Method – Skin moisturazation – OLEAMULS WS

Study design
The efficacy of the product was evaluated by comparing the results obtained after the use with the baseline data and placebo.

Sample size
Based on Bio Basic Europe experience, by considering the type of product, the objectives of the trial and taking into account any possible drop-out, the sample size is composed of 20 subjects.

Eligibility criteria
The subjects participating in the study were screened under medical supervision and enrolled according to the following inclusion criteria:
• both male and female sex;
• age between 18 and 60 years;
• good general health status/absence of psychological and/or cognitive disorders;
• absence of dermatological and allergological pathologies (cosmetological or to other specific excipients) or other pathologies (such as irritative reactions of unknown origin);
• absence of ongoing pharmacological treatments which may affect the outcome of the test;
• non-participation in other clinical trials in the previous 30 days;
• informed consent obtained.

Randomization
Not applicable.

Blindness
Not applicable.

Criteria for the subject withdrawal
The following rules were imposed on a possible subject withdrawal which may occur during the trial:
• breach of one of the inclusion/exclusion criteria;
• development of adverse effects.

Endpoints
• Quantitative endpoints

o Skin moisturisation (instrumental analysis)
Measured by CORNEOMETER CM 825 (A.U. arbitrary units)
The instrument measures the skin capacitance (quantification of the ability of the skin to collect electrical charge) by application of a high frequency electrical field.
The measurement is based on the principle that skin capacitance depends on the changes in dielectric constant, which are proportional to the water content in skin. With increasing hydration in the stratum corneum, its dielectric constant grows and therefore its capacitance; to clarify, the more the skin is hydrated, the more the stratum corneum becomes a good electric conductor.
Hence the instrument can indirectly detect skin hydration level, using arbitrary units.

o TEWL trans epidermal water loss (instrumental analysis)
Measured by TEWAMETER TM 300 (g/m² h)
The instrument measures trans epidermal water loss, or the quantity of water lost from the dermis and epidermis through the stratum corneum, in the form of water vapour. The result is expressed in grams per unit area per hour. The method gives indication about the integrity of the stratum corneum and thus on the efficacy of the barrier function of the skin. A barrier which is intact is related to low TEWL values, high TEWL values show on the other hand a higher water loss hence a low protective function of the skin.
The basal and short-term measurements are carried out in the treatment sites following a rest period of at least 20 minutes in an air-conditioned room with controlled and regulated temperature and humidity (temperature = 21°C +/- 2°C and humidity 40%-60%).

• Quantitative endpoints
The data on the quantitative endpoints were described using the normal position and dispersion measurements: mean and standard deviation/median and interquartile range.
A Shapiro-Wilk test was used to verify the normality of the endpoint variables, then a parametric two-way repeated measures analysis of variance model was applied to compare the products under study (treatment) and the effect over time.
When a significant interaction between treatment and time variables occurred, the results of the two main effects (treatment and time) were not taken into account and the considerations regarding the outcome of the analysis were drawn.
When a significant effect of the time variable and no significant interaction between treatment and time variables occurred, a Student’s t test with Bonferroni corrections was used to compare the different observation times.
When a significant effect of the treatment variable and no significant interaction between treatment and time variables occurred, a Student’s t test with Bonferroni corrections was used to compare the two treatments.
A significance level of <0.05 was considered.
Analyses were performed using RStudio 2022.02.0 © 2009-2022 RStudio, PBC.