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In Vitro - Alternative Assays

In Vitro - Alternative Assays

Committed to developing alternative testing methods; JRF Global offers a customized battery of in vitro tests for the chemical and cosmetic industry for new ingredients and finished products.

JRF has fully committed itself to the principles of the 3Rs of animal testing. These alternative methods serve to ‘Replace’ certain animal tests, help to ‘Reduce’ the number of animals needed in a test, or to ‘Refine’ an animal testing procedure in order to reduce pain and suffering.

 

JRF offers these studies in accordance with the various Global Regulatory guidelines as well as with the REACH legislation requirements.

In vitro test methods use (reconstructed) tissues, whole cells or parts of cells. Recent advances in cell-based research include the development of two-dimensional and three-dimensional cell (co)-cultures which very closely mimic the tissues in the human body.

 

Phototoxicity
JRF Global’s in vitro research team helps you with rapid tailor-made in vitro phototoxicity assays for cosmetic products and ingredients.

Phototoxicity is a possible side effect of certain topical applications and other chemical products. Application or ingestion of such photoactive substances results in skin irritation when the skin is exposed to light, usually in reaction to UV light.

 

JRF offers the 3T3 Neutral Red Uptake (NRU) test, the standard in vitro phototoxicity test. This cytotoxic test uses a Neutral Red, a cationic dye, to assess any light-induced damage caused by the test item using murine fibroblasts as a matrix. To determine viability, the murine 3T3 fibroblast cells are exposed to both the test item and UVA light, and the NRU is then compared with the NRU of another set of fibroblast cells that have been exposed only to the test item.

 

Neutral Red, being mildly cationic, are taken up and trapped in lysosymes within the cell. Phototoxic products cause the lysosymes to rupture, releasing the Neutral Red, measurement of which is used to assess the test item’s phototoxicity.

 

Alternative test for Eye irritation
Eye irritation is a common side-effect of exposure to some hazardous chemical compounds. The effects of exposure to these compounds can range from temporary, mild corneal irritation to permanent blindness. Therefore, ocular irritation testing is an important testing requirement for several regulatory authorities across the world.

 

JRF currently offers BCOP test for eye irritation, while other two models are under validation:
Bovine Corneal Opacity & Permeability Test
This organotypic test utilizes bovine corneal tissue as a matrix, which is normally produced as a waste by-product in abattoirs. The tissue is then exposed to the test item, and the resulting changes in opacity and permeability of the corneal tissue is measured using the dye fluorescein.

 

In vitro Dermal Absorption
JRF Global offers tailor-made in vitro services for dermal absorption, using human skin and / or rodent skin.

 

Dermal absorption refers to the process that results in chemical substances passing through the skin barrier and enter the circulatory system. The rate of dermal absorption of the substance depends upon the length of exposure, properties of the item / formulation, dose and concentration of the item. The permeability characteristics of the skin layers stay the same, even when the skin is separated from the body. Therefore, in vitro dermal absorption study is an excellent alternative to traditional in vivo studies, without compromising on accuracy of data and measurements.

 

This study needs to be performed very precisely, requiring a combination of expert scientific skills, superior technical knowledge and attention-to-detail. Leveraging our vast scientific experience and regulatory expertise, JRF Global offers in vitro dermal absorption studies in accordance to OECD 428. Our scientists will work with you to create a customized program using human or rat skin as the matrix, whether it is a finished product or a cosmetic ingredient.

 

In vitro Skin corrosion Test
Chemical compounds may manifest a response when they interact with the skin. The response could vary in intensity, ranging from mild irritation to corrosion. A corrosive compound causes permanent skin injury in the form of visible necrosis, in both the epidermis and dermis. Skin necrosis occurs when cells die prematurely due to autolysis.

 

JRF Global offers a battery of in vitro skin corrosion tests, which can be customized as per your requirements to judge the corrosive potential of any formulation.

1. Skin corrosion using RhE
This test uses reconstructed human epidermis (RhE) as the matrix for conducting tests. RhE is developed from human-derived non-transformed epidermal keratinocytes and the final matrix is very similar to human epidermis in structure, morphology and properties. The skin corrosion potential of the test item is then measured using the MTT assay.

 

2. Skin Corrosion using Transcutaneous Electrical Resistance (TER) method
The TER method is an in vitro test that evaluates the human health endpoint corrosion. It identifies corrosive items by assessing changes caused to the normal structure and barrier function of the stratum corneum using rodent-based skin discs. The difference in permeability is measured using a dye; if the structure of the stratum corneum is compromised, the skin disc will become more permeable to the dye.

 

3. Skin corrosion using Membrane barrier test method
The Membrane Barrier test uses a synthetic macromolecular bio-molecule barrier and a chemical detection system (CDS) to analyze the corrosivity of the test item. The test identifies the level of damage caused to the membrane barrier after application of test item (similar to human skin) and measures the level of corrosion using pH-dependent color change or similar in the CDS on the other side of the barrier.

 

In vitro skin irritation - Dermal Irritation in 3D Human Tissue Constructs
Exposure of the skin to certain chemicals, could potentially cause damage to the outermost layer of the skin. When this damage is temporary and reversible, it is known as skin irritation. It is one of the important end points for regulatory authorities worldwide.

 

JRF Global offers the in vitro dermal irritation test using 3D Human Tissue Constructs. These constructs are obtained from human epithelial cells, which on differentiation form both the epidermis and a fully functional stratum corneum. The irritation potential of the chemical is ascertained by using the MTT assay, and the associated purple color change. JRF’s experienced scientific team will be happy to discuss and customize the test according to your needs.


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