IFSCC 2021

Top 10 Best Posters Award

 

LVMH x LABSKIN’s poster on MMP-9 during skin aging awarded at the IFSCC 2021 congress

 

The poster presented by LVMH in collaboration with LabSkin at the last IFSCC [International Federation of Societies of Cosmetic Chemists] 2021 congress (which took place on-line on October 18th-28th) is one of the 10 posters selected by the jury, out of more than 200 posters presented, for their high level and scientific expertise. This award was unveiled by Frédéric Leroy, Chair of the IFSCC Scientific Committee, during the closing ceremony of the congress.  

 

 

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Study of a G-quadruplex aptamer inhibiting MMP-9 on human keratinocytes and on 3D reconstructed human skin model 

 

Kurfurst Robin^1*; Jeanneton Olivier¹ ; Isabelle Lorthois² ; Amélie Thépot² ; Morgan Dos Santos²

1 Life Sciences Department, LVMH Recherche, Saint Jean de Braye, France. 2 LabSkin Creations, Lyon, France. 

 

 

Introduction

Matrix Metalloprotease 9 (MMP-9) is strongly expressed and activated during skin remodeling or by different types of stress caused by UV, xenobiotic, smoke, pollution, oxidative stress. MMP-9 degrades various proteins of the dermal extracellular matrix (ECM) and epidermal basement membrane including type IV and V collagens, laminin and elastin leading to strong skin damage. For protecting skin and preventing skin aging, MMP inhibitors are key actives, however, for topical cosmetic use, natural, safe, specific and effective MMP inhibitors were not available. To overcome this limitation, during a previous work, aptamers targeting MMP-9 catalytic site were obtained by SELEX technology. The aim of this study was to evaluate inhibitory activity of one aptamer, on human keratinocyte culture and 3D reconstructed skin model. Histological analysis allowed us to observe the impacts of MMP-9 activation and its inhibition on ECM in the reconstructed skin.

Methods

On keratinocytes, optimal conditions for MMP-9 expression and activation by TNF-alpha and plasminogen have been determined then, inhibitory activities of MMP-9 aptamer and sc-311437 as control were studied. Human keratinocytes were seeded and treated with TNF-alpha for 2 days and then, co-treatments combining plasminogen and the aptamer or sc-311437 were performed for 48h. MMP-9 activities were studied on supernants by zymography. Gel semi-quantifications with Image J software were carried out to evaluate the inhibition induced by both compounds. LabSkin reconstructed skin model was obtained by culture of human fibroblasts during 21 days for ECM neosynthesis followed by seeding with human keratinocytes 21 days for epidermal compartment formation. On day 42, the reconstructed skin samples were systemically treated by TNF-alpha and on day 44 until day 46, treated with the combination of TNF-alpha, plasminogen and the MMP-9 aptamer or sc-311437. Reconstructed skin sections were fixed with formalin and stained with hematoxylin-phloxin-saffron for histological observations and epidermal thickness measurements. Stainings with Masson’s trichrome green light or Sirius red were carried out for ECM analysis. Subnatants were collected for zymography analysis and semi-quantifications were performed using Image J software. Results On keratinocyte culture, selected concentrations for sc-311437 was 100 nM and for the aptamer from 10 nM to 400 nM. We observed an inhibition of 60 % of MMP-9 activity with sc-311437 and 63% of inhibition at 10 nM to 80% inhibition at 400 nM with the aptamer. Based on these data, on reconstructed skin, we have arbitrarily used 200 nM for the aptamer. Zymography analysis on subnatants showed 33% inhibition for the aptamer and 37% for sc-311437. Histological analysis of skin sections indicated MMP-9 activation induced a strong decrease of ECM quantity and density in dermis and also a thinner epidermis. Strikingly, treatments with sc-311437 or aptamer allowed to significantly restore ECM quantity and density versus MMP-9 activated condition.

Discussion and Conclusion

For the first time, this original work demonstates on both keratinocytes and reconstructed skin, the positive impact of the use of aptamer to get a safe, efficient and natural MMP-9 inhibitor for protecting skin from ECM degradation and aging signs.

Keywords: MMP-9, aptamer, reconstructed skin, zymography, ECM, anti-aging 

 

Learn more about 3D Skin Aging model