Name: precision implementation of minimal erythema dose (med) testing to assess individual variation in human inflammatory response
Uploaded: 2019-10-03T13:30:00
Description: Watch this Scientific Journal Video about Precision Implementation of Minimal Erythema Dose MED Testing to Assess Individual Variation in Human Inflammatory Response at JoVE.com

This work was supported by a grant from the Virginia Tech College of Science Discovery Fund.

All methods described below including the use of human volunteers have been reviewed and approved by the local Institutional Review Board (IRB), and are in accordance with the Declaration of Helsinki and Belmont Report. All participants (N=72) signed informed consent as proscribed by the IRB protocol. Inclusion/exclusion criteria and discontinuation procedures were designed to maximize participant safety, and any deviation from these procedures should be considered in light of their impact on risk and tolerability to hum...

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Precision implementation of MED testing as described here could offer several advantages over other extant lab-based inflammatory challenges that have achieved popular use. For example, suction blister protocols17,18,19 raise a fluid-filled blister on the skin that is subsequently aspirated with a syringe to gain direct access to the cytokine microenvironment. Although skin blistering is a well-known tool for studying skin immun...

Minimal erythema dose (MED) testing is an FDA-approved procedure to evaluate cutaneous sensitivity to radiation typically in the UVB range, although the MED can be determined at other wavelengths in the UV and visible spectrum1. Erythema is defined as superficial reddening on the surface of the skin caused by engorgement of capillaries (later stages of erythema are more commonly known as sunburn). MED testing has been used extensively in the dermatology literature and clinical phototherapy settings to identify the minimal amount of ultraviolet (UV) radiation that will produce the smallest unit of measurable change in the redness of the skin. MED testing can be accomplished with a commercially available UV lamp, equivalent to what is used in most commercial tanning facilities.
MED testing involves continuous dispersal of UV radiation or light from the visible spectrum onto the surface of the skin for a predetermined length of time, with dosage schedules depending primarily on pigmentation of the skin and the intensity and type of radiation. This procedure is commonly used in clinical settings to determine dosage schedules for patients receiving UV radiation therapy for skin conditions such as psoriasis and eczema2,3. Basic procedures for determining the MED in clinical settings have been described elsewhere4, and can be used to adjust the total dosage of UV radiation upward or downward, depending on individual variation in skin sensitivity.
Skin pigmentation is perhaps the most important subject-specific variable in conducting and measuring results from the MED procedure6. This is because the duration of UV exposure required to evoke the minimal erythema response is principally determined by the lightness or darkness of the participant&#8217;s skin, as defined by the participant&#8217;s Fitzpatrick skin type (FST). FST7 is a numerical scheme for classifying human skin color. The Fitzpatrick scale is a recognized tool for dermatological research into human skin pigmentation8,9, and classifies human skin into one of six categories from lightest (FST I) to darkest (FST VI).
Darker FST typologies require longer UV duration, therefore accurate classification of FST is important. There is an extensive literature on methods for accurate assessment of FST, using a wide variety of approaches including self-report, dermatologist interview and instrumentation-based assessment. Observer ratings of FST have been shown to be correlated with current, but not natural skin color10, however FST can be determined subjectively11 using self-report via questionnaire12 and/or objective assessment via spectrophotometry. Fitzpatrick typing by spectrophotometry has been shown to correlate closely to participant self-report in a number of studies10,13,14,15.
Despite the utility and widespread use of MED testing in clinical services, this procedure has not been widely adopted in laboratory settings for measurement of individual variation in response to pro-inflammatory stimulation. The purpose of the methodology outlined here is to provide techniques and step-by-step procedures that increase the precision and reproducibility of the MED testing procedure, in order to facilitate future work in laboratory settings focused on fine-grained quantification of intra-individual variability in inflammatory response. We further provide representative results that illustrate the capability of this standardized protocol to accurately capture person-to-person variation in inflammation.

<table>
	<tbody>
		<tr>
			<td>6-aperture dose testing patch (&#8220;Cuff&#8221;)</td>
			<td>Daavlin</td>
			<td>&#160;</td>
			<td>&#160;</td>
		</tr>
		<tr>
			<td>Medical grade adhesive solvent</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Non-reflective UV proof cloth</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Radiometer</td>
			<td>SolarLight</td>
			<td>Model 6.2 UVB Meter</td>
			<td></td>
		</tr>
		<tr>
			<td>Single use aloe or burn gel</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrophotometer</td>
			<td>Konika-Minolta</td>
			<td>CM-2600D</td>
			<td></td>
		</tr>
		<tr>
			<td>Stopwatch</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>UV lamp &#8211; Fiji Sun</td>
			<td>Sperti</td>
			<td></td>
			<td>Emission spectrum 280nm-400nm, approximately 25% UVB</td>
		</tr>
		<tr>
			<td>UV-proof safety glasses (2 pair)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>UV-proof sleeve</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>White cotton gloves (2 pair)</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

precision implementation of minimal erythema dose (med) testing to assess individual variation in human inflammatory response

Minimal erythema dose (MED) testing is frequently used in clinical settings for determining the smallest amount of ultraviolet (UV) irradiation necessary to produce erythema (inflammatory reddening) on the surface of the skin. In this context, the MED is regarded as a key factor in determining starting doses for UV phototherapy for common skin conditions such as psoriasis and eczema. In research settings, MED testing also has potential to be a powerful tool for assessing within- and between-persons variation in inflammatory responses. However, MED testing has not been widely adopted for use in research settings, likely owing to a lack of published guidelines, which is a barrier to obtaining reproducible results from this assay. Also, protocols and equipment for establishing MED vary widely, making it difficult to compare results across laboratories. Here, we describe a precise and reproducible method to induce and measure superficial erythema using newly designed protocols and methods that can easily be adapted to other equipment and laboratory environments. The method described here includes detail on procedures that will allow extrapolation of a standardized dosage schedule to other equipment so that this protocol can be adapted to any UV radiation source.

The timing schedule presented in Table 1 is a novel dosage schedule that was calculated to capture the MED, on average, at the mid-point of the exposure event (i.e., aperture 3 or 4) for each FST. The basis for the calculated schedule is as follows.
Prior work has established that for individuals with FST 2, the median MED for radiation in the UVB range is 66.9 milliwatts (mW) per cm2, 77.429 mW/cm2 for FST 3 and 85.0 for FST 416. ...

Watch this Scientific Journal Video about Precision Implementation of Minimal Erythema Dose MED Testing to Assess Individual Variation in Human Inflammatory Response at JoVE.com

Precision Implementation of Minimal Erythema Dose MED Testing to Assess Individual Variation in Human Inflammatory Response

Minimal erythema dose (MED) testing is used to establish dosage schedules for ultraviolet radiation phototherapy. It can assess individual variation in inflammatory response but lacks methodology for achieving reproducible results. Here, we present a precision implementation of MED and demonstrate its ability to capture individual variation in inflammatory response.

Precision Implementation of Minimal Erythema Dose (MED) Testing to Assess Individual Variation in Human Inflammatory Response

Minimal erythema dose (MED) testing is frequently used in clinical settings for determining the smallest amount of ultraviolet (UV) irradiation necessary ...

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Minimal Erythema Dose (MED) Testing

Minimal Erythema Dose MED Testing

Ultraviolet radiation (UV) therapy is  sometimes used as a treatment for various common skin conditions, including  psoriasis, acne, and eczema. The ...

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