Name: measuring and modeling contractile drying in human stratum corneum
Uploaded: 2017-03-01T10:30:00
Description: Watch this Scientific Journal Video about Measuring and Modeling Contractile Drying in Human Stratum Corneum at JoVE.com

The authors have no acknowledgements.

An exempt approval (3002-13) to carry out research using de-identified tissue samples pursuant to the Department of Health and Human Services regulations, 45 CFR 46.101(b)(4) was granted. Full thickness skin is received from elective surgery. In this article, the tissue source is 66-year-old Caucasian female breast.
1. Preparation of Elastomer Coated Coverslips
<ol>
	<li>In a 20 mL glass vial, mix 0.107 g of Sylgard 184 curing agent with 5.893 g base. The total mixture mass is 6 g with a bas...

<ol>
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In this article, we describe a technique that can be used to measure the dynamic drying behavior and mechanical properties of human SC. Previous studies have demonstrated that this technique can be used to quantify the effects of environmental conditions and chemical products commonly used in cosmetic cleansers and moisturizers on the dynamic drying behavior of SC7,8. There are a number of key steps in the protocol. Firstly, SC swells notably with water content; ...

The outer most layer of the epidermis, or stratum corneum (SC) consists of cohesive corneocyte cells surrounded by a lipid rich matrix1,2. The composition and structural integrity of SC is essential for maintaining correct barrier functionality3, which prevents invasion from microorganisms and resists both mechanical forces and excessive water loss4. The capacity of personal care products to maintain or degrade skin barrier function is of great interest to skin healthcare and the cosmetic industry5. The daily application of personal care products is known to alter the mechanical properties of the SC6,7,8. For example, surfactants contained in cosmetic cleansers can cause significant increases in the elastic modulus and a build-up of drying stresses in SC, increasing the tissue&#39;s propensity to crack7,9. Glycerol contained in nearly all cosmetic moisturizers can soften SC and decrease the build-up of drying stresses8,10,11, reducing the likelihood of tissue rupture.
The method detailed in this article is capable of quantifying the dynamic drying behavior and mechanical properties of SC drying in controlled environments7,8. Previously, this technique has been demonstrated to be capable of elucidating the effect of different cosmetic products on changes in the dynamic drying behavior and mechanical properties of SC tissue. This is achieved by quantifying drying-induced shrinkage of human SC tissue adhered to a soft elastomer substrate, fitting drying displacements with a simple contractility model, and then extracting the elastic modulus and drying stress from the fitted profile. When testing of multiple SC samples is required, this method offers a more rapid alternative to uniaxial tensometry, utilizes significantly less tissue and provides more physiologically relevant drying by preventing evaporation from the sample underside.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Silicone elastomer base</td>
			<td>Dow-Corning</td>
			<td>1064291</td>
		</tr>
		<tr>
			<td>Silicone elastomer Curing Agent</td>
			<td>Dow-Corning</td>
			<td>1015311</td>
		</tr>
		<tr>
			<td>FluoSpheres Carboxylate 0.1 &#181;m yellow green fluorescent 505/515&#160;</td>
			<td>Thermo Fisher</td>
			<td>F8803</td>
		</tr>
		<tr>
			<td>FluoSpheres Carboxylate 1 &#181;m yellow green fluorescent 505/515&#160;</td>
			<td>Thermo Fisher</td>
			<td>F8823</td>
		</tr>
		<tr>
			<td>FluoSpheres Carboxylate 1 &#181;m nile red fluorescent 535/575&#160;</td>
			<td>Thermo Fisher</td>
			<td>F8819</td>
		</tr>
		<tr>
			<td>Trypsin from porcine pancreas</td>
			<td>Sigma-Aldrich</td>
			<td>T6567</td>
		</tr>
		<tr>
			<td>Trypsin inhibitor type II-s</td>
			<td>Sigma-Aldrich</td>
			<td>T9128</td>
		</tr>
		<tr>
			<td>(3-aminopropyl)triethoxysilane</td>
			<td>Sigma-Aldrich</td>
			<td>440140</td>
		</tr>
		<tr>
			<td>Sodium tetraborate</td>
			<td>Sigma-Aldrich</td>
			<td>221732</td>
		</tr>
		<tr>
			<td>Boric acid</td>
			<td>Sigma-Aldrich</td>
			<td>B0294</td>
		</tr>
		<tr>
			<td>Phosphate buffered saline</td>
			<td>Sigma-Aldrich</td>
			<td>P7059</td>
		</tr>
		<tr>
			<td>N-(3-Dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>E7750</td>
		</tr>
		<tr>
			<td>Vortexer mixer</td>
			<td>VWR</td>
			<td>58816-123</td>
		</tr>
		<tr>
			<td>6mm diameter hole punch</td>
			<td>Sigma-Aldrich</td>
			<td>Z708860</td>
		</tr>
		<tr>
			<td>SOLA 6-LCR-SB&#160;</td>
			<td>Lummencor light engine</td>
			<td>No.3526</td>
		</tr>
		<tr>
			<td>Cfi Plan Achro Uw 1x Objective</td>
			<td>Nikon Plan UW</td>
			<td>MRL00012</td>
		</tr>
		<tr>
			<td>CFI Plan Fluor 40x Oil Objective 1.3 na - 0.20mm wd</td>
			<td>Nikon Plan Fluor</td>
			<td>MRH01401</td>
		</tr>
		<tr>
			<td>Nikon Eclipse Ti-U inverted microscope&#160;</td>
			<td>Nikon</td>
			<td>MEA53200</td>
		</tr>
		<tr>
			<td>Clara-E Camera</td>
			<td>Andor</td>
			<td>DR-328G-C02-SIL</td>
		</tr>
		<tr>
			<td>Remote Focus Attachment E-RFA Ergo Design</td>
			<td>Nikon</td>
			<td>99888</td>
		</tr>
		<tr>
			<td>Ti-S-E Motorized Stage</td>
			<td>Nikon</td>
			<td>MEC56110</td>
		</tr>
	</tbody>
</table>

measuring and modeling contractile drying in human stratum corneum

Stratum corneum (SC) is the most superficial skin layer. Its contact with the external environment means that this tissue layer is subjected to both cleansing agents and daily variations in ambient moisture; both of which can alter the water content of the tissue. Reductions in water content from severe barrier dysfunction or low humidity environments can alter SC stiffness and cause a build-up of drying stresses. In extreme conditions, these factors can cause mechanical rupture of the tissue. We have established a high throughput method of quantifying dynamic changes in the mechanical properties of SC upon drying. This technique can be employed to quantify changes in the drying behavior and mechanical properties of SC with cosmetic cleanser and moisturizer treatments. This is achieved by measuring dynamic variations in spatially resolved in-plane drying displacements of circular tissue samples adhered to an elastomer substrate. In-plane radial displacements acquired during drying are azimuthally averaged and fitted with a profile based on a linear elastic contractility model. Dynamic changes in drying stress and SC elastic modulus can then be extracted from the fitted model profiles.

Figure 1(a) shows a representative fluorescent image of an SC sample coated with fluorescent beads (section 3). The corresponding transmitted light image of the sample is shown in Figure 1(b) overlaid with a quiver plot of spatially resolved drying displacements that form after 16 h drying at 25% R.H. Due to the circular symmetry of the samples, these displacements can be azimuthally averaged. Figure 1(c) shows radial (ur, soli...

Watch this Scientific Journal Video about Measuring and Modeling Contractile Drying in Human Stratum Corneum at JoVE.com

Measuring and Modeling Contractile Drying in Human Stratum Corneum

This article describes a method of quantifying the dynamic drying behavior and mechanical properties of stratum corneum by measuring spatially resolved in-plane drying displacements of circular tissue samples adhered to an elastomer substrate. This technique can be used to measure how different chemical treatments alter drying and tissue mechanical properties.

Stratum corneum (SC) is the most superficial skin layer. Its contact with the external environment means that this tissue layer is subjected to both ...

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