent of absorption, however, could be affected by differences in the vehicle of application, time of exposure and whether the formulation is (and the conditions are designed to mimic) a rinse-off or leave-on product.

Changes in the molecular structure of hair in insulin-dependent diabetes.

Low-angle synchrotron X-ray diffraction has revealed clear and consistent changes in the molecular structure of alpha-keratin of hair in insulin-dependent diabetes (IDDM) both for human IDDM subjects and for baboons with streptozocin induced diabetes. These changes in both meridional and equatorial intensity distributions are fully explained by a newly developed hexagonally packed model for keratin which locates the modification produced in hair in IDDM in the labile structure of the matrix at established intermediate filament linkage sites. The nature of the extracellular bonding suggests that the change is endogenous, occurring via the blood during the aggregation of the IFs in the follicle. The reproducibility of these changes indicate that hair may represent an easily accessible tissue for the study of how hyperglycaemia can modify extracellular matrix materials which lead to diabetic complications.

Percutaneous penetration of octyl salicylate from representative sunscreen formulations through human skin in vitro.

The human skin penetration of [14C]octyl salicylate from two representative sunscreen vehicles was determined in vitro. 3H-sucrose was incorporated into all formulations and provided a marker for membrane integrity. When applied as a finite dose in an oil-in-water emulsion vehicle containing 5% (w/w) octyl salicylate, the average total absorption of 14C over 48 hr was 0.65+/-0.16% of the applied dose (representing a total amount permeated of 1.58+/-0.36 microg/cm2). When applied as an infinite dose in the oil-in-water emulsion vehicle the average total absorption of 14C over 48 hr was 0.47+/-0.22% of the applied dose (representing a total amount permeated of 27.54+/-13.91 microg/cm2). When applied as a finite dose in a representative hydroalcoholic formulation containing 5% (w/w) octyl salicylate, the average total absorption of 14C over 48 hr was 0.59+/-0.09% of the applied dose (representing a total amount permeated of 1.58+/-0.25 microg/cm2). When applied as an infinite dose in the hydroalcoholic formulation the average total absorption of 14C over 48 hr was 0.23+/-0.05% of the applied dose (representing a total amount permeated of 11.28+/-2.55 microg/cm2). The penetration of [14C]salicylic acid [applied at a concentration of 2.7% (w/w), in the oil-in-water emulsion] was also determined. When applied as a finite dose the average total absorption of 14C over 48 hr was 1.14+/-0.23% of the applied dose (representing a total amount permeated of 1.65+/-0.39 microg/cm2). These results suggest that the in vitro human skin permeation of octyl salicylate is relatively low. The amounts of octyl salicylate and salicylic acid permeated when applied in similar vehicles were remarkably similar over 48 hr (1.58 microg/cm2 and 1.65 microg/cm2, respectively). This suggests the possibility that the 14C label appearing in the receptor fluid may, in both cases, represent salicylic acid. If this is the case, then it is possible that the amount of octyl salicylate permeating through the skin is much less than that suggested by the data obtained here. This supposition is, however, entirely speculative and has yet to be confirmed experimentally.

The intermediate filament structure of human hair.

X-ray diffraction studies of hard alpha-keratin have led to a proposed model for the lateral arrangement of molecules within the keratin fibrils of tissues such as hair, nail and claw. Using low-angle synchrotron radiation to examine human scalp hair we have obtained discrete equatorial diffraction maxima which have not been reported previously. These reflections can be divided into three subsets. The first of these reveals the information that the hair fibres consist basically of cylindrical fibrils arranged in a disordered lattice. The mean diameters of these cylinders have been determined, together with their average separation. The diameters of the protofibrils have been determined from the second set. The third set, a set of diffuse arcs, index onto a spacing which is characteristic of the disordered components of the matrix.

Intermediate filament structure of alpha-keratin in baboon hair.

High-quality photographic recording of X-ray diffraction data from the hard alpha-keratins has mainly been limited to that from porcupine quill. Earlier diffraction patterns of hair have shown very few reflections. In the present study, extensive sets of high-quality data have been obtained using a synchrotron source and an image plate detector, revealing information about the internal structure, both axial and lateral. The diffraction patterns of hair from six different baboons have been examined. The meridional axial diffraction data reveal evidence of the superposition of three and possibly four separate lattices. Despite considerable overlapping, sufficient reflections unique to each lattice have been observed. These lattices comprise an infinite one of 46.76 nm, together with finite lattices of 19.67, 27.20 and 7.27 nm. These patterns reveal information regarding the fibrillar structure of the hair fibres, namely that it consists basically of cylindrical fibrils arranged in a disordered lattice. The mean diameters of these cylinders have been determined, together with their average separation. There is also an indication of the presence of another set of cylinders of smaller diameter, possibly comprising the microfibrils of keratin.

Percutaneous penetration of dimethylnitrosamine through human skin in vitro: application from cosmetic vehicles.

Human skin penetration of N-dimethylnitrosamine (DMN) from three vehicles has been determined in vitro. When applied as an infinite dose in isopropyl myristate (IPM, 1 microgram/microliter) the average total absorption over 48 hr was 2.6 +/- 1.2% of the applied dose (all data presented are expressed as means +/- standard errors). When applied as a finite dose in a representative oil-in-water emulsion vehicle the average total absorption over 48 hr was 4.0 +/- 0.3% of the applied dose. When applied as a finite dose in a representative shampoo vehicle for 10 min followed by rinsing (i.e. to represent in-use exposure conditions) the average total absorption over 48 hr was 1.1 +/- 0.1% of the applied dose. Approximately 72% of the DMN in the applied shampoo vehicle was removed by rinsing. There was considerable evaporative loss of DMN from the IPM and oil-in-water emulsion vehicles, such that absorption was complete within 3 hr of application. The overall data indicate that DMN can penetrate the skin rapidly but that in practice the amount actually available for penetration is significantly reduced by high permeant volatility. In contrast, application of N-nitrosodiethanolamine (NDELA) at a concentration of 1 microgram/microliter as an infinite dose generated an average total absorption over 48 hr of 23.6 +/- 6.4%, representing a total flux of 103.9 +/- 28.4 micrograms/cm2. In the case of NDELA, no evaporative loss was evident.

Two very long periodicities in collagen.

A low-angle X-ray diffraction study of skin and breast-duct collagen, using synchrotron radiation, has disclosed an equatorial periodicity, common to all specimens, of about 350 nm. Tendons did not show any such periodicity, indicating that this phenomenon is restricted to two-dimensional distributions. In addition, shorter equatorial periodicities were also in evidence, which were in most instances different for different specimens, occurring in the range of 40 to 140 nm. Electron micrographs suggest the possibility that the first periodicity could be linked with blocks of the collagen molecule, which has a length of about 300 nm. The shorter periodicities give the tissue specific interfibrillar lateral packing of the individual collagen microfibrils.

The d-spacing of collagen from mitral heart valves changes with ageing, but not with collagen type III content.

A low angle X-ray diffraction study of normal and myxomatous human heart valves has been carried out using a Synchrotron radiation source. This study has shown a slight progressive decrease in the d-spacing of the collagen with age. Despite suggestions reported in the literature, no significant relative change was observed in the d-spacing with the increase in the percentage of collagen type III from 24% in normal to 31% in myxomatous heart valves.

Use of X-ray diffraction in study of human diabetic and aging collagen.

Extensive investigations of the solubility and fluorescence of collagen fibers in diabetes have revealed that there are significant changes in their physical properties. These changes are associated with increased cross-link formation. We used X-ray diffraction to study these changes in human extensor tendons at a molecular level in relation to both aging and diabetes. Our results indicate that diabetes induces significant alterations in the ultrastructure of collagen in the lateral packing of the molecules and the axial structure of the specimen. These changes can be induced in normal tendon by incubation in ribose and glucose-6-phosphate but are different from those associated with the normal process of aging.

Location and effect of procaine on lecithin/cholesterol membranes using X-ray diffraction methods.

X-ray diffraction studies were made on lecithin/cholesterol multilayers with very high water content and containing the local anaesthetic procaine. Narrow-angle diffraction experiments show that the procaine molecules are located with the uncharged aromatic amine group approx. 10 A from the centre of the bilayer. The polar tertiary amine group of these molecules is almost certainly located in the polar head-group region of the membrane. Wide-angle diffraction experiments show that the incorporation of procaine molecules into such lipid membranes produces an approx. 30% increase in the spread of acyl chain separation, although the average spacing between the chains is slightly reduced.