Assessment of skin corrosion potential for medical devices

Reconstructed human epidermis testing according to OECD 431

At Eurofins Human Safety Testing the human skin model test can be used for detection of skin irritation potential of medical devices. The prediction and classification can be done by the assessment of the effect on EpiDermTM model or SkinEthicTM model.

The modified human skin model test for medical devices is a reliable in vitro test method and is performed in accordance with the draft ISO guideline 10993-10 Part 10 at our laboratory1.

Human skin model test: in vitro Skin corrosion (EpiDerm™ and EPISKIN-SM™)

The prediction and classification of skin corrositivity potential of substances can be performed by the assessment of the effect on EpiDerm and EPISKIN-SM™ models.

The human skin model test is validated by the EURL ECVAM (European Union Reference Laboratory for Alternatives to Animal Testing) and is performed in accordance with the OECD guidance (OECD 431) at Eurofins Human Safety Testing1 with chemicals, cosmetics or personal care products and pharmaceuticals.

Assessment of skin corrosion potential

  • Corrosion represents irreversible tissue damage of the skin (visible necrosis) after application of a substance. The underlying mechanisms are either the destruction of the skin penetration barrier or the rapid penetration of highly cytotoxic chemicals through the skin without its destruction.
  • The EpiDerm™ and EPISKIN-SM™ represent a reconstituted three-dimensional human epidermis (RhE) model which consists of human epidermal keratinocytes. This in vitro model mimics biochemical and physiological properties of the upper human skin.
  • To determine skin corrosion potential chemicals are applied directly on the skin tissue surface and the cell viability is measured by MTT assay.
  • The EpiDerm™ and EPISKIN-SM™ human skin model test can be used as a reliable in vitro method to distinguish between corrosive and non-corrosive chemicals. Additionally, sub-categorisation in "Category 1A" or a combination of sub-categories "1B" and "1C" is possible with this test².
  • The skin corrosion test can be performed subsequently to a positive skin irritation test for a final classification. If the skin corrosion test is performed at first, a negative result can be followed by an irritation test to conclude if the chemical has irritant potential or not.

Contact us to discuss your needs

skin-corrosion

Corrosion represents irreversible tissue damage of the skin (visible necrosis) after application of chemicals. The underlying mechanisms are either the destruction of the skin penetration barrier or the rapid penetration of highly cytotoxic chemicals through the skin without its destruction.

Procedure: Principles of the human skin model test

Protocol
Model

EpiDerm

EpiSkin
Supplier

MatTek

Skin Ethic
Analysis

Corrosivity potential: cytotoxicity measurement with MTT (mean tissue viability compared to negative control)
Test chemical concentrations
  • Liquids: 50 µL (undiluted)
  • Solids: 25 mg + 25 µL H2O
  • Liquids: 50 µL (undiluted)
  • Solids: 20 mg + 100 µL NaCl
Exposure time

3 min and 60 min incubation with dose groups

3 min, 60 min and 240 min incubation with dose groups
Quality controls
  • Positive control: 8 N (KOH)
  • Negative control: H2O
  • Positive control: glacial acetic acid
  • Negative control: NaCl
Pre-tests

To determine if additional controls are needed:

NSMTT*: mixing of test item with MTT medium to determine if test item alone can reduce MTT

à blue colouring: in main experiment two killed tissues treated with test item and two untreated killed tissues were added as controls

NSCliving#: mixing of test item with H2O or isopropanol to determine if strong own colour of test item can discolour at contact with this liquids

à optical discolouring (measuring of spectrum): in main experiment two living tissues without incubation with MTT medium were added as controls

NSCkilled§: if the two other controls were determined

à in main experiment two killed tissues without incubation with MTT medium were added as controls
Application

Direct topical application of chemicals on skin tissue

Two tissue replicates per dose group
Data delivery

Optical density (OD) value with microplate spectrophotometer at 570 nm

*NSMTT: non-specific MTT reduction
#NSCliving: non-specific colouring of living tissues
§NSCkilled: non-specific colouring of killed tissues

Procedure: Prediction model of the human skin model test

Prediction EpiDerm™

Prediction EPISKIN-SM™

Mean tissue viability

(% negative control)

Prediction

optional sub-categorisation

Mean tissue viability

(% negative control)

Prediction

optional sub-categorisation

Step 1

 --

< 50% after 3 min exp.

Corrosive

< 35% after 3 min exp.

Corrosive

Optional sub-category 1A (risk of over-prediction)

³ 50% after 3 min exp.

and

< 15% after 60 min exp.

Corrosive

A combination of optional sub-categories 1B and 1C

³ 35% after 3 min and < 35% after 60 min exp.

or

³ 35% after 60 min and < 35% after 4 h exp.

Corrosive

A combination of optional sub-categories 1B and 1C

³ 50% after 3 min exp.

and
³ 15% after 60 min exp.

Non-Corrosive

³ 35% after 4 h exp.

Non-Corrosive

Step 2

--

< 25% after 3 min exp.

Optional sub-category 1A

    

 

≥ 25% after 3 min exp.

 

A combination of optional sub-categories 1B and 1C

    

Exp. = exposure

Table 1: Prediction model of the corrosivity potential of the EpiDermTM and the EPISKIN-SMTM tissues dependent on the mean tissue viability.1

Based on the mean tissue viability, chemicals can be classified with the skin corrosion test into corrosive and non-corrosive and can be furthermore sub-categorised.

Eurofins data for demonstration technical proficiency of the human skin model test (EpiDerm™ and EPISKIN-SM™)

Chemical UN GHS category EF category for EpiDerm™ EF category for EPISKIN-SM™

Corrosive chemicals (sub-category 1A)

Bromoacetic acid

 1A 1A 1A

Boron trifluoride dihydrate

 1A 1A 1A

Phenol

 1A 1A 1A

Dichloroacetyl chloride

 1A 1A 1A

Corrosive chemicals (combination of sub-categories 1B and 1C)

Glyoxylic acid monohydrate

 1B and 1C 1B and 1C 1B and 1C

Lactic acid

 1B and 1C 1B and 1C 1B and 1C

Ethanolamine

 1B and 1C 1B and 1C 1B and 1C

Hydrochloric acid (14.4%)

 1B and 1C 1B and 1C 1B and 1C

Non-corrosive chemicals

Phenethyl bromide

 NC NC NC

4-Amino-1,2,4-triazole

 NC NC NC

4-(Methylthio)-benzaldehyde

 NC NC NC

Lauric acid

 NC NC NC

EF = Eurofins Munich GmbH                 NC = Non-Corrosive

Table 2: Eurofins data of in vitro skin corrosion with EpiDerm™ and EPISKIN-SM™ of twelve tested proficiency chemicals compared to the data of the OECD guideline 431 1.

In Table 2 the obtained data from the in vitro skin corrosion with EpiDerm™ and EPISKIN-SM™ of eight corrosive and four non-corrosive chemicals are shown. The prediction of all tested chemicals was correct in comparison to the classification of the OECD guideline 431.

References

  1. OECD (2016). OECD Guideline for the Testing of Chemicals. No. 431: In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method, 29 July, 2016
  2. UN (2015), United Nations Globally Harmonized System of Classification and Labelling of Chemicals (GHS), Sixth revised edition, UN New York and Geneva