The human skin is essentially composed of three layers – stratum corneum, living epidermis and dermis – that fulfill a range of important functions, such as acting as a mechanical and chemical barrier against the environment and upholding homeostasis by regulating water loss. The thickness of each layer varies naturally between and within individuals due to a number of biological and environmental factors: e.g., age, body site, season, race, humidity, diurnal cycle, and health condition.
It is not straightforward to measure the skin thickness and its properties in vivo due to the diverse functions of the skin and the factors affecting the skin condition. To date, a range of different techniques – confocal Raman spectrometer [1], optical coherence tomography [2], reflectance confocal microscopy [3], ultrasound imaging [4], biopsy [5] and transepidermal water loss in combination with stratum corneum stripping [6, 7] – have been employed to determine the thickness of skin with findings that often vary from one method to another. The situation is exacerbated when attempts are made to measure the thickness of the outermost layer, the stratum corneum, because of its composition and thickness in the order of 10
In light of the diffficulties associated with measuring the thickness of the stratum corneum, we aim to (
In short, EIS measurements carried out with a SciBase II impedance spectrometer and a non-invasive probe [15] on skin are combined with an earlier derived mathematical model [8], for which closed-form analytical solutions are found in the limit of low frequencies around 1 kHz. We expect, based on the “forgiving” nature of approximations and leading-order solutions, that the solutions will be valid in a significant region around 1 kHz. The analytical solutions are then employed in conjunction with impedance measurements to determine the stratum corneum thickness and electrical material properties. Furthermore, in order to ensure the fidelity of the analytical expressions and establish their region of validity in the frequency spectra, verification with the numerical solution of the full set of equations is carried out; further validation with impedance measurements at different depth settings demonstrates the accuracy of the underlying mathematical model in the entire frequency spectra, 1 − 103 kHz, for the SciBase II impedance spectrometer and noninvasive probe.
The experimental EIS measurements are taken from a study that was carried out after ethics approval and informed consent had been obtained.
A total of 120 young volunteers – non-smokers without any known skin diseases or allergies – participated with an equal distribution of men and women at 24 ± 3 years of age; on the day of the measurements they were asked to abstain from moisturizers.
Electrical impedance measurements were performed on the volar forearm of the volunteers in 2007 with a SciBase II impedance spectrometer and a non-invasive probe [15].
The non-invasive, circular probe comprises four electrodes as illustrated in Fig. 1: two voltage injection electrodes, one current detector and one guard electrode to decrease the impact of surface leakage currents. With this design, two-point measurements can be carried out. The dimensions of the electrode and the skin layers depicted in Fig. 1 can be found in Table 1.
Schematic overview of the non-invasive electrode applied on (a-b) stratum corneum, viable skin and adipose tissue and (c) details of the mathematical Ansatz. The boundary conditions are denoted with roman numerals. Dimensions are given in Table 1.
Dimensions for the electrode and skin layers [8].
Model object | Acronym | Dimensions |
---|---|---|
Current detection | 1 mm | |
Ceramic | 0.15, 0.15, 1.9 mm | |
Guard ring | 0.3 mm | |
Secondary inject | 0.5 mm | |
Primary inject | 0.5 mm | |
Total width for domain | 20 mm | |
Electrode thickness | 0.1 mm | |
Stratum corneum thickness | 14 | |
Viable skin thickness | 1.2 mm | |
Adipose tissue thickness | 1.2 mm |
The SciBase II impedance spectrometer measures the impedance of the skin at 35 frequencies logarithmically distributed between 1.0 kHz and 2.5 MHz at five different depth settings; these are obtained by varying the applied voltage at the second injection electrode (boundary III in Fig. 1b) from 5 to 50 mV [16].
Before measurements, the skin of the subjects was inundated with a physiological saline solution of 0.9% sodium chloride mass concentration for 1 min. The relative air humidity was 36±7 %, and the ambient temperature was maintained at 21.7 ±0.3 C.
A mathematical model based on conservation of charge in both the time- and frequency-domain for an alternating, sinusoidal current in the rotation-symmetric gold-plated electrodes (EL) of the probe and the skin – stratum corneum (SC), viable skin (VS) and adipose tissue (AT) – in the vicinity of the probe was derived in our earlier work [8]. In short, the model can be expressed as follows in the frequency domain:
Here,
The material properties are given by [17]:
where
In our search for closed-form analytical solutions that could allow for estimates of the stratum corneum thickness and its electrical properties, we start by noting the well-known fact that the impedance is dominated by the stratum corneum at low frequencies around 1 kHz [18, 19], which suggests that the potential drop in the viable skin and electrodes are negligible vis-à-vis the corresponding drop in the stratum corneum around this frequency. This is indeed the case, as can be shown with the following scaling analysis for the potential drops in the various skin layers and electrodes: the potential drops can be secured from an order-of-magnitude equivalent of the total current density, Eq. 1b, in each layer as
where [
With these values, we find
Clearly, [Δ
At this stage, it is also instructive to see how the potential drops distribute themselves at the other end of the frequency spectrum of the SciBase II probe: 1 MHz. Returning to the order-of-magnitude equivalents, Eqs. 2a-2c, and employing the typical magnitudes for the electrical parameters at 1 MHz, we obtain
We have here used that [
In view of the findings at the lower limit of 1 kHz, for which the impedance of the skin is governed at leading order by the stratum corneum, we should be able to reduce the considered domain comprising three skin layers, the electrodes and the alternating current passing through them to a simple case of currents orthogonal to the electrodes passing in and out through the stratum corneum. Those currents would in turn give rise to the leading-order potential drop. From the mathematical point of view, it should thus be possible to reduce the partial differential equation to either ordinary differential or algebraic equations. We aim for the latter and proceed with the following Ansatz:
Cofficients for the material properties.
( | ( | ( | ( | ( | ( | |
---|---|---|---|---|---|---|
0 | -1.1803×101 | 2.3688×101 | 1.7402×100 | 1.7570×101 | 6.7610×101 | 7.5844×100 |
1 | 2.1404×101 | -2.7471×101 | 0 | -1.7961×101 | -7.0466×101 | 8.3142×10-2 |
2 | -9.9955×100 | 1.2952×101 | 0 | 8.5278×100 | 3.2847×101 | -7.7214×10-1 |
3 | 2.2537×100 | -3.0088×100 | 0 | -2.0255×100 | -7.7649×100 | 1.1797×10-1 |
4 | -2.5509×10-1 | 3.4167×10-1 | 0 | 2.3953×10-1 | 9.2429×10-1 | -4.0926×10-4 |
5 | 1.1516×10-2 | -1.5178×10-2 | 0 | -1.1319×10-2 | -4.4465×10-2 | -5.2423×10-4 |
Here, Δ
The area for each electrode,
In essence, Eq. 3a ensures that the total current is preserved, which can thus be seen as a representation of the conservation of charge, Eq. 1a;Eq. 3b is derived from the definition of the current density,
We have thus reduced the complex-valued partial differential equation in the rotation-symmetric domain comprising stratum corneum, viable skin and adipose tissue together with constitutive relations and boundary conditions to a system of 8 linear algebraic equations with 8 unknowns:
After some algebra, we arrive at the following closed-form expression for the electrode currents:
where
with the constant, A, given by
which is the ratio of the areas of the three inner electrodes modified with the depth of the second injection electrode (
The impedance,
where
We will later analyze the findings from the impedance 2in the form of the magnitude, 𝔐, and the phase, 𝔓, which can be expressed as
Finally, it is interesting to note that in the limit of 1 kHz the magnitude depends on the thickness of the stratum corneum, depth setting, frequency, material properties of stratum corneum, and the areas of the electrodes; whereas the phase is only a function of the frequency and material properties of the stratum corneum. Furthermore, both the magnitude and the phase are not functions of the applied voltage. This independence of the applied voltage has been confirmed by tests with the probe in the range from 50 mV to 250 mV, for which the measured impedance did not change at leading order (not shown here).
So far, we have derived closed-form expressions for the currents, impedance, magnitude and phase at frequencies around 1 kHz. Now, if we have access to an experimentally measured impedance,
here,
If either the relative permittivity and/or the conductivity is known, then we are able to estimate the thickness of the stratum corneum, after some algebra, from either the imaginary part of the measured impedance
or the real part
or as an average of the two
For verification of the analytical solution in the limit of low frequencies (
In essence, the computational domain shown in Fig. 1 was resolved with a mesh of around 105 triangular elements after a mesh-independence study to ensure mesh-independent results; the direct solver PARDISO was selected as the linear solver with a relative convergence tolerance of 10−6.
A typical run required around 2 seconds (wall-clock time) with 5 × 104 degrees of freedom (quadratic Lagrange elements) on a 2.5 GHz workstation for a given frequency.
We have thus far outlined the methods and materials that we intend to employ in order to estimate the thickness and electrical properties of the stratum corneum from EIS measurements. Before attempting those estimates, we need to verify the analytical solutions in the limit of 1 kHz and explore their region of validity; i.e., for which range of frequencies the solutions are reasonably valid. After verification, we proceed with the experimental measurements at various depth settings for further validation of the underlying mathematical formulation. Finally, we attempt to determine the material properties – resistivity and relative permittivity – as well as the stratum corneum thickness from the experimental impedance measurements.
In order to verify the analytical solutions and establish their region of validity, we compare the predicted magnitude and phase with their counterparts from the full set of equations, Eqs. 1a-1i, through the frequency range 1-103 kHz. As can be inferred from Fig. 2, the analytical solutions are able to predict the magnitude and phase of the EIS in the low to mid-frequency range up to around 102 kHz with an error less than 10%; after 102 kHz, the error increases rapidly up to around 25% and 40% for the magnitude and phase respectively.
The main causes for the increasing error with increasing frequency can be found in the underlying assumptions of the Ansatz: first, the assumption that the electrical impedance is dominated by the stratum corneum whilst the viable skin and subcutaneous fat are negligible becomes less accurate with increasing frequency, because it is well known that the impedance of the skin is governed by the stratum corneum only at lower frequencies whilst its influence decreases when the frequency increases [18, 19]; this was also demonstrated in the earlier scaling analysis. Second, the assumption that the currents pass straight through the stratum corneum is reasonable in the low- to mid-frequency range; at higher frequencies, however, the current density distributions underneath the electrodes becomes increasingly more non-uniform and thereby alter the active area of the electrodes (
Proceeding with verifying the individual currents for each electrode, as depicted in Fig. 3, we find that the real part agrees well throughout the frequency range, whereas the imaginary part starts to deviate above the earlier observed 102 kHz.
(a) The magnitude (▘) and phase (▾) from the numerical solution of the full set of equations and the analytical counterparts (lines); (b) the relative error for the analytical solution relative to the full set of equations for the magnitude (—) and phase (---).
The individual currents,
One of the reasons for the good agreement of the real part,
So far, we have verified and determined the region of validity of our Ansatz and resulting analytical closed-form expressions for the impedance measured during EIS of skin. We will now seek to further validate the mathematical model – solved numerically and analytically – by exploiting the different depth settings of the probe.
The measurements were carried out at five different depth settings,
Magnitude and phase of the of the numerical solution (...), analytical solution (−−) and the experimental data (symbols) at five depth settings
The ratio of the impedance to the numerical solution (...), analytical solution (−−) and the experimental data (symbols) at five depth settings
for the magnitude and the phase from the experimental measurements and predictions from the full model and analytical expressions. Here, several features are apparent: first and foremost is the good agreement between the predicted magnitude and phase for the depth setting,
Returning to the analytical solution for the impedance, Eq. 6, and extending it for different depth settings as
we find that the depth settings and their impact reduce to a constant by taking the ratio of impedances at different depth settings:
which agrees well with the corresponding experimentally measured impedance ratios at 1 kHz and numerical and analytical counterpart, as can be inferred from Fig. 5. The maximum relative error between the measured and predicted ratios varies between 0 and 8 %. In addition, it is likely that the experimental measurements are imprecise around 1 kHz, since there is no physical justification for the ratios to suddenly change abruptly when
A closer look at the individual currents (not shown here) reveals that the current changes direction at the second injection electrode (
From Eqs. 8a and 8b as well as by assuming the stratum corneum thickness to be 14
The median predicted (a) resistivity and (b) relative permittivity for the full set of equations (▘) with 1 standard deviation (▴), ±2 standard ±±deviations (▾) and the closed-form expressions (−) with ±1 standard deviation (−−), ±2 standard deviations (...).
The estimated thickness distributions for stratum corneum based on the real and imaginary part from EIS measurements and a combination of the two.
We recall that the resistivity captures more information of the real part of the impedance,
Finally, we estimate the thickness of the stratum corneum with Eqs. 9a-9c by assuming that the material properties are known. As illustrated in Fig. 7, the stratum corneum thickness estimate based on the real part of the impedance overestimates the skin thickness slightly, whilst the imaginary part underestimates the skin thickness somewhat; their combination, however, gives rise to an accurate distribution with 92% of the individuals between 6 and 20
At this stage, it would have been useful to have access to other types of measurements of the stratum corneum thickness of the 120 subjects to further provide evidence of the fidelity of the predictions. This is especially the case since we have taken material properties that were calibrated for a median skin thickness of 14
A mathematical model for EIS of skin has been analyzed and closed-form analytical expressions have been derived in the limit of low frequencies around 1 kHz. Their accuracy and region of validity were found to be 1 kHz to around 102 kHz from the verification of the full set of equations solved numerically as well as validation with experimentally measured impedances at different depth settings from a total of 120 subjects.
In summary, the analytical expressions were found from scaling analysis and a subsequent Ansatz that reduced the full set of equations – one complex partial differential equation with constitutive relations and boundary conditions in the stratum corneum, viable skin and adipose tissue – to a set of algebraic equations. The latter were solved, resulting in closed-form analytical expressions for the electrical impedance, magnitude, phase, electrical properties as well as the thickness of stratum corneum.
By combining experimental impedance measurements with the analytical expressions, we were able to not only estimate the electrical properties of the stratum corneum (provided the stratum corneum thickness is known) but also its thickness. The latter was carried out for all 120 subjects in the study, resulting in a stratum corneum thickness distribution with a mean of 14