Assessment of the anti-C. acnes lipase activity of an ingredient

Background of anti-C. acnes lipase activity

• Cutibacterium acnes (C. acnes) is involved in the pathogenesis of acne by inducing inflammation and biofilm formation, along with other virulence factors.
• Lipases play an important role in pathogenesis of acne by hydrolysing sebum triglycerides and releasing irritating free fatty acids in the pilosebaceous follicles.
   

• Lipase has generated a high interest as a pharmacological target for antiacne drugs.

Objectives of the model

• To determine the lipase activity released by C. acnes using a fluorogenic substrate.

Our approach at Vibiosphen 

• Lipase activity was measured using the 4-methyl umbelliferyl oleate (MUO) assay (Sigma-Aldrich).
• Two C. acnes strains were used (high and low lipase expression)
• The strains were grown in brain heart infusion (BHI) broth, incubated anaerobically. Supernatants were centrifuged, filtrated and then mixed with MUO. The mixtures were incubated for 2h at 37°C in the dark. The levels of 4-methylumbelliferone released by the lipase were measured using a fluorometric microplate reader.
• Validation of the assay was performed by cultivating C. acnes in the presence of ketoconazole, a lipase expression inhibitor.
   

Outcomes of anti-C. acnes lipase activity

• This assay was developed and validated so that it can be used to evaluate anti-acne molecules in development.