המעבדה לביוטכנולוגיה ומיקרוביולוגיה של העור

המעבדה לביוטכנולוגיה ומיקרוביולוגיה של העור

The Dead Sea Skin Reaserch Institution, Ein Gedi, Israel.

Written by: Shiri Meshner, Ph.D.

The human skin is the largest organ of the human body. It functions as a physical barrier, protecting the body from the environment surrounding it. It forms the first line of defense against invading pathogens and helps regulate the body’s moisture and temperature. Additionally, the skin has a very important role as an active part of the immune system. Skin cells secrete various anti bacterial substances, and interact with immune cells to protect from foreign invaders. Efforts in skin-research have been directed towards a deeper understanding of the functions of the skin in establishing homeostasis and promoting immunity, and how these functions may be compromised in skin pathologies.

      

In the past few decades, the Dead Sea region has been established as a world-renown climatotherapy site for the treatment of various skin conditions such as psoriasis, atopic dermatitis, vitiligo, and others. Despite the enormous body of clinical data pointing to an efficacious treatment with a low incidence of side effects, not much is known about the mechanisms by which this success is achieved. In addition to its therapeutic potential, the climatic conditions of the Dead Sea area foster the growth of unique plants, bacteriae and algae adapted to live under extremely high temperatures, water scarcity, and increased salinity. These organisms may contain substances with a potentially protective or healing biological activity.

The Dead Sea Laboratory for Skin Biochemistry and Biotechnology of the Dead Sea and Arava Science Center has been established in the year 2009 with the aims to promote the understanding of the unique and remarkable biological phenomena related to the Dead Sea area. Situated in Ein Gedi, at the heart of the Dead Sea area, the laboratory employs a crew of skillful and enthusiastic researchers trained in various disciplines of biological sciences. In the past few years since its establishment, the lab has developed a specialty in skin research utilizing various methods in both cell culture and a specialized model system of 3-D human skin explants. Using these models and state of the art scientific equipment, we can quantitatively assess various biological phenomena pertaining to skin function such as cell proliferation, growth and metabolic activity, cell death, skin regeneration, immunological activity of skin, protection from environmental damage, and more. This way we can assay the activity of different agents that could potentially be used as pharmacological or cosmetic products.  Confining with new European regulations, the entire lab work is performed in vitro, avoiding the use of research animals. Despite that, the use of human skin explants model ensures that the results reflect the actual skin responses with high fidelity.

The Dead Sea Laboratory for Skin Biochemistry and Biotechnology strives to develop additional methods for the complete characterization of the different biological phenomena related to skin biology and pathology. We actively maintain and seek collaborations both with academia and with the industry. Owing to the acquisition of advanced robotic machinery we have recently upgraded our abilities to perform high throughput screening and analysis, and can now offer our experimental platform as a service for the cosmetic and pharmaceutical industry.

Additionally, the lab owns a fluorescence microscope equipped with optical sectioning and live cell imaging abilities which enable a thorough investigation into the biological phenomena we observe in our experiments as well as an assessment of the penetration of molecules and nanoparticles to human skin. We currently have ongoing collaborations with several labs of the Hebrew University in Jerusalem, AHAVA, The University of Aberdeen, Scotland, The Arab-American University of Jenin, the Palestinian Authority and the University of Science and Technology, Jordan. Our experimental platform can be tailored to the needs of our clients, and we possess means to develop additional assays that can be utilized for the study of skin biology.

 

The Dead Sea Skin Laboratory for Skin Biochemistry and Biotechnology 

Research methods summary

Cell characteristic

Parameter

Method

Measurement

Lab experience (0-5)

Tissue culture

Organ culture

Viability

Mitochondrial activity

MTT (colorimetric)

Creation of oxidized formazan in the mitochondria

5

5

Cellular reduction potential

Resazurin (colorimetric)

Reduction of resazurin to resorufin

5

5

Membrane integrity

Propidium Iodide (PI) (nuclear fluorescence)

Fluorescence increase of DNA-bound PI

5

5

Proliferation

Doubling time

Live cell imaging

Visual observation

5

0

Cell number increase

Methylene Blue (colorimetric)

Binding of Methylene Blue to DNA

2

0

DNA synthesis rate

BrdU incorporation

Immunodetection by ELISA

5

5

Ageing

Ageing and proliferation cellular markers

ß-galactosidase activity

Cleavage of                           ß-galactosidase fluorogenic substrate

2

2

P-16

Immunodetection by ELISA

2

2

P-21

Ki67

Inflammation

Pro- and anti- inflammatory cytokines

Pro- (Il-1, Il-6, TNF-alpha etc) and anti- inflammatory cytokines levels

Immunodetection by ELISA

3

5

Collagen degradation

UVB-derived collagen degradation

Measurement of active MMP-1

Western blot

1

3

Collagen synthesis

De-novo maturation of procollage

Measurement of the level of free C-Pro domain

Western blot

1

3

Measurement of the level of Procollagen mRNA

qPCR

1

1

Cell Cycle determination

FUCCI

Transfection of fluorescent cell cycle markers

Live cell fluorescent imaging microscopy

5

1

Doubling times

Live cell imaging

Visual measurement of doubling time

5

0

DNA quantification

FACS

Propidium Iodide staining

3

1

DNA synthesis

FACS

BrdU incorporation

2

1

UV(and other)-derived oxidative damage

Cellular antioxidant capacity

Oxygen radical absorbance capacity

Measurement of fluorescein oxidative degradation

2

2

Creation of H2O2

Resorufin fluorescence measurement

Measurement of oxidation of Amplex red using fluorescent plate reader

3

1

UV(and other)-induced apoptosis

Apoptosis

Caspase-3 activity

Cleavage of Ac-DEVD-AMC, a fluorogenic Caspase-3 substrate.

5

5

BCL-2

Immunodetection by ELISA

2

2

Annexin/PI live-dead-apoptotic assay

FACS

5

5

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

In Addition, we run a fully equipped molecular biology lab, including Real Time PCR that could be used for quantification of mRNA of various biological markers upon demand.

List of publications 

Portugal-Cohen M., et al. (2009) Protective effects of a cream containing Dead Sea minerals against UVB-induced stress in human skin. Experimental Dermatology

Portugal-Cohen M., et al.(2011) Skin organ culture as a model to study oxidative stress, inflammation and structural alterations associated with UVB-induced photodamage. Experimental Dermatology

Wineman E., et al. (2012) Photo-damage protective effect of two facial products, containing a unique complex of Dead Sea minerals and Himalayan actives. Journal of Cosmetic Dermatology (Accepted)