Troubled Skin, Inflammasomes, and the Skin’s Innate

James V. Gruber (Vantage Specialties)

Historically, treatments for acne have focused on a known causal microbial agent, Cutibacterium acnes (C acnes), that is known to exist in high numbers in acne lesions It has been suggested that C. acnes, which populates the human skin microbiome typically as a non-pathological entity, becomes pathological in acne outbreaks However, the fundamental question of whether C acnes causes acne outbreaks, or whether it is simply responding to a modification of the skin’s innate immune response has not been adequately answered What is known is that most FDA-approved treatments for acne address the high levels of C acnes, which seems to effectively control acne outbreaks In 2002, researchers discovered the NOD-like receptor proteins (NLRP inflammasomes). These skin cell Terracotta Warrior-like sentinel proteins are the key that starts the engine of innate inflammation It has been now shown that C. acnes can elicit an innate immune response in both sebocytes and keratinocytes with a concomitant increase in NLRP inflammasome expression, and its subsequent activation of active Caspase 1, a potent protease that causes numerous downstream inflammatory responses So, the problem with troubled skin may not rest solely with control of the skin commensal C. acnes but may lie in understanding and controlling the skin’s innate immune response driven, in part, by the presence of pathological C acnes and its interaction with the skin’s innate immune response. This talk will address the skin’s innate immune response and focus on how certain well-known acne actives, like salicylic acid and azelaic acid, may do more than just kill C. acnes.

n James (V ince) Gruber, Ph.D.

Dr James (V ince) Gruber is the Director of New Product Development at Vantage Specialties developing new ideas and technologies for the personal care, cosmetic, and therapeutic industries Prior to joining Vantage Specialties, Dr Gruber held positions at Jeen, Botaneco, Sensient, Arch/Lonza, and Amerchol among others leading technical teams and developing active and functional ingredients He is the author of 40 refereed articles including 21 referenced in PubMed and holds 13 granted U.S. patents and additional published World Patent applications He has authored numerous edited articles and book chapters and is an author and co-editor of the book Principals of Polymer Science and Technology in Cosmetics and Personal Care

Ayman Grada (Case Wester n Reserve University School of Medicine)

Acne vulgaris is the most common reason for pediatric patients and third most common reason for adult patients to seek care from a dermatologist in the U S If not treated effectively, acne can lead to permanent scars. Acne is also associated with a significant psychosocial burden that impacts quality of life However, referring providers may be reluctant to initiate patients on acne treatment or certain prescriptions. In this presentation, we will review current temporal trends in epidemiology and burden of disease in the U S stratified by age, gender, and region We will share some insights on the evolving treatment paradigm and prescribing habits among dermatologists and other clinicians treating acne vulgaris

n Ayman Grada, M.D.

Dr. Grada is a U.S.-trained der matologist and a physician-scientist with over 17 years combined clinical practice, academic research, and regulated biopharma and biotech experience He currently serves as an Adjunct Professor of Der matology at Case Wester n Reserve University School of Medicine. Dr. Grada finished his der matology training and a postdoctoral fellowship in cutaneous wound healing at Boston University School of Medicine. He has published more than 120 articles in peer-reviewed jour nals on medical der matology, including

(continued on Page 18) acne vulgaris, actinic keratosis, wound healing, and epidemiology topics Dr Grada has also been serving as a Medical Director of Der matology at AbbVie Prior to joining AbbVie, Dr Grada served as the Head of R&D and Medical Affairs at Almirall (U.S.), focusing on acne vulgaris. Dr. Grada is an active physician member of the American Academy of Der matology (AAD), European Academy of Der matology and Venereology (EADV), Society for Investigative Der matology (SID), Inter national Society of Der matology (ISD), American Acne and Rosacea Society (AARS), and the Wound Healing Society (WHS)

Sophia Bull (Lucas Meyer Cosmetics)

Although the causes of acne are multifactorial, the involvement of microbiota is undeniable, particularly regarding the role played by Cutibacterium acnes (C. acnes). Our knowledge about this bacterium has evolved to include new understanding of its mode of action, which offers a new strategic perspective in the battle against acne C acnes hyperproliferation has long been thought to contribute to the development of acne, but recent studies now indicate that excess C. acnes colonization might not be an important factor, as a very small difference in the comparative amount of these bacteria in individuals with and without acne has been reported. Also, it was recently discovered that healthy skin and acneic skin have different C acnes ribotype distribution profiles between “acneic” and “non-acneic” strains

W ith this recent update on acne science, Lucas Meyer Cosmetics challenged the existing mechanisms of action to rethink the battle against acne with the next generation of anti-acne ingredients Composed of a unique patented lysine dendrimer obtained with a green chemistry manufacturing process, our dendrimer selectively weakens the acneic C acnes strains to favor growth of non-acneic strains to gently rebalance the acnebiome of acne-prone skin, therefore recovering healthier microbiota and reducing the appearance of acne

Sophia Bull joined Lucas Meyer Cosmetics as the Regional Marketing Manager for North America in 2019. In this role she supports customers and delivers regionalized promotions and launches She is based at the IFF headquarters in New York City After several years working in the phar maceutical industry, Sophia began her career in the personal care industry as regulatory support for Croda, building on her technical background Roles in marketing and sales followed at Croda and then at Botaneco. Sophia obtained her master’s degree in Chemistry from the University of Leeds in the United Kingdom

Poster Session Agenda:

• Highlights on a 3-D In Vitro Model Supporting the Development of a Natural Active Ingredient Dedicated to Acne Skin

– David Boudier (SILAB)

• Fluorescence EEM Spectroscopic Detection of Salicylic Acid Deposition on Skin from Wash-Off Products In Vivo

– Mark A. Davies, Ph.D. (Ashland Inc )

• The Role of a Cosmetic Ingredient Bis-Ethylhexyl Hydroxydimethoxy Benzylmalonate for Mitigating Acne and Improving the Outcome of Acne Treatment

– Howard Epstein, Ph.D. (EMD Electronics – An Affiliate of Merck KGaA)

• Multi-Targeted Solution for Mitigating Acne-Affected Skin

Martin Vethamuthu, Ph.D. (Sytheon)

• Lipidated Glycines as an Alter native to Conventional Anti-Acne Ingredients

– Sabrina Behnke, Ph.D. and Anneliese Schlick (TRI-K Industries)

• Assessing the Efficacy of a New Multifunctional Ingredient for Helping Topical Acne Treatment

– Cristiane Pacheco (Chemyunion)

• Using Bio-Identical Proteins for Acne Control (Bonus: Cruelty-Free and Sustainable!)

– Hannah Sivak, Ph.D. (Skin Actives Scientific)

• Sulfidal Colloidal Sulfur: A Cold-Processable, Upcycled, and Reduced Odor Sulfur as an Effective Choice for Acne Treatment

– Cristina Abbastante (Aurorium)

• Addressing the Challenges of Oily Skin with a Cosmetogenomic Approach

– Sally Hashemi and Michelle Yip (Seder ma, Croda Inc.)

• Unlocking the Acne Microbiome

– Patrick Gonry (Gobiotics-Ingredients)

• Understanding the Role of the Gut-Skin Axis in Acne-Prone Skin

Nicole Townsend and Ryan McLendon (Nutrafol)

• Boosting Skin’s Defense Mechanism Through Phase II Detoxification

– Nancy Bezies (Clariant Actives)

David Boudier, M. Mangier, M. Laclaverie, E. Aymard, and B. Closs (SILAB)

Acne is an inflammatory skin disease of the pilosebaceous unit, involving four essential factors: hyperseborrhea combined with a modification of sebum composition, colonization by Cutibacterium (C ) acnes, hyperkeratinization, and secreted inflammation Understanding and mimicking these compromised skin types is essential for further development of therapeutic solutions.

SILAB thus decided to develop a new in vitro 3-D model mimicking acneic skin by combining two main factors involved in the physiopathology: altered sebum composition and C. acnes invasion. Nor mal human keratinocytes were first used to generate reconstructed human epider mis (RHE) that were then left untreated (control) or treated topically with a combination of both peroxidized squalene and C acnes cultures. Once validated and considered relevant to mimic acneic skin, this model was further improved by using different phylotypes of C acnes strains collected by swabbing healthy volunteers and acneic patients. While both phylotypes IB and II did not significantly alter RHE, C. acnes IA1 strains induce major acneic skin hallmarks such as hyperkeratinization, secreted inflammation, and altered barrier function Interestingly, these results were obtained independently of the origin of IA1 phylotypes (acneic vs healthy patient), thus suggesting a role of the ecosystem in controlling C acnes virulence in healthy skin 1

This in vitro 3-D model mimicking acneic skin was used to screen several active molecules leading to the identification of a potential candidate for which the mechanism of action and phar macological

(continued on Page 20) studies were analyzed in detail The resulting active ingredient (Acnesium; INCI: Maltodextrin and Punica Granatum Pericarp Extract) restores the homeostasis of acneic skin by significantly targeting the principal abnor malities of this skin pathology. The following is a summary of the results (significant results with *** = P<0 001; ** = P<0 01; * = P<0 05):

• The activity of sebaceous gland is regulated by reducing 5α-reductase activity (–37%*), limiting lipid droplets secretion (up to –90%***), and restoring lipophagy (+121%**)

• Hyperkeratinization is reduced by limiting the synthesis of cytokeratin 5 (–85%**) and cytokeratin 16 (–92%**)

• Colonization by the virulent phylotype of C. acnes was mastered by limiting C. acnes phylotype IA1 growth by at least 50%, reducing the quorum sensing activity (–26%*), and preventing biofilm for mation (–43%**

• Inflammation is neutralized by limiting keratinocyte inflammation (CCL5: –100%*, CCL20: –78%, CXCL1: –94%**, IL-1α: –90%**, IL-8: –88%**); sebocyte inflammation (IL-6: –109%**, IL-8: –126%) and inflammation mediated by Th17 lymphocytes (IL-17: –38%**).

In conclusion, the development of a relevant 3-D in vitro model with the main characteristic components of acneic skin allowed the development of a natural active ingredient with transversal in vitro and in vivo efficacy Its therapeutic efficacy was demonstrated in a study conducted by dermatologists in adult Caucasian volunteers with slight to moderate acne. The clinical study reveals that the GEA (Global Evaluation Acne) score decreases significantly, and the number of lesions is reduced The natural active ingredient thus improves the quality and appearance of acneic skin, as well as the self- perception and quality of life of subjects Maltodextrin and Punica Granatum Pericarp Extract is an effective product combining rapidity of action and efficacy in the daily care of acneic skin

Reference

1 P Rouaud-Tinguely et al , From the morphological to the transcriptomic characterization of a compromised three-dimensional in vitro model mimicking atopic der matitis, BJD, 173(4): 1006-1014 (2015).

Mr David Boudier is a graduate of the Institut de Physiologie et de Biologie Cellulaire, University of Poitiers, France He holds an engineering degree in Cellular Biology He was a project leader in the SILAB cell culture laboratory for three years and then transitioned to the manager of the scientific communication unit for twelve years Currently, he is the Scientific Support Manager in R&D at SILAB.

Mark A. Davies (Ashland Inc.)

Non-invasive spectroscopic methods can be used to monitor the state of skin and the presence of materials deposited to the skin from personal care products This poster will focus on in vivo fluorescence spectrophotometry, which can provide chemical infor mation about endogenous skin fluorophores and deposited materials The principles of each of these techniques, along with the origins of endogenous fluorescence spectroscopic signals in skin will be reviewed. Examples will then be presented illustrating the use of fluorescence excitation-emission matrices (EEM) for the detection of salicylic acid deposited on skin Data analysis methods used in these studies will also be discussed n Mark A. Davies, Ph.D.

Dr Davies received his B A in Chemistry from New York University His work in spectroscopy began with his Ph.D. thesis project on Raman Optical Activity with Prof. Max Diem at the City University of New

York. This was followed by postdoctoral work on infrared spectroscopic studies of acyl chain confor mational disorder in lipid bilayer phase transitions with Prof Richard Mendelsohn at Rutgers University work which was continued as a Research Assistant Professor in the Biochemistry Department at Georgetown University School of Medicine He has carried out in vivo measurements of the efficacy of skin care products and cosmetics at Unilever, L’Oréal, and, presently, at Ashland Inc , where he conducts a variety of infrared and Raman spectroscopic studies, chemical mapping, and other physical measurements as appropriate to business needs

Howard Epstein (EMD Electronics – An Affiliate of Merck KGaA)

Acne is the most common skin condition that people experience An estimated 80% of people ages 11 to 30 will have at least a mild for m of acne Excessive sebum, bacteria, and dead skin cells clog pores or hair follicles creating potential for acne development. Other triggers of acne include environmental air pollution and high humidity Further, excessive proliferation of Cutibacterium acnes can alter the composition of sebum inducing the release of proinflammatory mediators by sebocytes A panel of experts recently published a review paper concluding that photoprotection can improve the outcome of acne treatment, because UV light can induce post-inflammatory hyperpigmentation/erythema and initiate acne flares. It was concluded that the use of specifically for mulated sunscreens could enhance topical therapy, decrease inflammation, and reduce the incidence of post-inflammatory hyperpigmentation/erythema.

This poster shows the results of a study assessing the ability of bis-ethylhexyl hydroxydimethoxy benzylmalonate (HDBM; RonaCare AP) to improve the outcome of volunteers with oily, blemished skin

The study was carried out as a randomized, placebo-controlled, single-blind study on 44 volunteers with blemished skin over a period of 8 weeks Sebum measurement, skin inflammation/redness scoring by an expert grader, photographic documentation, and a self-assessment regarding skin improvement and aesthetics were also conducted Treatment with 2% HDBM resulted in a significant reduction of sebum excretion and showed efficacy against inflamed/red lesions as shown by photographic documentation Efficacy and galenic perfor mance of 2% HDBM were judged to be superior to the placebo emulsion Conclusions from this and other studies is that HDMB is ideal for use in cosmetic for mulations, serves as an efficient sunscreen stabilizer and potent antioxidant and can replace BHT as an antioxidant to protect for mulations

Howard Epstein is Director of Technical Services for EMD Electronics, Surface Solutions, Philadelphia, PA an affiliate of Merck KGaA, Dar mstadt, Ger many He was a scholar in residence at the University of Cincinnati Department of Dermatology and received his Ph.D. in Phar macognosy from the Union Institute and University in Cincinnati, Ohio during that time He has worked in the cosmetics industry for many years since he began his career, for mulating cosmetics for Estée Lauder, Maybelline, Max Factor, Bausch & Lomb, and Kao Brands In addition to his interest in botanicals, Howard previously served as editor of the Jour nal of the Society of Cosmetic Science and is a member of the Inter national Academy of Der matology He is on the editorial board of the der matological jour nals Clinics in Der matology and SKINmed representing the cosmetics industry to dermatologists. Howard has authored chapters in various cosmetic technology textbooks including several chapters in Harry’s Cosmeticology and holds eight patents. He received the Maison G. deNavarre Medal Award in 2020, the society’s highest honor supporting the best interests of the cosmetics and personal care industry