William L. Pickens, Ronald R. Warner, Ying L. Boissy, Raymond E

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Characterization of Vernix Caseosa: Water Content, Morphology, and Elemental Analysis  William L. Pickens, Ronald R. Warner, Ying L. Boissy, Raymond E. Boissy, Steven B. Hoath  Journal of Investigative Dermatology  Volume 115, Issue 5, Pages 875-881 (November 2000) DOI: 10.1046/j.1523-1747.2000.00134.x Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Dry weight: wet weight ratios of vernix caseosa and standard topical creams used in the newborn nursery. The data indicate that vernix, obtained at birth from term infants, is approximately 80% volatile. Seventeen percent of Eucerin was volatile with negligible volatility observed in the other preparations. All samples were dried to constant weight over a 1 wk period. The dry weight to wet weight ratios of vernix and Eucerin were each statistically different from the other preparations using one-way ANOVA and Bonferroni t test. Results are reported as mean ± SD, *p < 0.05. Journal of Investigative Dermatology 2000 115, 875-881DOI: (10.1046/j.1523-1747.2000.00134.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Dehydration kinetics of vernix caseosa and standard topical creams following hydration in normal saline. (a) Vernix and standard barrier creams were hydrated in normal saline over a 68 h period. Petrolatum and Aquaphor exhibited relatively small decreases in weight following hydration. Eucerin demonstrated a slight increase in weight, whereas, on average, vernix caseosa weight did not change. (b) Following hydration, the specimens were allowed to desiccate under ambient conditions. The rate of desiccation over 8 d is shown. All data are reported as mean ± SD. Journal of Investigative Dermatology 2000 115, 875-881DOI: (10.1046/j.1523-1747.2000.00134.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Rehydration kinetics of desiccated vernix caseosa in either normal saline or deionized water. Following complete desiccation, vernix was rehydrated over a period of 5 d in either deionized water or normal saline. Rehydration rates and the percentage of rehydration were determined by comparison with the original specimen weight. Data are reported as mean ± SD. Journal of Investigative Dermatology 2000 115, 875-881DOI: (10.1046/j.1523-1747.2000.00134.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Low magnification and phase contrast microscopy of vernix caseosa. These photomicrographs depict the dual nature of vernix caseosa: (a) A specimen of freshly harvested vernix caseosa seen under low magnification. To the naked eye, vernix appears to be a sticky white lipid paste. (b) The same vernix viewed by phase contrast microscopy. A dense packing of fetal corneocytes can be seen surrounded by a presumptive lipid matrix. Of particular note is the observation that vernix caseosa is primarily a cellular moiety. Journal of Investigative Dermatology 2000 115, 875-881DOI: (10.1046/j.1523-1747.2000.00134.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Transmission electron microscopy of vernix caseosa. These photomicrographs illustrate ultrastructural characteristics of both the cellular and the intercellular lipid components of vernix. Panel (a) reveals features of the numerous fetal corneocytes (*) that comprise vernix. These corneocytes are devoid of nuclei and other organelles and contain a sparse network of keratin filaments with little evidence of tonofilament orientation. In some locations, the corneocytes appear malleable as shown by bifurcation and curvature of the cornified cell envelope (arrowheads). No intercorneocyte desmosomal attachments are observed. Panels (b) and (c) are higher magnification photomicrographs that depict aspects of the intercellular lipids. Panel (b) shows amorphous intercellular lipids (#) that contain unidentified inclusion bodies presumably of a proteinaceous nature (arrows). A corneocyte (*) with its sparse tonofilament content can be seen in the bottom portion of the photomicrograph. Whereas most of the intercellular lipid is amorphous, panel (c) reveals that occasional lamellar structures (arrow) are observed between closely apposed corneocytes (*). Journal of Investigative Dermatology 2000 115, 875-881DOI: (10.1046/j.1523-1747.2000.00134.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Cryoscanning electron microscopy of vernix caseosa. The orientation of fetal corneocytes (*) embedded within the lipid matrix of vernix caseosa is depicted at (a) low and (b) high magnification. Panel (a>) Shows a stacking arrangement of the fetal corneocytes, whereas (b) illustrates that the lipid covering the corneocytes is arranged in a smooth leaflet-like orientation (arrowheads). A corneocyte (*) is depicted at the left-hand side of the photomicrograph. Because this portion of the cell is devoid of lipid covering and partially fractured, the interior tonofilaments are exposed and present an irregular surface. Panel (c) reveals granular inclusions in the intercorneocyte lipid (arrows). Presumably, these inclusions are the same type of structures that were noted in Figure 5(b). The asterisks denote lipid-covered corneocytes. Journal of Investigative Dermatology 2000 115, 875-881DOI: (10.1046/j.1523-1747.2000.00134.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 7 Cryoscanning electron microscopy and elemental analysis of vernix caseosa with emphasis on the water content and orientation. Specimens of vernix caseosa were cryofractured and imaged by cryoscanning electron microscopy. These photomicrographs illustrate the orientation of water within vernix. (a) Depicts a single corneocyte (*) that is partially fractured to reveal intermediate filaments within the cell. Using elemental analysis, the distribution of water in vernix is illustrated in (b). The image in the upper left panel of (b) depicts the normal appearance of a single corneocyte by cryoscanning electron microscopy. This corneocyte is indicated throughout the four panels in (b) by asterisks. Corresponding X-ray maps for oxygen (lower left panel) and carbon (upper right panel) show the distribution of water and lipid, respectively. The X-ray map in the lower right panel reflects the distribution of sulfur and is used as a background element to check for X-ray absorption artifacts due to surface topography effects. These X-ray maps reveal compartmentalization of water and lipid but not sulfur. The spectrum of these data, shown in (c), illustrates the high level of oxygen (water) relative to carbon (lipid) in this specimen. Journal of Investigative Dermatology 2000 115, 875-881DOI: (10.1046/j.1523-1747.2000.00134.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions