Ex Vivo Human Skin: An Alternative Test System for Skin Irritation and Corrosion Assays

2021 ◽  
pp. 026119292110386
Author(s):  
Samara Eberlin ◽  
Gustavo Facchini ◽  
Gustavo Henrique da Silva ◽  
Samir Eberlin ◽  
Aline Rodrigues Bragatto ◽  
...  
Keyword(s):  
Human Skin ◽  
Corrosion Potential ◽  
Ex Vivo ◽  
Skin Irritation ◽  
Test System ◽  
Culture Conditions ◽  
Promising Tool ◽  
Alternative Test ◽  
Specific Culture ◽  
Skin Biology

Native human skin has been reported in the literature as being an important experimental model for studying skin biology. Studies performed by our group have shown that ex vivo skin, from elective plastic surgery, maintains the biological characteristics of native skin under specific culture conditions. As such, it might be a feasible model for the safety and efficacy testing of topical substances. While Brazil is at the forefront of global regulation implementation, Brazilian researchers are not always able to transfer certain widely used protocols to their laboratories, particularly protocols that involve the use of reconstructed tissues with limited viability, such as those for skin corrosion (OECD TG 431) and irritation testing (OECD TG 439). In this study, we investigated the applicability of the ex vivo skin model to the evaluation of irritation and corrosion potential of a number of proficiency substances described in TG 431 and TG 439. The skin fragments were standardised in size and diameter, and placed into cell culture inserts. The experimental protocol was conducted according to TG 431 and TG 439. The results obtained show that ex vivo skin could represent a promising tool for the evaluation of irritation and corrosion potential of substances (subject to inclusion and exclusion criteria), as recommended by OECD guidelines. While this is a proof-of-concept study, the use of ex vivo skin should be considered for such testing.

Ethanolamines permeate slowly across human skin ex vivo, but cause severe skin irritation at low concentrations

2019 ◽  
Vol 93 (9) ◽  
pp. 2555-2564
Author(s):  
Nancy B. Hopf ◽  
Philipp Spring ◽  
Gregory Plateel ◽  
Aurelie Berthet
Keyword(s):  
Human Skin ◽  
Ex Vivo ◽  
Skin Irritation ◽  
Low Concentrations

The Ex Vivo Skin Model as an Alternative Tool for the Efficacy and Safety Evaluation of Topical Products

2020 ◽  
Vol 48 (1) ◽  
pp. 10-22 ◽  
Author(s):  
Samara Eberlin ◽  
Michelle Sabrina da Silva ◽  
Gustavo Facchini ◽  
Gustavo Henrique da Silva ◽  
Ana Lúcia Tabarini Alves Pinheiro ◽  
...  
Keyword(s):  
Human Skin ◽  
Ex Vivo ◽  
Safety Evaluation ◽  
Test System ◽  
Dermal Matrix ◽  
Standard Size ◽  
Skin Explants ◽  
Alternative Approaches ◽  
Air Liquid Interface

The development of alternative approaches for safety and efficacy testing that avoid the use of animals is a worldwide trend, which relies on the improvement of current models and tools so that they better reproduce human biology. Human skin from elective plastic surgery is a promising experimental model to test the effects of topically applied products. As the structure of native skin is maintained, including cell population (keratinocytes, melanocytes, Langerhans cells and fibroblasts) and dermal matrix (containing collagen, elastin, glycosaminoglycans, etc.), it most closely matches the effects of substances on in vivo human skin. In this review, we present a collection of results that our group has generated over the last years, involving the use of human skin and scalp explants, demonstrating the feasibility of this model. The development of a test system with ex vivo skin explants, of standard size and thickness, and cultured at the air–liquid interface, can provide an important tool for understanding the mechanisms involved in several cutaneous disorders.

scholarly journals Safety Testing of Cosmetic Products: Overview of Established Methods and New Approach Methodologies (NAMs)

Cosmetics ◽  
2021 ◽  
Vol 8 (2) ◽  
pp. 50
Author(s):  
Manon Barthe ◽  
Clarisse Bavoux ◽  
Francis Finot ◽  
Isabelle Mouche ◽  
Corina Cuceu-Petrenci ◽  
...  
Keyword(s):  
Human Skin ◽  
Ex Vivo ◽  
Skin Irritation ◽  
Dermal Absorption ◽  
Skin Sensitization ◽  
Cosmetic Products ◽  
Animal Testing ◽  
Safety Testing ◽  
Novel Technologies ◽  
Cosmetic Ingredients

Cosmetic products need to have a proven efficacy combined with a comprehensive toxicological assessment. Before the current Cosmetic regulation N°1223/2009, the 7th Amendment to the European Cosmetics Directive has banned animal testing for cosmetic products and for cosmetic ingredients in 2004 and 2009, respectively. An increasing number of alternatives to animal testing has been developed and validated for safety and efficacy testing of cosmetic products and cosmetic ingredients. For example, 2D cell culture models derived from human skin can be used to evaluate anti-inflammatory properties, or to predict skin sensitization potential; 3D human skin equivalent models are used to evaluate skin irritation potential; and excised human skin is used as the gold standard for the evaluation of dermal absorption. The aim of this manuscript is to give an overview of the main in vitro and ex vivo alternative models used in the safety testing of cosmetic products with a focus on regulatory requirements, genotoxicity potential, skin sensitization potential, skin and eye irritation, endocrine properties, and dermal absorption. Advantages and limitations of each model in safety testing of cosmetic products are discussed and novel technologies capable of addressing these limitations are presented.

scholarly journals UV-independent induction of beta defensin 3 in neonatal human skin explants

F1000Research ◽  
2015 ◽  
Vol 3 ◽  
pp. 288 ◽  
Author(s):  
Erin Wolf Horrell ◽  
John D'Orazio
Keyword(s):  
Human Skin ◽  
Uv Radiation ◽  
Time Course ◽  
Ex Vivo ◽  
Culture Conditions ◽  
Skin Sample ◽  
Independent Manner ◽  
Standard Culture ◽  
Skin Explants ◽  
Multiple Donors

In order to determine the effect of UV radiation on β-defensin 3 (BD3) expression in human skin, freshly-isolated UV-naïve skin was obtained from newborn male infants undergoing planned circumcision.  Skin explants sustained ex vivo dermis side down on RPMI media were exposed to 0.5 kJ/m2 UVB, and biopsies were taken from the explant through 72 hours after radiation.  mRNA expression was measured by qRTPCR and normalized to TATA-binding protein.  BD3 expression at each time point was compared with an untreated control taken at time 0 within each skin sample.  Extensive variability in both the timing and magnitude of BD3 induction across individuals was noted and was not predicted by skin pigment phenotype, suggesting that BD3 induction was not influenced by epidermal melanization.  However, a mock-irradiated time course demonstrated UV-independent BD3 mRNA increases across multiple donors which was not further augmented by treatment with UV radiation, suggesting that factors other than UV damage promoted increased BD3 expression in the skin explants.  We conclude that BD3 expression is induced in a UV-independent manner in human skin explants processed and maintained in standard culture conditions, and that neonatal skin explants are an inappropriate model with which to study the effects of UV on BD3 induction in whole human skin.

scholarly journals UV-independent induction of beta defensin 3 in neonatal human skin explants

F1000Research ◽  
2014 ◽  
Vol 3 ◽  
pp. 288 ◽  
Author(s):  
Erin Wolf Horrell ◽  
John D'Orazio
Keyword(s):  
Human Skin ◽  
Uv Radiation ◽  
Time Course ◽  
Ex Vivo ◽  
Culture Conditions ◽  
Skin Sample ◽  
Independent Manner ◽  
Standard Culture ◽  
Skin Explants ◽  
Multiple Donors

In order to determine the effect of UV radiation on β-defensin 3 (BD3) expression in human skin, freshly-isolated UV-naïve skin was obtained from newborn male infants undergoing planned circumcision.  Skin explants sustained ex vivo dermis side down on RPMI media were exposed to 0.5 kJ/m2 UVB, and biopsies were taken from the explant through 72 hours after radiation.  mRNA expression was measured by qRTPCR and normalized to TATA-binding protein.  BD3 expression at each time point was compared with an untreated control taken at time 0 within each skin sample.  Extensive variability in both the timing and magnitude of BD3 induction across individuals was noted and was not predicted by skin pigment phenotype, suggesting that BD3 induction was not influenced by epidermal melanization.  However, a mock-irradiated time course demonstrated UV-independent BD3 mRNA increases across multiple donors which was not further augmented by treatment with UV radiation, suggesting that factors other than UV damage promoted increased BD3 expression in the skin explants.  We conclude that BD3 expression is induced in a UV-independent manner in human skin explants processed and maintained in standard culture conditions, and that neonatal skin explants are an inappropriate model with which to study the effects of UV on BD3 induction in whole human skin.

Neutrophil migration through in vitro interstitial matrix

1992 ◽  
Vol 50 (1) ◽  
pp. 894-895
Author(s):  
J. Roemer ◽  
S.R. Simon
Keyword(s):  
Extracellular Matrix ◽  
Smooth Muscle ◽  
Cell Migration ◽  
Smooth Muscle Cells ◽  
Inflammatory Cell ◽  
Neutrophil Migration ◽  
Culture Conditions ◽  
Aortic Smooth Muscle ◽  
Specific Culture

We are developing an in vitro interstitial extracellular matrix (ECM) system for study of inflammatory cell migration. Falcon brand Cyclopore membrane inserts of various pore sizes are used as a support substrate for production of ECM by R22 rat aortic smooth muscle cells. Under specific culture conditions these cells produce a highly insoluble matrix consisting of typical interstitial ECM components, i.e.: types I and III collagen, elastin, proteoglycans and fibronectin.

A Review of Electroporation-based Antitumor Skin Therapies and Investigation of Betulinic Acid-loaded Ointment

2018 ◽  
Vol 18 (5) ◽  
pp. 693-701
Author(s):  
Monika Bakonyi ◽  
Szilvia Berko ◽  
Gabor Eros ◽  
Gabor Varju ◽  
Cristina A. Dehelean ◽  
...  
Keyword(s):  
Betulinic Acid ◽  
High Efficiency ◽  
Antitumor Agents ◽  
Ex Vivo ◽  
Mouse Skin ◽  
Chemotherapeutic Agents ◽  
Hairless Mouse ◽  
Promising Tool ◽  
Acid Formulation ◽  
Invasive Method

Background: Electrochemotherapy is a novel treatment for cutaneous and subcutaneous tumors utilizing the combination of electroporation and chemotherapeutic agents. Since tumors have an increasing incidence nowadays as a result of environmental and genetic factors, electrochemotherapy could be a promising treatment for cancer patients. Objective: The aim of this article is to summarize the novel knowledge about the use of electroporation for antitumor treatments and to present a new application of electrochemotherapy with a well-known plant derived antitumor drug betulinic acid. For the review we have searched the databases of scientific and medical research to collect the available publications about the use of electrochemotherapy in the treatment of various types of cancer. Method: By the utilization of the available knowledge, we investigated the effect of electroporation on the penetration of a topically applied betulinic acid formulation into the skin by ex vivo Raman spectroscopy on hairless mouse skin. Results: Raman measurements have demonstrated that the penetration depth of betulinic acid can be remarkably ameliorated by the use of electroporation, so this protocol can be a possibility for the treatment of deeper localized cancer nodules. Furthermore, it proved the influence of various treatment times, since they caused different spatial distributions of the drug in the skin. Conclusion: The review demonstrates that electrochemotherapy is a promising tool to treat different kinds of tumors with high efficiency and with only a few moderate adverse effects. Moreover, it presents a non-invasive method to enhance the penetration of antitumor agents, which can offer novel prospects for antitumor therapies.

Development and Evaluation of Particulate Microcarriers of Adapalene as a Topical Delivery System

2019 ◽  
Vol 9 (3) ◽  
pp. 222-233
Author(s):  
Divya D. Jain ◽  
Namita D. Desai
Keyword(s):  
Patient Compliance ◽  
Targeted Delivery ◽  
Ex Vivo ◽  
Acne Vulgaris ◽  
Drug Loading ◽  
Topical Therapy ◽  
Skin Irritation ◽  
Topical Delivery ◽  
Diffusion Method ◽  
Topical Gel

Background: Adapalene is a promising third generation retinoid used in the topical treatment of acne vulgaris. However, the major drawback associated with conventional topical therapy of Adapalene is the ‘retinoid reaction’ which is dose-dependent and characterized by erythema, scaling and burning sensation at the application sites. Microparticulate drug delivery can play a major role in reducing side effects and providing better patient compliance due to targeted delivery. Methods: Adapalene microparticles were prepared using quasi emulsion solvent diffusion method. The effects of formulation variables including polymer ratios, amounts of emulsifier, drug loading and process variables such as stirring time and speed on the physical characteristics of microparticles were investigated. The developed microparticles were characterized by DSC and SEM. Adapalene microparticles were incorporated into Carbopol 971 NF gel for ease of topical delivery. Results: Adapalene microparticulate topical gel showed sustained drug release over 8 hours in in vitro studies. The amount of drug retained in the rat skin during ex vivo studies was higher in the microparticulate topical gel (227.43 ± 0.83 µg/cm2) as compared to the marketed formulation (81.4 ± 1.11 µg/cm2) after 8 hours indicating localized and sustained drug action that can be useful in treating acne vulgaris. The safety of optimized Adapalene gel determined by skin irritation studies performed on Sprague Dawley rats showed no irritation potential. Conclusion: Microparticles can provide promising carrier systems to deliver Adapalene, improving patient compliance due to enhanced skin deposition, localized and sustained action with reduced associated irritant effects.

Sign in / Sign up

Export Citation Format

Share Document