A constitutive model of human dermis skin incorporating different collagen fiber families

Skin tissue is a complex heterogeneous material abundant with fibers. Various models capturing its anisotropy, nonlinearity, viscoelasticity have been developed. However, the existence of multiple fiber families and the differences among them have been largely ignored. Furthermore, inhomogeneous deformation over the thickness is observed in the skin under shear deformation, which the traditional skin models do not predict. In this paper, we propose that two fiber families with distinct mechanical and structural properties exist in the skin within the framework of a general structure tensor-based constitutive strain energy model. Our constitutive model considers distinct properties of fiber families and the consequent inhomogeneous deformation in the skin, showing good agreement with in vivo measurements of human face skin.

Susceptibility of human and murine dermal fibroblasts to Herpes simplex Virus 1 in the absence and presence of extracellular matrix

Herpes simplex virus 1 (HSV-1) invades its human host via the skin and mucosa and initiates infection in the epithelium. While human and murine epidermis are highly susceptible to HSV-1, we recently observed rare infected cells in the human dermis and only minor infection efficiency in murine dermis upon ex vivo infection. Here, we investigated why cells in the dermis are so inefficiently infected and explored potential differences between murine and human dermal fibroblasts. In principle, primary fibroblasts are highly susceptible to HSV-1, however, we found a delayed infection onset in human compared to murine cells. Intriguingly, only a minor delayed onset of infection was evident in collagen-embedded compared to unembedded human fibroblasts although expression of the receptor nectin-1 dropped after collagen-embedding. This finding is in contrast to previous observations with murine fibroblasts where collagen-embedding delayed infection. The application of latex beads revealed limited penetration in the dermis which was more pronounced in human compared to murine dermis supporting the species-specific differences already observed for HSV-1 invasion. Our results suggest that the distinct organization of human and murine dermis contribute to the presence and accessibility of the HSV-1 receptors as well as to the variable barrier function of the extracellular matrix. These contributions, in turn, give rise to the inefficient viral access to cells in the dermis while dermal fibroblasts in culture are well infected. Importance Dermal fibroblasts are exposed to HSV-1 upon invasion in skin during in vivo infection. Thus, fibroblasts represent a widely used experimental tool to understand virus-host cell interactions and are highly susceptible in culture. The spectrum of fibroblasts’ characteristics in their in vivo environment, however, clearly differs from the observations under cell culture conditions implying putative variations in virus-cell interactions. This becomes evident when ex vivo infection studies in murine as well as human dermis revealed the rather inefficient penetration of HSV-1 in the tissue and uptake in the dermal fibroblasts. Here, we initiated studies to explore the contributions of receptor presence and accessibility to efficient infection of dermal fibroblasts. Our results strengthen the heterogeneity of murine and human dermis and imply that the interplay between dermal barrier function and receptor presence determine how well HSV-1 penetrates the dermis.

Label-free imaging of macrophage phenotypes and phagocytic activity in the human dermis in vivo using two-photon excited FLIM

Macrophages (MΦs) are important immune effector cells that promote (M1 MΦs) or inhibit (M2 MΦs) inflammation and are involved in numerous physiological and pathogenic immune responses. Their precise role and relevance, however, is not fully understood because of the lack of non-invasive quantification methods. Here, we show that two-photon excited fluorescence lifetime imaging (TPE-FLIM), a label-free non-invasive method, can visualize MΦs in human dermis in vivo. We demonstrate in vitro that human dermal MΦs exhibit specific TPE-FLIM properties that distinguish them from the main components of the extracellular matrix and other dermal cells. We visualized MΦs, their phenotypes and phagocytosis in the skin of healthy individuals in vivo using TPE-FLIM. Additionally, machine learning identified M1 and M2 MΦs with a sensitivity of 0.88±0.04 and 0.82±0.03 and a specificity of 0.89±0.03 and 0.90±0.03, respectively. In clinical research, TPE-FLIM can advance the understanding of the role of MΦs in health and disease.

CHANGES IN THE NUMBER OF p23-POSITIVE FIBROBLASTS IN HUMAN DERMIS WITH AGING

Целью работы стало исследование содержания белка p23 в фибробластах дермы человека от 20 нед беременности до 85 лет, а также определение участия p23 для возрастных изменений численности и пролиферации фибробластов в дерме человека. p23, ядерный антиген пролиферирующих клеток (PCNA), маркер фибробластов виментин выявляли в срезах кожи непрямым иммуногистохимическим методом. В дерме человека от 20 нед беременности до 85 лет наблюдали планомерное увеличение доли фибробластов, имеющих положительную окраску на p23. Возрастное изменение численности p23позитивных фибробластов в дерме достоверно коррелирует с возрастным уменьшением общей численности и доли PCNA-позитивных фибробластов в дерме человека, что свидетельствует об участии p23 в возрастных изменениях общей численности и доли пролиферирующих фибробластов в дерме человека. The aim of this work was to examine the content of p23 in fibroblasts of human dermis from 20 weeks of pregnancy until 85 years old, and defining of a role of p23 in age-dependent changes in the number and proliferation of fibroblasts in the dermis. p23, proliferating cells nuclear antigen (PCNA), fibroblasts marker vimentin were detected with indirect immunohistochemical technique. Results showed that portion of fibroblasts with positive staining for p23 in the dermis is gradually increased from 20 weeks of pregnancy until 85 years old. Age-related increase in a portion of fibroblasts with positive staining for p23 is significantly correlated with an age-related decrease in total number and percent of PCNA positive fibroblasts in human dermis. Age-related increase in the content of p23 in fibroblasts is involved in an age-dependent decrease in their total number and proliferation in the dermis.

Pathogenesis and Host Immune Response during Japanese Encephalitis Virus Infection

Japanese Encephalitis Virus (JEV) is a mosquito borne flavivirus infection. Transmission of JEV starts with the infected mosquito bite where human dermis layer act as the primary site of infection. Once JEV makes its entry into blood, it infects monocytes wherein the viral replication peaks up without any cell death and results in production of TNF-α.One of the most characteristics pathogenesis of JEV is the breaching of blood brain barrier (BBB). JEV propagation occurs in neurons that results in neuronal cell death as well as dissemination of virus into astrocytes and microglia leading to overexpression of proinflammatory cytokines. JEV infection results in host cells mediated secretion of various types of cytokines including type-1 IFN along with TNF-α and IFN-γ. Molecule like nitrous oxide (NO) exhibits antiviral activities against JEV infection and helps in inhibiting the viral replication by blocking protein synthesis and viral RNA and also in virus infected cells clearance. In addition, the antibody can also acts an opsonizing agent in order to facilitate the phagocytosis of viral particles, which is mediated by Fc or C3 receptor. This chapter focuses on the crucial mechanism of JEV induced pathogenesis including neuropathogenesis viral clearance mechanisms and immune escape strategies.

A New Treatment for the Reconstruction of the Medial Compartment of the Ankle: The Combined Use of Biological Materials

Fractures, especially if articular and periarticular, are frequently associated to functional and clinical disabling outcomes and chronic pain. In particular, the injuries with loss of bone, ligament, and/or tendon tissue in which the full recovery of the wound area is not obtained are the worst anatomical/pathological conditions to heal. In this study, three different biological materials were used as regenerative approaches to rebuild the medial malleolus fracture of the ankle in which loss of bone, ligament, and tendon tissue occur. In particular, the morselized human bone tissue was combined with the human dermis decellularized, both augmented with homologous platelet-rich plasma. The magnetic resonance imaging study with contrast at the follow-up showed a signal compatible with vascularization of the tissue without sign of degeneration. Our new regenerative approach in which different biological materials were combined together showed a good choice of treatment for the reconstruction of not repairable outcome of a fracture.

Colored Tattoo Ink Screening Method with Optical Tissue Phantoms and Raman Spectroscopy

Due to the increasing popularity of tattoos among the general population, to ensure their safety and quality, there is a need to develop reliable and rapid methods for the analysis of the composition of tattoo inks, both in the ink itself and in already existing tattoos. This paper presents the possibility of using Raman spectroscopy to examine tattoo inks in biological materials. We have developed optical tissue phantoms mimicking the optical scattering coefficient typical for human dermis as a substitute for an in vivo study. The material employed herein allows for mimicking the tattoo-making procedure. We investigated the effect of the scattering coefficient of the matrix in which the ink is located, as well as its chemical compositions on the spectra. Raman surface line scanning has been carried out for each ink in the skin phantom to establish the spatial gradient of ink concentration distribution. This ensures the ability to detect miniature concentrations for a tattoo margin assessment. An analysis and comparison of the spectra of the inks and the tattooed inks in the phantoms are presented. We recommend the utilization of Raman spectroscopy as a screening method to enforce the tattoo ink safety legislations as well as an early medical diagnostic screening tool.

Ballistic impact of single particles into gelatin : experiments and modeling with application to transdermal pharmaceutical delivery

The high speed penetration of particles into the human dermis is of interest for targeted drug delivery by transdermal powder injection. However, performing well-controlled single impact experiments with micron scale particles on dermal tissues is difficult. Therefore, the suitability of the use of a dimensionally scaled up 'model' system utilizing steel balls impacting a gelatin to simulate the perforation of micron sized gold particles into human skin was investigated. A finite element (FE) model of a 'calibration' system consisting of a 2 μm gold sphere impacting the human dermis at 651 m/s was used to extract the combinations of possible epidermal material properties which allowed an FE predicted penetration able to match measured data from an existing study in the literature. Novel scaling laws were developed to link the 'model' and 'calibration' systems, and impact experiments were performed on gelatins of various formulations to determine the formulation that produced a penetration which, when scaled, matched that of the calibration system. The resulting material properties of the gelatin were appropriately scaled and used to choose the best combination of skin material properties. In this manner, a quasi static elastic modulus of 2.25 MPa was found for skin, in good agreement with reported values from the literature. Further experiments were performed with steel, polymethyl-methacrylate, titanium, and tungsten carbide balls impacting the gelatin, in order to determine the effects of particle size and density on penetration depth. FE simulations of both the model and calibration systems confirmed the scaling relationships and impact behavior found in these experiments. Both the FE model and the steel-gelatin experiments were able to predict the penetration trends found by other investigators in the examination of typical particles used for vaccine delivery. It can therefore be concluded that scaled up systems utilizing ballistic gelatins can be used to investigate the performance of transdermal powder injection technology.

Ballistic Impact of Single Particles into Gelatin : Experiments and Modeling with Application to Transdermal Pharmaceutical Delivery

The high speed penetration of particles into the human dermis is of interest for targeted drug delivery by transdermal powder injection. However, performing well-controlled single impact experiments with micron scale particles on dermal tissues is difficult. Therefore, the suitability of the use of a dimensionally scaled up 'model' system utilizing steel balls impacting a gelatin to simulate the perforation of micron sized gold particles into human skin was investigated. A finite element (FE) model of a 'calibration' system consisting of a 2 μm gold sphere impacting the human dermis at 651 m/s was used to extract the combinations of possible epidermal material properties which allowed an FE predicted penetration able to match measured data from an existing study in the literature. Novel scaling laws were developed to link the 'model' and 'calibration' systems, and impact experiments were performed on gelatins of various formulations to determine the formulation that produced a penetration which, when scaled, matched that of the calibration system. The resulting material properties of the gelatin were appropriately scaled and used to choose the best combination of skin material properties. In this manner, a quasi static elastic modulus of 2.25 MPa was found for skin, in good agreement with reported values from the literature. Further experiments were performed with steel, polymethyl-methacrylate, titanium, and tungsten carbide balls impacting the gelatin, in order to determine the effects of particle size and density on penetration depth. FE simulations of both the model and calibration systems confirmed the scaling relationships and impact behavior found in these experiments. Both the FE model and the steel-gelatin experiments were able to predict the penetration trends found by other investigators in the examination of typical particles used for vaccine delivery. It can therefore be concluded that scaled up systems utilizing ballistic gelatins can be used to investigate the performance of transdermal powder injection technology.