Effects of staphylococcus aureus and aeroallergens on corneodesmosome expression and th2-promoting proinflammatory responses in human skin tissue ex vivo

Abstract Background: Both TGFb and estradiol (E2), a form of estrogen, are pro-fibrotic in the skin. In the connective tissue disease, systemic sclerosis (SSc), both TGFb and E2 are likely pathogenic. Yet, the regulation of TGFb in E2-induced dermal fibrosis remains ill-defined. Elucidating those regulatory mechanisms will improve the understanding of fibrotic disease pathogenesis and set the stage for developing potential therapeutics. Using E2-stimulated primary human dermal fibroblasts in vitro and human skin tissue ex vivo, we identified the important regulatory proteins for TGFb and investigated the extracellular matrix (ECM) components that are directly stimulated by E2-induced TGFb signaling.Methods: We used primary human dermal fibroblasts in vitro and human skin tissue ex vivo stimulated with E2 or vehicle (ethanol) to measure TGFb1, TGFb2 levels using quantitative PCR (qPCR). To identify the necessary cell signaling proteins in E2-induced TGFb1 and TGFb2 transcription, human dermal fibroblasts were pre-treated with an inhibitor of the extracellular signal-regulated kinase/ mitogen-activated protein kinase (ERK/MAPK) pathway, U0126. Finally, human skin tissue ex vivo was pre-treated with SB-431542, a TGFb receptor inhibitor, and ICI 182,780, an estrogen receptor α (ER α) inhibitor, to establish the effects of TGFb and ER α signaling on E2-induced collagen 22A1 (Col22A1) transcription. Results: We found that expression of TGFb1, TGFb2, and Col22A1, a TGFb-responsive gene, are induced in response to E2 stimulation. Mechanistically, Col22A1 induction was blocked by SB-431542 and ICI 182,780 despite E2 stimulation. Additionally, inhibiting E2-induced ERK/MAPK activation and early growth response 1 (EGR1) transcription prevents the E2-induced increase in TGFb1 and TGFb2 transcription and translation. Conclusions: We conclude that E2-induced dermal fibrosis occurs in part through induction of TGFb1, 2 and Col22A1, which is regulated through EGR1 and the MAPK pathway. Thus, blocking estrogen signaling and/or production may be a novel therapeutic option in pro-fibrotic diseases.

Download Full-text

The use of ex vivo human skin tissue for genotoxicity testing

Toxicology and Applied Pharmacology ◽

10.1016/j.taap.2012.03.019 ◽

2012 ◽

Vol 261 (2) ◽

pp. 154-163 ◽

Cited By ~ 22

Author(s):

Astrid A. Reus ◽

Mustafa Usta ◽

Cyrille A.M. Krul

Keyword(s):

Human Skin ◽

Ex Vivo ◽

Skin Tissue ◽

Genotoxicity Testing ◽

Human Skin Tissue

Download Full-text

Elucidating the Cellular Mechanism for E2-induced Dermal Fibrosis

10.21203/rs.3.rs-103581/v1 ◽

2020 ◽

Author(s):

DeAnna Baker Frost ◽

Alisa Savchenko ◽

Adeyemi Ogunleye ◽

Milton Armstrong ◽

Carol Feghali-Bostwick

Keyword(s):

Human Skin ◽

Ex Vivo ◽

Mapk Pathway ◽

Dermal Fibroblasts ◽

Skin Tissue ◽

Human Dermal Fibroblasts ◽

Dermal Fibrosis ◽

Tgfb Signaling ◽

Human Skin Tissue

Abstract Background: Both TGFb and estradiol (E2), a form of estrogen, are pro-fibrotic in the skin. In the connective tissue disease, systemic sclerosis (SSc), both TGFb and E2 are likely pathogenic. Yet, the regulation of TGFb in E2-induced dermal fibrosis remains ill-defined. Elucidating those regulatory mechanisms will improve the understanding of fibrotic disease pathogenesis and set the stage for developing potential therapeutics. Using E2-stimulated primary human dermal fibroblasts in vitro and human skin tissue ex vivo, we identified the important regulatory proteins for TGFb and investigated the extracellular matrix (ECM) components that are directly stimulated by E2-induced TGFb signaling.Methods: We used primary human dermal fibroblasts in vitro and human skin tissue ex vivo stimulated with E2 or vehicle (ethanol) to measure TGFb1, TGFb2 levels using quantitative PCR (qPCR). To identify the necessary cell signaling proteins in E2-induced TGFb1 and TGFb2 transcription, human dermal fibroblasts were pre-treated with an inhibitor of the extracellular signal-regulated kinase/ mitogen-activated protein kinase (ERK/MAPK) pathway, U0126. Finally, human skin tissue ex vivo was pre-treated with SB-431542, a TGFb receptor inhibitor, to establish the effects of TGFb signaling on E2-induced collagen 22A1 (Col22A1) transcription. Statistical tests used included parametric and non-parametric ANOVA, student’s T-test and Wilcoxon matched-pairs signed rank test. Significance was defined as p-value ≤ 0.05.Results: We found that TGFb1, TGFb2, and collagen 22A1 (Col22A1), a TGFb-responsive gene, are induced in response to E2 stimulation. Mechanistically, Col22A1 induction is blocked by the TGFb receptor inhibitor SB-431542, despite E2 stimulation. Additionally, inhibiting E2-induced ERK/MAPK activation and early growth response 1 (EGR1) transcription prevents E2-induced increase in TGFb1 and TGFb2 transcription and translation. Conclusions: We conclude that E2-induced dermal fibrosis occurs in part through induction of TGFb1, 2 and Col22A1, which is regulated through EGR1 and the MAPK pathway. Thus, blocking estrogen signaling and/or production may be a novel therapeutic option in pro-fibrotic diseases.

Download Full-text

Rapid Isolation of Functional ex vivo Human Skin Tissue-Resident Memory T Lymphocytes

Frontiers in Immunology ◽

10.3389/fimmu.2021.624013 ◽

2021 ◽

Vol 12 ◽

Author(s):

Weijie Du ◽

Daniel Lenz ◽

Ralf Köhler ◽

Erping Zhang ◽

Carla Cendon ◽

...

Keyword(s):

T Cells ◽

Human Skin ◽

Ex Vivo ◽

Skin Tissue ◽

Sample Collection ◽

Isolated Cells ◽

Tissue Dissociation ◽

Memory T Lymphocytes ◽

Collagenase Iv ◽

Human Skin Tissue

Studies in animal models have shown that skin tissue-resident memory T (TRM) cells provide enhanced and immediate effector function at the site of infection. However, analyses of skin TRM cells in humans have been hindered by the lack of an optimized isolation protocol. Here, we present a combinatorial strategy-the 6-h collagenase IV digestion and gentle tissue dissociation – for rapid and efficient isolation of skin TRM cells with skin tissue-specific immune features. In comparison with paired blood circulating memory T cells, these ex vivo isolated skin T cells express typical TRM cell markers and display higher polyfunctional properties. Moreover, these isolated cells can also be assessed for longer periods of time in ex vivo cultures. Thus, the optimized isolation protocol provides a valuable tool for further understanding of human skin TRM cells, especially for direct comparison with peripheral blood T cells at the same sample collection time.

Download Full-text

Elucidating the cellular mechanism for E2-induced dermal fibrosis

Arthritis Research & Therapy ◽

10.1186/s13075-021-02441-x ◽

2021 ◽

Vol 23 (1) ◽

Author(s):

DeAnna Baker Frost ◽

Alisa Savchenko ◽

Adeyemi Ogunleye ◽

Milton Armstrong ◽

Carol Feghali-Bostwick

Keyword(s):

Human Skin ◽

Ex Vivo ◽

Mapk Pathway ◽

Dermal Fibroblasts ◽

Skin Tissue ◽

Human Dermal Fibroblasts ◽

Ici 182,780 ◽

Dermal Fibrosis ◽

Human Skin Tissue

Abstract Background Both TGFβ and estradiol (E2), a form of estrogen, are pro-fibrotic in the skin. In the connective tissue disease, systemic sclerosis (SSc), both TGFβ and E2 are likely pathogenic. Yet the regulation of TGFβ in E2-induced dermal fibrosis remains ill-defined. Elucidating those regulatory mechanisms will improve the understanding of fibrotic disease pathogenesis and set the stage for developing potential therapeutics. Using E2-stimulated primary human dermal fibroblasts in vitro and human skin tissue ex vivo, we identified the important regulatory proteins for TGFβ and investigated the extracellular matrix (ECM) components that are directly stimulated by E2-induced TGFβ signaling. Methods We used primary human dermal fibroblasts in vitro and human skin tissue ex vivo stimulated with E2 or vehicle (ethanol) to measure TGFβ1 and TGFβ2 levels using quantitative PCR (qPCR). To identify the necessary cell signaling proteins in E2-induced TGFβ1 and TGFβ2 transcription, human dermal fibroblasts were pre-treated with an inhibitor of the extracellular signal-regulated kinase/mitogen-activated protein kinase (ERK/MAPK) pathway, U0126. Finally, human skin tissue ex vivo was pre-treated with SB-431542, a TGFβ receptor inhibitor, and ICI 182,780, an estrogen receptor α (ERα) inhibitor, to establish the effects of TGFβ and ERα signaling on E2-induced collagen 22A1 (Col22A1) transcription. Results We found that expression of TGFβ1, TGFβ2, and Col22A1, a TGFβ-responsive gene, is induced in response to E2 stimulation. Mechanistically, Col22A1 induction was blocked by SB-431542 and ICI 182,780 despite E2 stimulation. Additionally, inhibiting E2-induced ERK/MAPK activation and early growth response 1 (EGR1) transcription prevents the E2-induced increase in TGFβ1 and TGFβ2 transcription and translation. Conclusions We conclude that E2-induced dermal fibrosis occurs in part through induction of TGFβ1, 2, and Col22A1, which is regulated through EGR1 and the MAPK pathway. Thus, blocking estrogen signaling and/or production may be a novel therapeutic option in pro-fibrotic diseases.

Download Full-text

Two-dimensional thermo-mechanical fractional responses to biological tissue with rheological properties

International Journal of Numerical Methods for Heat &amp Fluid Flow ◽

10.1108/hff-03-2021-0201 ◽

2021 ◽

Vol ahead-of-print (ahead-of-print) ◽

Author(s):

Magdy A. Ezzat ◽

Roland W. Lewis

Keyword(s):

Rheological Properties ◽

Human Skin ◽

Fractional Order ◽

Order Parameters ◽

Bioheat Transfer ◽

Skin Tissue ◽

Two Dimensional ◽

Volume Relaxation ◽

Content Type ◽

Human Skin Tissue

Purpose The system of equations for fractional thermo-viscoelasticity is used to investigate two-dimensional bioheat transfer and heat-induced mechanical response in human skin tissue with rheological properties. Design/methodology/approach Laplace and Fourier’s transformations are used. The resulting formulation is applied to human skin tissue subjected to regional hyperthermia therapy for cancer treatment. The inversion process for Fourier and Laplace transforms is carried out using a numerical method based on Fourier series expansions. Findings Comparisons are made with the results anticipated through the coupled and generalized theories. The influences of volume materials properties and fractional order parameters for all the regarded fields are examined. The results indicate that volume relaxation parameters, as well as fractional order parameters, play a major role in all considered distributions. Originality/value Bio-thermo-mechanics includes bioheat transfer, biomechanics, burn injury and physiology. In clinical applications, knowledge of bio-thermo-mechanics in living tissues is very important. One can infer from the numerical results that, with a finite distance, the thermo-mechanical waves spread to skin tissue, removing the unrealistic predictions of the Pennes’ model.

Download Full-text