01. Collagen synthesis
assay
02. Objectification of an anti-collagenase response
03. In vitro objectification of hydrating effect
04. Objectification of an anti-elastasic effect
05. Objectification of toxic outcome
06. Objectification of a modulation in turn-over concerning epithelial cells
07. .Objectification of an anti-radical effect
08. Objectification of a healing effect by analyzing a proliferation of fibroblasts
II. Evaluation of the effectiveness of active ingredients and radiation to promote cell cultures
GREDECO Exclusivity
Stemming from cultured mono-layered fibroblasts or keratinocytes, it is possible to test the effectiveness of diluted active agents in respect to culture medium or radiation (with respect to dose).
- Fibroblast culture; incubation of the cells exposed to the tested product.
- Collagen synthesis (by the kit Sircol Collagen Assay).
Fibroblastic
Fibroblastic mono-layered medium Immunochemistry anti-HLA DR
- Fibroblast culture ; contact established between the cells and the anti-collagenase agent by the culture intermediary; cellular lyse to collect enzymatic activity.
- Determination of the collagenase activity on casein electrophoresis gel.
Demonstration of collagenase activity
Demonstration of collagenase activity of type 2 and 9 metalloproteinase with electrophoresis gel
Beginning with fibroblast or human keratinocyte cultures
- Fibroblast or keratinocyte culture; incubation of cells with exposure to tested product in various concentration.
- Analysis of in vitro synthesis of glycosaminoglycans by the fibroblasts or the keratinocytes (dosage according to Blyscan Glucosaminoglycan Assay)
Evaluation of anti-elastasic activity in tubes
- Combination of anti-elastasic active with an elastase (Elastase Pig Pancrea, Synthetic Human Elastase leucocyte, N-Succinyl-Tri-Alanine-para Nitroanilide; or elastase of fibroblastic origin) in presence of a substrate
- Spectrophotometric lecture of elastasic activity at 410 nm.
Evaluation of anti-elastasic activity on frozen material
- Installation of frozen histological layers starting with a fragment of human skin (plastic surgery); elastase and product aimed at becoming an anti-elastasic in contact with dermal elastic fibers
- Coloration of elastic fibers with (+) catechin (photographs of histological cuts);
- Morphometric quantification of elastic fibers after analysing digital images (% of fibers in each cutaneous dermis, length and number of fibers).
Objectification of toxic outcome
Toxicity of fibroblast or keratinocyte cultures
- Fibroblast or keratinocyte cultures
- Cellular coloration by Giemsa
- MTT test of cellular viability
Analyse de la prolifération des kératinocytes
- Analysis of the proliferation of keratinocytes
- Keratinocyte culture
- Cell count after exclusion test with trypan blue
A partir de cultures de fibroblastes ou de kératinocytes. Possibilité de générer les radicaux libres selon deux systèmes :
- Utilising fibroblastic and keratinocytal cultures
- A possibility to generate free radicals according to two systems
- Generation of free radicals with the xanthine/hypoxanthine system and evaluation of cellular viability with the aid of the MTT test
- Generation of free radicals by UV radiation and evaluation of cellular viability with the aid of the MTT test
- - Fibroblastic culture
- - Cell count after exclusion test using trypan blue
Update : december 2010
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