The measurement and modelling of the mechanical properties of human skin in vivo--II. The model

The stress-strain relationship for human skin in vivo has a characteristic non linear shape even for low loads. Considerations are given on the basis of which a structural model has been selected, in which the mechanical properties of corrugated collagen fibrils are involved. It is found that such a model can describe the experimental stress-strain relationship surprisingly well with only three free parameters. These parameters are related to basic collagen fibril properties such as stiffness, diameter and waviness. The role of elastin is likely to be negligible for the purely elastic properties of human skin in vivo.     

The in vivo electrical parameters of toad skin

Open-circuit voltage (PD) and short-circuit current (SCC) across toad skin were studied in in vivo conditions. An improved technique for fastening a lucite chamber on the abdominal region of the animal was developed. Saline bridges (230 mM NaCl in 4% agar solution) were placed subcutaneously to make the connections between the extracellular fluid and the half-cells. A clear relationship was observed between the electrical parameters and sodium transport by the skin, since PD and SCC were related to the sodium concentration of the bathing solution, and abolished by the presence of amiloride--a specific sodium transport inhibitor in epithelia. The initial control values of SCC in vivo were higher than those in vitro, which was attributed to hormonal stimulation. However, these high initial control values of SCC in vivo fell with time, reaching steady levels after a 2 hr period. Vasopressin failed to increase SCC in vivo when the external sodium concentration was 115 mM, being effective only when the sodium concentration was low (5 mM). On the other hand, in isolated preparations vasopressin significantly promoted an increase in both PD and SCC.     

The tensiometric properties of expanded guinea pig skin

Our purpose in this study was to evaluate the tensile properties of expanded skin. In five guinea pigs, 29-cc ovoid tissue expanders were placed and sequentially expanded every 4 days until maximum volume was achieved. Five control and five expanded skins were harvested. Using an Instron tensile testing apparatus, skins were evaluated for stress-strain, maximum stiffness, and tensile strength, and the results were statistically compared. Centrally located expanded specimens demonstrated significantly weaker stress-strain values: 9.51 in.lb/in3 for expanded versus 30.11 in.lb/in3 for control (p less than 0.001). Maximum stiffness was similarly reduced: 4.56 lb/mm2 for expanded vs. 12.98 lb/mm2 for control (p less than 0.001). This is a 67.4 and 64.9 percent reduction, respectively, for the stress-strain and maximum stiffness. No statistically significant difference was seen in peripherally located expanded specimens relative to the controls: stress-strain expanded, 28.7 in.lb/in3 (p greater than 0.5); maximum stiffness expanded, 12.84 lb/mm2 (p greater than 0.5). Expanded skin demonstrated an average 35 percent reduction in tensile strength. We conclude that the tensile properties of expanded skin are significantly less than unexpanded skin and are a function of the degree of expansion.     

In vivo experimental testing and model identification of human scalp skin

A comprehensive experimental/numerical procedure is formulated and validated for the in vivo characterization of the mechanical properties of human skin and the simulation of reconstructive surgery. The procedure uses in vivo experimental tests on undermined skin flaps, which can be performed during surgery, a numerical model formulated within the framework of nonlinear finite strain elasticity and a nonlinear parameter identification technique for the calibration of the model from indirect measurements. The procedure is applied to characterize the scalp skin tested in Raposio and Nordstr?m (Skin Res. Technol. 4 (1998) 94). The skin is treated as a time independent, isotropic and hyperelastic membrane and the problem is solved through a finite element discretization. The study highlights that the model parameters can be determined with good accuracy using displacement measurements of a few points in the domain.     

The biomechanical effects of deep tissue support as related to brow and facelift procedures

The adverse effects of increased tension across a healing wound are well known. However, the effect of closing a wound in layers in order to decrease tension on the epidermis has been a source of controversy. It is hypothesized that deep tissue support decreases skin tension upon wound closure. In order to clarify this issue, a two-part study was designed to address the immediate effects of deep tissue support in vitro using fresh-frozen cadavers and in vivo on patients undergoing scheduled surgery. Closing skin tension was measured at standard reference points in coronal brow lift and rhytidectomy procedures performed with and without galeal closure and superficial musculoaponeurotic system (SMAS) procedures, respectively. Deep tissue support was found to significantly (p less than 0.05) decrease skin tension at the time of skin closure at standard reference points in coronal brow lift and rhytidectomy procedures performed on fresh-frozen cadavers. Similar significant (p less than 0.05) decreases in closing skin tension also were found in vivo in patients undergoing similar surgical procedures. Stress relaxation was not found to play a significant role in contributing to this immediate decrease in closing skin tension. It would appear, therefore, that deep tissue support, in the form of galeal closure and an SMAS procedure in coronal brow lift and rhytidectomy procedures, respectively, provides increased viscoelastic support, producing immediate significant decreases in closing skin tension in these procedures. The beneficial effects on wound healing, scar formation, tension-related trophic skin changes, and possible improved long-term results are discussed.     

Conductivities of pig dermis and subcutaneous fat measured with rectangular pulse electrical current

We examined experimentally the relationship between perpendicular and tangential electrical conductivities, σ, and peak current density J, in pig skin dermis and subcutaneous fat specimens by using a four-electrode measuring system with rectangular pulse electrical current (RPEC). We also investigated the relationship of the conductivity, σ, vs. pulse rate, f. The rates were selected at 8, 32, 64, and 128 pulses per second (pps), and the pulse width was fixed at 140 μs. These values are often used in vivo to enhance cutaneous regeneration with RPEC stimulation. It was found that the conductivities may be approximated to be for the skin dermis and for the subcutaneous fat in the conditions of this experiment. These findings implies that the conductivities of pig skin dermis and subcutaneous fat are anisotropic, i.e., σ_x = σ_y ≠ σ_z. It was also found that the conductivities are independent of current density and pulse rate in the current range from 20 μA/cm² to 120 mA/cm². © 1996 Wiley-Liss, Inc.     

A combined heat clearance method for tissue blood flow measurement.

Tissue Blood Flow is measured by applying a combined procedure of two independent approaches based on heat clearance: the Pulse Decay Method and the Continuous Method. The Pulse Method allows absolute assessment of tissue BF with no need for calibration, and can be applied only if the tissue BF is steady during the period of measurement. On the other hand, the Continuous Method enables the observation of rapid changes in tissue BF, and can be applied under non steady-state conditions. Using the combined method, a continuous quantitative measurement of transient changes in tissue BF can be obtained. For this purpose, we have developed two experimental systems consisting of independent electronic units: a Pulse Unit and a Continuous Unit. A micro-computer with dedicated software controls the operation of the electronic units and calculates tissue BF on-line. In vitro measurements are performed and demonstrate the reliability of the methods. In vivo measurements in rat brain tissue are also performed and include physiological and pharmacological changes of local tissue BF. The results of the two heat clearance methods correlate well with tissue BF values measured by a third independent method, the Hydrogen Clearance Method.     

In vivo measurements of electrical conductivity of porcine organs at low frequency: New method of measurement

Calculations of the induced currents created in the human body by external electromagnetic fields would be more accurate provided that more realistic experimental values of the electrical properties of the body were available. The purpose of this work is to experimentally obtain values for the conductivity of living organs in conditions close to the real situation. Two‐electrode in vivo measurements of the bioimpedance of some porcine organs have been performed. From these measurements and taking into account geometrical considerations, the electrical conductivity for the kidney, liver, heart, and spinal cord has been obtained and were found to be higher than the values reported in the literature. Furthermore, a new experimental procedure is proposed where the conductivity is determined from the values of the electrical potential and currents that are induced by an external electromagnetic field created by a coil placed close to the organ under study. Bioelectromagnetics 33:612–619, 2012. © 2012 Wiley Periodicals, Inc.     

Noninvasive measurement of the 308 nm LED-based UVB protection factor of sunscreens

The current method for determining the sun protection factor (SPF) requires erythema formation. Noninvasive alternatives have recently been suggested by several groups. Our group previously developed a functional sensor based on diffuse reflectance measurements with one UVB LED, which was previously evaluated on pig ear skin. Here we present the results of a systematic in vivo study using 12 sunscreens on 10 volunteers (skin types [ST] I-III). The relationship of the UVB-LED reflectance of unprotected skin and melanin index was determined for each ST. The spatial variation of the reflectance signal of different positions was analyzed and seems to be mainly influenced by sample inhomogeneity except for high-protection factors (PFs) where signal levels are close to detection noise. Despite the low-signal levels, a correlation of the measured LED-based UVB PF with SPF reference values from test institutes with R² = 0.57 is obtained, suggesting a strong relationship of SPF and LED-based UVB-PF. Measured PFs tend to be lower for increasing skin pigmentation. The sensor design seems to be suitable for investigations where a fast measurement of relative changes of PFs, such as due to inhomogeneous application, bathing and sweating, is of interest.     

Two-dimensional tension tests in plant biomechanics--sweet cherry fruit skin as a model system

Splitting of fruits is a function of two-dimensional tension caused by different growth rates of tissues and turgor, especially water uptake shortly before harvest. In order to analyse the mechanical properties of spheroid plant material close to stress-strain conditions in vivo, a new hydraulic two-dimensional testing device was set up. Sweet cherry (Prunus avium L.) fruit skin was chosen as a model system. The recorded pressure-deflection curves were non-linear, with a considerable initial "lag phase" and a distinct increasing end part. Taking into account the special geometry, these curves could be modelled with a newly developed analytical approach based on linear elastic material behaviour. The results demonstrated good correlation if a modulus of elasticity ranging from 160 to 250 MPa for the cherry fruit skin was chosen. In addition, a mean strength value of 47 MPa was calculated based on the theory of thin shells and spheres. The results are compared with mechanical data found for fruits and other plant material. In order to test the theoretical approach, two- and one-dimensional tension tests were performed on packaging PE foil, revealing a mean modulus of 171 MPa in bi-axial tension, and 193 and 242 MPa in uni-axial tension, depending on the test speed. The results demonstrate that it seems to be feasible to use this method to analyse the two-dimensional stress-strain conditions of spheroid plant materials such as cherry fruit skins. It may be applied as a tool for crop testing to elucidate the mechanical basis of cracking susceptibility of fruits.     

Weight measurement of upper eyelid gold implants for lagophthalmos in facial paralysis.

The use of the gold weight is an established procedure in the treatment of lagophthalmos and usually produces successful results. The critical technical issues are the firm suture fixation to the tarsal plate and the high location of the weight on this plate. However, the estimated weight of the implant from the trials on the skin of the upper lid fails to obtain the expected eye closure outcomes after surgical implantation on the tarsal plate. One of the main reasons could be the different curvature on the skin and on the tarsal plate of the upper eyelid. In this study, the angles between the vertical line and the surface of the outer (skin) and inner (tarsal plate) part of the upper eyelid have been analyzed when the eye is opened and closed. The results show that an addition of 0.2 g to the gold weight estimated in the trial is required to achieve a similar closure of the eye by means of the gold implant on the tarsal plate.     

New extensometer to measure in vivo uniaxial mechanical properties of human skin

Biomechanical properties of skin are important for clinical decision making as well as clinical intervention. Measuring these properties in vivo is critical for estimating dimensional behaviour of skin flap or graft after harvest. However, existing methodologies and devices often suffer from lack of standardisation and unwanted peripheral force contribution due to the deformation of surrounding tissues during measurement. This naturally leads to measurement inaccuracies and lack of reproducibility. In order to improve the measurement accuracy, a new portable extensometer, which measures the non-invasive in vivo biomechanical properties of skin, has been designed and constructed. This design incorporates three pads that attach to the skin, including a C-shaped pad to shield the force sensor from peripheral forces. Such design produces data that are significantly closer to in vitro measurements. The results have been verified by finite element analysis, and experiments on rubber sheets and pig skins. This device can be used to obtain biomechanical properties of skin that will aid doctors in measuring skin elasticity and surgical planning, especially in skin flap surgery.     

Combined in vivo confocal Raman spectroscopy and confocal microscopy of human skin

In vivo confocal Raman spectroscopy is a noninvasive optical method to obtain detailed information about the molecular composition of the skin with high spatial resolution. In vivo confocal scanning laser microscopy is an imaging modality that provides optical sections of the skin without physically dissecting the tissue. A combination of both techniques in a single instrument is described. This combination allows the skin morphology to be visualized and (subsurface) structures in the skin to be targeted for Raman measurements. Novel results are presented that show detailed in vivo concentration profiles of water and of natural moisturizing factor for the stratum corneum that are directly related to the skin architecture by in vivo cross-sectional images of the skin. Targeting of skin structures is demonstrated by recording in vivo Raman spectra of sweat ducts and sebaceous glands in situ. In vivo measurements on dermal capillaries yielded high-quality Raman spectra of blood in a completely noninvasive manner. From the results of this exploratory study we conclude that the technique presented has great potential for fundamental skin research, pharmacology (percutaneous transport), clinical dermatology, and cosmetic research, as well as for noninvasive analysis of blood analytes, including glucose.     

The adaptive value of grass traits in response to grazing

Aims Grazing is associated with several plant traits that may confer resistance to herbivores. However, cross-species analyses do not allow for the differentiation between adaptive evolution and common ancestry. In this study, we evaluated the effect of grazing on 5 morphological traits in 41 native grasses growing in natural grasslands of Uruguay and investigated whether such effects are independent of phylogeny.Methods We used data of grass species from 17 paired, grazed and ungrazed plots located in different regions of natural grasslands of Uruguay. For each species, we calculated the Grazing Response Index (GRI) and estimated the culm length, blade length, blade width, blade length/width ratio and caryopsis length. Trait values were calculated as the mean of the maximum and minimum values reported in a public database. We assessed the relationship between the GRI and the morphological traits using cross-species correlations, and we re-examined the correlations using phylogenetically controlled comparative analysis.Important findings Culm length and blade length were significantly correlated with the GRI. Species with higher culms and longer blades diminished their cover under grazing. This association remained significant after statistical control of phylogenetic relatedness among species. By contrast, blade width, blade length/width ratio and caryopsis length did not show any significant relationship with the GRI. Many studies in temperate grasslands recognized that several plant traits respond to grazing but were rarely evaluated in a phylogenetic context. Our results are consistent with the idea that grazing is a selective force with a clear effect on the evolution of grass stature, selecting smaller plants with shorter blades.     

Raft培养可诱导人正常上皮细胞分化

人的正常上皮细胞在一般的体外培养情况下很难分化成类似于机体的上皮样结构。为了得到上皮结构进行相关研究,近年来,国外建立了Raft（船式）培养方法。本文在长期从事细胞培养的工作中,摸索出了适合国内一般实验室条件的Raft细胞培养操作方案,模拟体内人皮肤结构的分化层次,以纤维细胞与胶原作为混合基质（作为上皮细胞的滋养物）,同时加入一些生长因子、激素和必需的蛋白等成分。将纤维细胞、胶原和添加成分做为基层过夜培养,次日将培养好的上皮细胞种到上面,过夜培养后即为“skin equivalent”（皮肤等价物）,将“皮肤等价物”移到一种网状金属架上以使细胞在气－液状态下培养,在这种条件下,培养中的上皮细胞与体内上皮细胞自然生长的状态、环境非常类似,在体外导致上皮细胞分化成清晰可见的层次。此项技术在临床治疗烧伤以及其它皮肤损伤的疾病方面有潜在的应用价值,也为研究皮肤的正常生理功能以及癌症的发生、发展以及治疗提供了一种模型。     

In Vivo Muscle Electroporation Threshold Determination: Realistic Numerical Models and In Vivo Experiments

In vivo electroporation is used as an effective technique for delivery of therapeutic agents such as chemotherapeutic drugs or DNA into target tissue cells for different biomedical purposes. In order to successfully electroporate a target tissue, it is essential to know the local electric field distribution produced by an application of electroporation voltage pulses. In this study three-dimensional finite element models were built in order to analyze local electric field distribution and corresponding tissue conductivity changes in rat muscle electroporated either transcutaneously or directly (i.e., two-plate electrodes were placed either on the skin or directly on the skeletal muscle after removing the skin). Numerical calculations of electroporation thresholds and conductivity changes in skin and muscle were validated with in vivo measurements. Our model of muscle with skin also confirms the in vivo findings of previous studies that electroporation “breaks” the skin barrier when the applied voltage is above 50?V.     

Influence of the growth rate calculation on the relationship between growth rate and temperature

Three calculations of the growth rate (e.g. slope of a plot of the log10 of cfu ml-1 vs time, mum of the Gompertz equation and the reciprocal of time to obtain 108 cfu ml-1) were compared for Escherichia coli TG1 growing in tryptone soy broth medium at temperatures ranging from 14 to 39 degrees C. Up to now, the influence of using such different definitions on the relationship between microbial growth rate and temperature has never been investigated. In order to compare these calculation procedures, a dimensionless analysis based on the following normalized variables, mudim = mu/muopt and Tdim = [T-Tmin]/[Topt-Tmin], was used (Dantigny 1998). The influence of suboptimal temperatures on the growth rate was represented by means of a Belehràdek-type model based on a power function law: [mudim] = [Tdim]alpha. The influence of the different growth rate calculations on the model constants was assessed. Despite the great dependence of the raw growth rate values on the calculation procedure, the dimensionless analysis demonstrated that the alpha-value is independent of the growth rate definition. This result suggests that any definition for the growth rate can be utilized in studies aimed at determining the influence of temperature on microbial growth and highlights the interest of using dimensionless variables to overcome differences in the order of magnitude of the growth rate data and to avoid confusion between definitions.     

Predicted sweat rate in fluctuating thermal conditions.

The ISO 7933 Required Sweat Rate Index is inappropriate under conditions of fluctuating exposure. Time weighted average values can introduce large underestimations as, for hot conditions, the relationship between physical stress and physiological strain is not linear. Mean skin temperature and sweat rate vary as a first order system, that is, according to an exponential averaging system, in response to a step function of the primary parameters. A very simple algorithm has been derived to take into account this exponential averaging and to predict the sweat rate at any time, taking into account the past exposure of the subject. It is based on time constants of 3 min for the mean skin temperature and of 25 min (up to a sweat rate of 50 W.m-2) and 10 min for the sweat rate. The correlation coefficient is 0.780 between observed and predicted sweat rates in independent experiments used for the validation of the model. This emphasises the necessity for exponential averaging as differences between individuals are very important and the precision of the observed data in these experiments was lower than for laboratory experimental data.     

The relationship between ram sperm head morphometry and fertility depends on the procedures of acquisition and analysis used

Sperm head morphometry is a parameter in the evaluation of semen that has been associated with fertility in two ways: comparing morphometric measures between predefined groups of fertility; or analyzing morphometric data by multivariate techniques to identify cell populations. We analyzed the morphometry of ram sperm head by three procedures and checked its relationship with male fertility. A Computer-Aided Sperm Morphometric Assessment procedure (CASMA), an image analysis software (NIS-Elements) in combination with an optical microscope (MO-NIS) and this image analysis software in combination with a scanning electron microscope (SEM-NIS) were used. Eight morphometric parameters were assessed: length, width, area, perimeter, ellipticity, form factor, elongation and regularity. We observed significant differences between the morphometric data of sperm head obtained with three study procedures. The CASMA procedure shows the highest values for all parameters and the SEM-NIS procedure the lowest. The analysis of a semen sample, when only the mean of morphometric parameters is used to describe the cell population, is too limited to interpret their fertilizing capacity. It is essential to analyze the complex structure of the samples by defining subpopulations by multivariate methods. With few exceptions, the means of each morphometric parameter differ between the three subpopulations analyzed in each procedure. Only the subpopulations obtained with the MO-NIS procedure showed a significant correlation with male fertility. In short, it is necessary to establish an instrumental standard for the analysis of sperm morphometry to obtain reliable results and we believe that the MO-NIS system presents these basic requirements.     

Properties and microstructure of thermo-pressed wheat gluten films: a comparison with cast films

Wheat gluten films were prepared by thermo-pressing, and their mechanical properties were compared to those of cast films. The stress-strain relationship was established for films with various amounts of glycerol. Both relationships were quite different, revealing a different network organization. Thermo-pressed films presented higher stress values than cast films, but the effect of the glycerol amount was similar in both cases, an increase of the glycerol amount leading to a decrease of both films stress. The glycerol influence on the strain at break of thermo-pressed films was very limited, with strain values reaching a maximum around 200%. The role of disulfide bridges on themomoulded films mechanical properties was investigated, and it was shown that some rearrangements and a significative protein insolubilization occurred during the process. The effective flow porosity of the protein network for thermo-pressed films was estimated by water capillary rise measurements to about 7%. Scanning electron microscopy was used to obtain some information about the microstructure of both cast and thermo-pressed films.