SUMMARY OF THE CLINICAL STUDY

The effect of ENOCERIDE® on the dermis metabolism and on the epidermis renewal was investigated by in vitro assays using cultures of human fibroblasts and of human keratinocytes. The assays were performed in cultures of normal cells and in cultures of cells initially irradiated by UVA rays.
The effect of ENOCERIDE® on the dermis metabolism was evaluated according to the neosynthesis of collagens, of glycosaminoglycans and of elastin. They are the essential components of the extracellular matrix of dermis and play an important role in the mechanic and structural properties of the skin. The neosynthesis of these components was evaluated (1) by the measurement of the incorporation of tritiated proline in the newly synthesised collagens, secreted in the incubation medium of fibroblasts; (2) by the measurement of the incorporation of tritiated glucosamine in the newly synthesised glycosaminoglycans and (3) by the measurement of the incorporation of 14 carbon radiolabeled glycine in the newly synthesised elastin.

The effect of ENOCERIDE® on the epidermis renewal was evaluated according to the cell division capacities and to the clonogenic properties of human keratinocytes. The cell division was evaluated by measurement of the incorporation of tritiated thymidine in the newly synthesised DNA.The clonogenic properties of keratinocytes were evaluated according to the measurement of the number and the size of colonies obtained from keratinocytes.

ENOCERIDE® increased the neosynthesis of the secreted collagens and of glycosaminoglycans in normal human fibroblasts. These effects were not observed in the cultures of fibroblasts irradiated by W A rays. ENOCERIDE® had no effect on the neosynthesis of elastin. ENOCERIDE® increased the cell division capacities of normal human keratinocytes and of keratinocytes irradiated by UVA rays. ENOCERIDE® also increased the clonogenic properties of normal keratinocytes.

These data are in favour of a positive effect of ENOCERIDE® on the neosynthesis of the essential components of the extracellular matrix of the dermis involved in the structural properties (collagens) and in the mechanisms responsible for the skin consistency (glycosaminoglycans).ENOCERIDE® also increased the epidermis renewal. The positive effect of ENOCERIDE® on the cell division capacities was also observed in keratinocytes irradiated by WA rays, this result is in favour of a repairing effect.

General conclusion

Under our experimental conditions, ENOCERIDE® significantly increased the neosynthesis of the secreted collagens and of GAGs in normal human fibroblasts.

These effects were not observed when the experiment was performed in UVA irradiated cells. These data are in favour of a positive effect of ENOCERIDE® on the neosynthesis of the essential components of the extracellular matrix of the dermis involved in the structural properties (collagens). and in the mechanisms responsibles for the skin consistency (glycosaminoglycans).
ENOCERIDE® significantly increased the cell division of keratinocytes.

These results were both observed in normal cells and in keratinocytes irradiated by UVA rays. Turn-over of epidermis is fast, about 21 days. The cell division capacity decreased with ageing and so a decrease in the skin thickness and in the barrier function of the epidermis was observed. The positive effect of ENOCERIDE® in the cell division of keratinocytes is in favour of a beneficial effect in the control of epidermis homeostasis and in the barrier function of the skin. Because the positive effect of ENOCERIDE® in the cell division was also observed in WA-irradiated keratinocytes, a 'repairing effect' of ENOCERIDE® can be suggested. ENOCERIDE® also increased the clonogenic properties of keratinocytes. These cells, that the capacity of division decreased with ageing, are the base of the epidermis renewal.