11th Scientific Symposium LVMH Recherche

Skin Rejuvenation
London UK Thursday 27th October 2011

11th Scientific Symposium LVMH Recherche

Skin Rejuvenation
London UK Thursday 27th October 2011

The British Library

Dr. Barry Knight Head of Conservation Research
The British Library is the national library of the United Kingdom, a major research library holding over 150 million items from every country in the world, in many formats, print or digital: books, manuscripts, newspapers, journals, magazines, sound and music recordings, videos, maps, stamps, prints, drawings For many years, the British Library has also developed a conservation research strategy and collaborative applied research programmes to maximize the future availability of collections. The British Library is a unique and fabulous place in the worldwide cultural and information network, an institution which supports research, guarantees access for future generations to the worlds knowledge, and enriches the cultural life of the nation.

Welcome
Frdric Bont Director of Scientific Communication
Dear Colleagues, It is my great pleasure and honour to welcome you to the 2011 LVMH Recherche Symposium held at the British Library in London. In many countries, in the next 50 years, about one-third of women will be over 50. Appearance and skin vitality are and will remain a subject of primary importance in daily life and for social well being. If there is beauty to be found in every human face, the cosmeticians role is to provide skin care and makeup products that allow a womans facial features to work in concert and reflect light with more harmony and equilibrium. Scientists are looking for new strategies able to regenerate the health and youth of the skin by producing a durable effect below the skins surface. Today, selected scientific presentations will cover a wide range of topics and will present the latest scientific discoveries in anti-aging and skin rejuvenation research. Rejuvenation is the central focus of thousands research projects in laboratories in the field of genetics, cellular and molecular biology, active ingredients, reprogramming strategies and formulation. The goal of these projects is to help women to achieve naturally radiant skin and to reestablish youthful facial characteristics. Let me remind you that the aim of this LVMHRecherche symposium is to gather together leading scientists in order to discuss one of the major themes in the field of cosmetics and to generate transdisciplinary links. It is a great privilege to hold this symposium at the British Library, which occupies a unique and inspiring place in the worlds cultural and information network, an institution that supports research, guarantees access to world knowledge to future generations and enriches the cultural life of nations.

that of phytochemists in our ethnobotanical network, aim to identify biological targets and develop active Executive Vice-President R&D ingredients and formulas to help maintain the beauty LVMH Recherche and youth of the skin. Our Philosophy: By combining our knowledge with information from Research for emotion the fields of sociology, psychology and neurocosmetics, and validating our results in clinical studies and Healthy skin is our main source of inspiration. To consumer panels worldwide, we seek to reach taibetter understand it, our Research and Development lored cosmetics solutions. Center has developed a comprehensive set of investi- Deeply rooted in a culture of sharing knowledge and gative tools in immunohistochemistry, cell culture, skin respect of everyones skills, our research philosophy is reconstruction, skin biopsy, DNA chips, proteomics, enriched by our own expertise as well as that of the advanced imaging and sensory maps, among others. international experts with whom we work. Our daily goal is to better understand the biological mechanisms that govern skins appearance, evolution and rhythm. Research by our biologists, combined with

eric perrier

processes that restructure the skins composition to Scientific adviser restore its lost momentum. One could say that this is a drive towards the future that restores the conditions of a youthful past. Skin has a natural potential to conserve its beauty, Recently new terms have cropped up in the field of and probably contains the secrets of prolonged youth. cosmetics to describe new concepts, such as dermaSome cells naturally live for many years and others toporosis, while other terms have remained the same have an extraordinary power of renewal. Moreover, it but no longer have quite the same meaning. The area appears that our biological clock is part of a process of hydration, for example, which was revolutionized of aging whose activation is not inevitable, and we can by the discovery of aquaporins, for which Peter Agre slow down its pendulum. Even better, it is becoming earned a Nobel Prize in 2003, has been important possible to prevent our biological clock from cutting to the formulation of our products for some years. time short and to compel it to reverse its conse- The regulation of proteasome (whose discovery also quences. Thus a new era that goes beyond anti-aging earned a Nobel Prize in 2004) represents another is emerging, an era of rejuvenation. breakthrough in our formulations. Among antioxyLVMH Research is aimed at making advances in dants, complex molecules were isolated from specific understanding skin composition, but also in the areas vine shoots of rare french vines and some polypheof the protection, repair and the enzymes of the skin nolic trimers (miyabenol) show remarkable biological slowing or stimulating their production as well as activities to fight skin ageing. Our research laboratory is currently working on many projects whose aim is to find active ingredients that would help in skin rejuvenation. Genetic sites involved

Max santoul

in aging may be targeted, stem cells reactivated, enzymatic processes optimized. For example, of particular interest are sirtuins, cellular enzymes in close contact with proteins that manage our genetic inheritance and affect cellular metabolism by regulating the expression of certain genes. Current research has developed to the point where a youthful appearance is a right to which we can all aspire, and the protection of an organ as precious as the skin is becoming a duty. This living envelope, which has influenced the evolution of our species since the dawn of humanity, clothes our bodies in nudity.This soft, smooth, durable, expandable, waterproof garment that protects us from a hostile environment must stand the test of time. It holds an exceptional power of longevity and perhaps, secretly, a universal power of repair and renewal. If we had to summarize the history of cosmetics, we would say that it dates back to the earliest civilizations. For millennia cosmetics were devoted to beautification, then they entered the era of anti-aging during

the twentieth century, with an emphasis on repair and protection (antioxydants are well-established examples), and now they are moving towards the future, focusing on rejuvenation, with research on stem cells and agents capable of reversing the course of time. Through this symposium we hope to share with you our enthusiasm for the latest advances in this exciting field.

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Program
Welcome
Eric Perrier, Frdric Bont, LVMH Recherche, Saint Jean de Braye, France

Introduction
Dr. Barry Knight, Head of Conservation Research, British Library, London, UK

Prospects for truly comprehensive repair of aged skin

Dr. Aubrey de Grey, Chief Science Officer, SENS Foundation, Mountain View, USA

A dermatologists view on skin rejuvenation

Dr. Leslie Baumann, Chief Executive Officer, Baumann Cosmetic & Research Institute, Miami, FL, USA

Stem cells and skin rejuvenation

Pr. Carlo Pincelli, Professor of Dermatology, School of Biosciences and Biotechnologies, University of Modena and Reggio Emilia, Italy

Translational reverse-aging research: latest advances

Melanie Swan, MBA, Futurist and Applied Genomics Expert, MS Futures Group, DIYgenomics, Palo Alto, USA

Skin-cell rejuvenation
Dr. Carine Nizard, Bio-Science Innovation Manager, LVMH Recherche, Saint Jean de Braye, France

Signalling pathways and rejuvenation: risks and opportunities

Oleg Kvitko, Ph.D., Leading scientist, Institute of Genetics and Cytology, National Academy of Sciences, Minsk, Belarus

Controlled wound healing for skin rejuvenation: recent progress and future challenges
Dr. Laure Ritti, Research Investigator, Department of Dermatology Photoaging and Aging Research Program, Ann Arbor, University of Michigan, USA

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ventions to repair and/or obviate that damage. He has Chief Science Officer, SENS developed a possibly comprehensive plan for such repair, termed Strategies for Engineered Negligible Foundation, Mountain View, USA Senescence (SENS), which breaks aging down into Dr. Aubrey de Grey is a biomedical gerontologist seven major classes of damage and identifies detailed based in Cambridge, UK, and is the Chief Science approaches to addressing each one. A key aspect of Officer of SENS Foundation, a California-based cha- SENS is that it can potentially extend healthy lifespan rity dedicated to combating the aging process. He without limit, even though these repair processes will is also Editor-in-Chief of Rejuvenation Research, the probably never be perfect, as the repair only needs to worlds highest-impact peer-reviewed journal focused approach perfection rapidly enough to keep the oveon intervention in aging. He received his BA and Ph.D. rall level of damage below pathogenic levels. Dr. de from the University of Cambridge in 1985 and 2000 Grey has termed this required rate of improvement respectively. His original field was computer science, of repair therapies longevity escape velocity. Dr. de and he did research in the private sector for six years Grey is a Fellow of both the Gerontological Society in the area of software verification before switching of America and the American Aging Association, and to biogerontology in the mid-1990s. His research sits on the editorial and scientific advisory boards of interests encompass the characterisation of all the numerous journals and organisations. accumulating and eventually pathogenic molecular and cellular side-effects of metabolism (damage) that constitute mammalian aging and the design of inter-

aubrey de grey

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Prospects for truly comprehensive repair of aged skin

Though aging of the skin is not in itself a life-threatening process, the immense demand for skin care products demonstrates how important skin aging is in diminishing self-esteem and quality of life. In spite of continued progress in improving the efficacy of skin rejuvenation methods, existing techniques remain far from fully effective. Where will future major breakthroughs in addressing skin aging come from? There are two distinct classes of obstacle to achieving truly comprehensive skin rejuvenation. Firstly, the skin is a rather complex tissue, incorporating two very different layers of cells (the dermis and the epidermis), each of which possesses multiple cell types and which are separated by an extracellular lamina that also undergoes age-related degradation. Accordingly, it is very likely that a panel of simultaneous interventions will be required, each addressing a subset of the many types of dysfunction that aged skin exhibits. Secondly, the skin is in intimate contact with the rest of the body, especially via the circulation, and is therefore adversely affected by the aging of all other tissues. It is difficult to quantify the impact of this as a driver of skin aging, but there are reasons to suspect that it is substantial. Luckily, the specific types of molecular and cellular damage accumulating in all tissues, the skin included, are broadly similar. In all cases they can be classified into just seven major categories. Furthermore, for each such category, it seems likely that all examples within a category will in due course be amenable to repair by broadly the same type of intervention. Two of these seven categories concern extracellular material: accumulation of miscellaneous detritus that has escaped the attention of all mechanisms for degradation or excretion, and biophysical dysfunction caused by chemical or physical alterations to the extracellular matrix. In the skin, the latter is of particular interest in relation to the basal lamina separating the dermis and the epidermis, and also the interstitial extracellular matrix permeating the dermis. These lattices of structural proteins (mostly collagen and elastin) are recycled only slowly, with half-lives of the same order of magnitude as the human lifespan, so they have ample opportunity to accumulate damage. One major type of such damage is random crosslinking, predominantly caused by sequences of reactions between circulating monosaccharides and lysine or arginine residues of ECM proteins. These reactions, collectively referred to as glycation, sometimes result in covalent linkages (especially one particular structure termed glucosepane) between juxtaposed proteins, progressively diminishing the elasticity of the ECM as a whole, with macroscopic consequences in terms of skin vitality. Similar chemistry also results

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in adducts attached to individual amino acids: these probably do not alter the ECMs biomechanical properties, but they may be immunogenic, especially in the increasingly autoimmunity-prone aged environment. In addition, simple rupture of the peptide backbone of ECM proteins will go unrepaired until the protein is subjected to wholesale degradation and replacement, which as noted above is a rare process. This may be a less significant type of ECM damage than in other tissues, such as the walls of the major arteries, but it cannot safely be ignored. Accordingly, there is a strong case for exploring methods to stimulate faster turnover of the skin ECM, thereby to replace it with pristine ECM lacking these modifications. Unfortunately, there is evidence that crosslinking renders ECM somewhat resistant to degradation by proteases naturally secreted by fibroblasts an obstacle that must be overcome if the loss of elasticity just mentioned can be fully reversed. However, after many years of pessimism, there is now new hope for the development of pharmaceuticals capable of cleaving the most abundant glycation-induced crosslinks. Similarly, at the intracellular level there are changes during aging which may contribute substantially to the skins biophysical properties and thus compromise its perceived youth.The most important may be the accumulation in quiescent dermal fibroblasts, typically in the lysosome, of a variety of molecular detritus that is created as byproducts of normal metabolism but is not then degraded or excreted. Pigmented material in particular, whether intracellular or extracellular, constitutes an important aspect of aging. A promising approach to repairing this type of damage is to introduce into cells enzymes (or the genes encoding them), found elsewhere in the biosphere (especially in bacteria), that can degrade such compounds.

It is also necessary to consider aging at the cellular level, i.e. changes in cell number as opposed to cell structure. Thinning of the skin, especially the dermis, contributes greatly to skin aging, and is largely a consequence of depletion of fibroblast density. In the epidermis, stem cell number may also decline. Cell loss is the natural target for stem cell therapies of various means, and especially in relation to the epidermis we are already seeing encouraging progress, not least as a result of the relevance to burns therapy. In sum, a sophisticated multi-component assault will probably be necessary if we are to achieve complete rejuvenation of aged skin but this is no longer a utopian goal. With sufficient resources and determination, it can be achieved within the foreseeable future.

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Leslie Baumann
Dr. Leslie Baumann, Chief Executive Officer, Baumann Cosmetic & Research Institute, Miami, FL, USA
Dr. Leslie Baumann is a dermatologist, researcher, professor and well-known author. In 1997, she chaired the first Division of Cosmetic Dermatology in the USA at the University of Miami. Her textbook Cosmetic Dermatology (McGraw Hill 2002, 2010) was the first textbook on the subject and is currently the bestselling cosmetic dermatology textbook worldwide. She performed the research trials in the USA that led to FDA approval of Botox, Dysport, Sculptra, Juvederm, and many other products and procedures. She has performed clinical research trials for over 50 cosmetic and pharmaceutical companies.

Her New York Times bestselling book The Skin Type Solution is in 5 countries and describes her skin typing system that is based on her years of research on skin care. She authors a skin care blog on Yahoo! Health that is read by millions of people. Dr. Leslie Baumann does not have her own skin care line. Her clinical approach for skin care and cosmetic medicine combines science with practical solutions.

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A Dermatologists View on Skin Rejuvenation

Skin Rejuvenation requires a multifaceted approach. The most successful outcomes occur when the patient is properly educated and motivated, and a proper skin care regimen is combined with in-office procedures.The current trend is to develop at home procedures that mimic in-office procedures. This lecture will discuss the thought process that dermatologists go through when selecting skin care products and cosmetic procedures for facial rejuvenation. The first step in planning a facial rejuvenation plan is evaluating these 4 facial characteristics to decide what cosmetic procedures are best suited for the patient. 1. Evenness of skin color 2. Skin surface texture 3. Presence of wrinkles 4. Facial volume. A youthful face has evenness of color. Visible blood vessels and accumulation of melanin contribute to unevenness of color. Lasers, light devices, skin peels and skin care products are combined to treat uneven skin tone. The theory of photothermolysis and use of light to treat skin discoloration will be discussed. Skin surface texture is determined by the condition of the stratum corneum. Methods used to smooth the stratum corneum include facial scrubs, facial brushes, microdermabrasion and chemical peels. Wrinkles are caused by the loss of collagen, elastin and hyaluronic acid.Topical products have been largely unsuccessful in treating the loss of collagen, elastin and hyaluronic acid with the exception of retinoids. The science of topical retinoids and the use of dermal fillers and botulinum toxin to treat aged skin will be discussed. Facial volume loss is a major cause of skin aging. Stem cells, fat and dermal fillers are being used to correct facial volume defects. The state of the art of stem cells will be discussed. The second step in planning a facial rejuvenation plan is determining the patients Baumann Skin Type using a questionnaire known as the Baumann Skin Type Indicator (BSTI). There are 16 individual Baumann Skin Types based on the combinations of the following parameters: 1. Oily vs dry 2. Sensitive vs resistant 3. Pigmented vs nonpigmented 4. Wrinkled vs non-wrinkled (tight) Combining these 4 parameters gives 16 possible combinations. This lecture will briefly describe the 16 skin types and will focus on the science behind what causes the various skin conditions. A future research aim is to identify the genetic fingerprint of various skin types.

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Carlo Pincelli
Professor of Dermatology, School of Biosciences and Biotechnologies, University of Modena and Reggio Emilia, Modena, Italy
Carlo Pincelli received his training in Dermatology at the University of Modena and Reggio Emilia in Italy and at St. Johns Hospital for Diseases of the Skin in London. He spent two years as a visiting research fellow at the University of California San Francisco in 1986-1988 and 9 months at the Department of Dermatology Boston University in 1994 as a visiting scientist. He became Professor of Clinical Dermatology in 1990. Five years laterhewas appointed as Director of Research of the Laboratory of Cutaneous Biology at the University of Modena and Reggio Emilia.

As epidermal expert, he has brought particular contributions in areas such as stem cells, molecular and cell biology and apoptosis, exploring deeply the pathogenic mechanisms implicated in skin pathologies. He is co-founder and Chief Executive Of-ficer of the academic spinoff Pincell S.r.l.Pr. Pincelli is also co-inventor of two international patents and author of over hundred-fifty articles in peer-reviewed journals. In 2006-2007, he has been President of the European Society for Dermatological Research (ESDR). He is currently member of the Board of Trustees of the European Skin Research Foundation (ESRF).

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Stem cells and skin rejuvenation

Ageing is an inevitable process, with both intrinsic and extrinsic determinants, which involves all tissues and organs of the body and has particular repercussions and evidence in the skin. Intrinsic ageing of the skin occurs as a natural consequence of physiological changes over time at variable rates. Extrinsic factors are, to varying degrees, controllable and include exposure to sunlight, pollution or nicotine, repetitive muscle movements like squinting of frowning, and miscellaneous lifestyle components such as diet, sleeping position and overall health. Skin aging involves increased susceptibility to injury and infection, reduced wound healing, loss of dermal elasticity, poor epidermal barrier maintenance, wrinkling, hair loss, and increased cancer risk. Since birth, skin homeostasis and integrity is guaranteed by the presence of Keratinocyte Stem Cells (KSC) and by the correct balance between proliferation, differentiation and apoptosis. KSCs self-renew and generate the different lineages that form the mature tissue. As we grow old, epidermis becomes more fragile and susceptible to trauma, with an increasing rate of impaired wound healing. The decline of tissue regenerative potential is a hallmark of ageing and may be due to related changes in tissue-specific stem cells. Anyway, given that epidermal functionality must be maintained throughout life, it was tacitly understood that the epidermal stem cell remained functional as long as the organism lived and that this cell population met the continual demand for new cells. Studies in mouse skin confirmed that epidermal stem cells remain functional and appear to resist to cellular ageing. Recent works, demonstrate that it is the Transit amplifying (TA)-cells population whose functionality changes as skin ages and that this change mantains the integrity of the epidermis in old mice. TA cells appear to be more affected than stem keratinocytes by ageing also in human epidermis. KSCs reside in a special microenvironment, the niche, that allows them to maintain their unique features and stemness. KSC are located within the basal layer of epidermis and rest upon the basal membrane that is rich in extracellular matrix and growth factors. As basal keratinocytes exit the niche, they move into suprabasal layers, and different microenviromental stimuli influence their destiny. Under normal conditions, basal keratinocytes leave the basal niche to undergo terminal differrentiation. This critical process is regulated by an increasing number of signals. The niche concept in itself implies that KSCs, in order to maintain longevity and to ensure continue tissue renewal, need to be protected from apoptosis, and all the factors implicated have to be taken into consideration during skin ageing. Some integrin family members are downregulated during skin ageing. Moreover,

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survivin, an Inhibitor of Apoptosis Protein (IAPs) family member, implicated in cell cycle regulation and suppression of apoptosis, is strongly expressed in KSCs and protect these cells from apoptosis induced by UVB radiations. Among other factors, Notch ligands expressed in basal keratinocytes bind to Notch receptors in suprabasal cells, thus promoting the commitment of basal keratinocytes to differentiation. Notch proteins and their signaling pathways seem to be modulated during skin ageing. Finally, neurotrophins and their receptors play an important role in mantainig epidermal homeostasis. Furthermore, dermis plays a critical role in sustaining the stem cell population.The crosstalk between the epidermal and dermal compartment is necessary to maintain epidermal homeostasis. For instance, factors produced in the dermal compartment can act on stem cells. The chronological aging and photoaging of skin is accompanied by an extensive ECM remodelling in the dermis, with a decrease in collagen production and a reduction of skin tone and elasticity, fibrosis and increased risk to develop aggressive and invasive skin cancers.

Therefore, we can conclude that KSCs behaviour is surely age-dependent and strictly related to individual characteristics. However, KSC descendent cells and all the factors involved in KSC niche have to be taken into account in human skin during ageing, in order to plan cosmetical and medical interventions in physiological and pathological conditions.

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Melanie Swan
Futurist and Applied Genomics Expert, MS Futures Group, DIYgenomics, Palo Alto, USA
Melanie Swan is a Research Fellow at DIYgenomics, a non-profit research organization she founded in March 2010. The goal of DIYgenomics is to realize personalized medicine by establishing baseline measures of wellness and custom interventions while conditions are pre-clinical. The organization is engaged in the design and operation of research studies investigating genotype-phenotype linkage and personalized intervention. Ms. Swan has a quantitative risk assessment background and became interested in the predictive risk modeling and practical applications of personal genomics in 2008. Her educational background includes an MBA in Finance and Accounting from the Wharton School of the University of Pennsylvania, a BA in French and

Economics from Georgetown University, and recent coursework in bioscience, nanotechnology, physics, computer science, and philosophy. She is a faculty member at Singularity University and an Affiliate Scholar at the Institute for Ethics and Emerging Technologies. Ms. Swan serves as an advisor to research foundations, government agencies, corporations, and startups and is active in the community promoting science and technology, and opportunities for women.

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Translational reverse-aging research: latest advances

This talk provides a review of some of the most important advances in translational anti-aging skin research, particularly with regard to personalized genomics. Personalized genomics is an emerging field that incorporates the genetic sequencing profiles of individuals into health-related decisions. The field investigates genetic point mutations, structural variation, epigenomics, RNA expression, and microbiome integration. Some current applications of personalized genomics include the assessment of ancestry, carrier status, disease risk, and drug response. Personalized genotyping is carried out through medical professionals and consumer-directed services. Over 100,000 individuals worldwide have subscribed to personal genome services since they became available in late 2007. The validity and utility of disease risk assessment has been criticized as there is variance between services in disease risk interpretation. However drug response is more definitive and accepted. As of July 2011, in the U.S., the Food and Drug Administration has validated genomic biomarkers for approximately 75 drugs. The broader role for personalized genomics is as a component of personalized medicine, using information about an individual to select or optimize preventive or therapeutic care by cohort or individual. Personalized medicine contemplates an extensive range of health and wellness outcomes ranging from cure to improvement to normalization to prevention to enhancement. The generalized hypothesis and methodology of personalized genomic medicine developed by DIYgenomics is that one or more genetic polymorphisms may give rise to out-of-bounds phenotypic biomarkers which may be ameliorated through personalized intervention. For example, the DIYgenomics aging study investigates whether TERT (telomerase reverse transcriptase) mutations may lead to a phenotypic predisposition for shorter-thanaverage telomeres which may be improved through telomerase activation therapy that may also improve skin fitness. Genomic studies are not as well established in antiaging skin research as in general disease risk and drug response research, but are nevertheless surfacing interesting links between genomic polymorphisms and phenotypic conditions. Genomic disease profiling In disease profiling for skin conditions, personalized genomics has been linked to acne, eczema (atopic dermatitis), irritation, erythema, dryness, psoriasis, skin lesions, sun damage, premature aging, xeroderma pigmentosa, development of freckles and solar lentigines, Cockayne syndrome, and Kindler syndrome. In skin cancers, genomic associations have been found in melanoma and cutaneous basal cell carcinoma.

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Genomic links also exist for hair conditions such as hair loss, male pattern baldness, alopecia areata, premature hair graying, and thickness and curliness of hair. Genomic wellness profiling In wellness profiling, personalized genomics may be used to determine an individuals general profile for different kinds of healthiness, a predictive indicator of which conditions may arise over time. Related to skin, areas of genomic wellness profiling include DNA damage repair response, RNA editing capability, immune system response, cancer response, general aging response, and other factors leading to dermis and epidermis wellness. Genomic product response profiling A potential opportunity in personalized genomics is the development of skin care products customized to individuals per genetic profiles. Products could be recommended based on predicted response (efficacy and side effects), and disease risk and wellness profiling. Genomic associations have been found in the skin care product areas of antioxidant treatment, anti-aging DHEA treatment, aluminum powder, and in conjunction with microbiome research, for personalized mosquito repellent. Wrinkle formation: possibly caused by sun damage, ECM degradation, loss of subcutaneous fat, and telomere shortening Wrinkle formation is a central concern of translational anti-aging skin research. There may be multiple causes of wrinkle formation. Sun damage (also known as photodamage, photoaging, and UV-induced damage) is a critical factor. UV-radiation may trigger DNA damage which causes skin to fold and wrinkle. UV-radiation may also trigger the activation of heparanase (an extracellular matrix (ECM) degradation molecule) that increases growth factor interaction between epidermis

and dermis and causes wrinkles. The loss of subcutaneous fat is another important factor in the aging of skin which may contribute to wrinkle formation. Fibrosis is another factor: fibrin (insoluble proteins) in cells becomes infused with deposits over time and turn into fibrosis (thickened connective tissue) which becomes loosened from ECM proteins and triggers wrinkle formation. Solutions might include protease inhibitor drugs, and ACE inhibitors, for example in one study, losartan-treated mice developed less fibrosis than controls. In addition, ECM degradation may be facilitated by telomere shortening which triggers a wound healing phenotype that produces collagen and elastin which degrades the ECM and leads to wrinkle formation. A variety of solutions to impede or reverse wrinkle formation have been suggested in recent research including DNA repair enzymes, the topical application of CoQ10 (to reduce reactive oxygen species (ROS) production and DNA damage triggered by UV-radiation), Vitamin A, and willow bark-derived salicin (activating heat shock proteins (HSPs) which protect cells from stress-induced damage). Retin-A is one of the most widely used skin creams for acne and wrinkles but causes an allergic reaction in a significant percent of individuals. Potential substitutes include retinyl retinoate (a novel hybrid retinoid) to increase the stability of retinol which may have an antiwrinkle effect, and retinyl N-formyl aspartamate, a newly synthesized photostable retinol derivative. Stem cells, regenerative medicine, 3-D skin printing, and skin substitutes Important research is being carried out in stem cells and regenerative medicine, a prominent area for potential near-term health advance. In skin anti-aging,

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the interrelation between stem cell generation, hair follicles, wound healing, and microRNA expression is being investigated. Producing skin for grafts, wound healing, and other purposes is a key application. A number of techniques are being explored including regenerating skin from an individuals own stem cells, 3-D printing of skin with bioscaffolds, and skin substitutes that integrate organic and inorganic material. Personalized genomics is an important emerging field of science being applied to human biology and medicine. Its application in disease risk assessment, wellness profiling, drug response determination, and product response customization may only grow over time and lead to many useful innovations in translational antiaging skin research.

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Since 2008 she has been a Research Manager speciaBio-Science Innovation Manager, lized in anti-ageing and longevity research for LVMH (Mot Hennessy - Louis Vuitton) cosmetic brands. LVMH Recherche, Saint Jean de Braye, France She has conducted over 20 research projects in her Dr. Nizard obtained her PhD in Fundamental and specific areas of interest: protein maintenance, phoApplied Toxicology in 1990 at the French University toageing, heat shock proteins, oxidative stress and Paris Diderot, Paris 7. Three years later she joined the melanogenesis. Research Centre LVMH Recherche as a researcher She has supervised several PhD students and postin the field of skin cells and in 2002 she became a doctoral fellows. She has published more than 20 Bio-Science Innovation Manager. She participated to articles in peer-reviewed journals and she is co-auresearch projects dedicated to innovation for skin care, thor of 13 patents. she developed new models of human skin cell cultures and discovered the pharmacological activity of many new active ingredients.

Carine Nizard

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Skin-cell rejuvenation
In industrialized countries, populations are ageing and over the last 100 years the average life expectancy of people has approximately doubled, a multifactorial phenomenon in which improvement in nutrition and medicine and easier working conditions have played an important role. The idea that ageing is an active continuation of a genetically programmed development of organisms has been partially discredited. Ageing is now admitted to be a loss of equilibrium between the capability of an organism to maintain its repair potential and the frequency and intensity of the damage to which it is exposed. For the skin, the most exposed organ to environmental injuries, free radicals generated by ultraviolet radiation and internal metabolism are considered to be the most important deleterious ageing agents on cellular proteins, lipids, glycans, and DNA-all of which have been extensively reviewed. Therefore antiageing skin care emerged that contained natural freeradical scavengers or technologies focusing upstream with active ingredients which, rather than attack free radicals directly, reinforced the natural cellular proteins that detoxify these free radicals. Recently, sophisticated complementary strategies emerged based on activation of chaperone proteins that protect structures of cellular proteins, and others based on the activation of detoxifying proteins such as proteasome which helps to recycle damaged proteins. Accumulation of oxidized proteins is a hallmark of cellular ageing and is believed to be one of the main contributors to the aged phenotype. This process is dependent on an increased occurrence of oxidative damage to proteins and also on a decreased elimination of oxidatively modified protein. Elimination of oxidized proteins is mainly achieved through degradation by the proteasomal system while certain oxidative protein modifications such as oxidation of methionine residues can be reversed within proteins by the methionine sulfoxide reductases (Msrs) system which represents one of the few repair mechanisms for oxidized proteins. Both systems have been documented to exhibit an age-related decline, notably for the proteasome in human epidermal cells and dermal fibroblasts. UV-irradiation of skin cells is known to induce oxidative stress and to promote protein oxidative damage leading to the formation of carbonyl groups as well as the formation of protein adducts with lipid peroxidation products. However, the fate of such important protein maintenance systems like the proteasome and the Msrs upon UV irradiation has only been recently addressed in skin relevant cellular models. Importantly, a loss in Msr activity would be likely to diminish the antioxidant response, hence

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favouring the accumulation of oxidized proteins, and unfavourably influence keratinocytes recovery from oxidative stress encountered during UV irradiation. Therefore for the proteasome, protecting or stimulating such enzymes responsible for the repair of specific forms of oxidative modification would also be expected to help fight against the accumulation of non functional and potentially harmful oxidized proteins, which represents one of the major deleterious effects associated with intrinsic skin ageing and photo-ageing. Another interesting concept known as hormesis has recently attracted attention in the field of anti-ageing research. The theory behind the approach - that low doses of toxic or harmful substances have a protective effect - is known as hormesis. It makes use of the bodys intrinsic capacity for self-maintenance and repair, by exposing cells and organisms to brief periods of stress. The paradigm for hormesis is exercise, an activity that is both stressful and damaging due to the production of free radicals, acids, stress hormones and tissue damage. However, as an inducer of repair and maintenance processes, the hormetic effect of this strenuous activity has a wide range of health-promoting effects, including slowing down ageing. Limited mitotic life span is observed in many eukaryotic cell types and is interpreted as a manifestation of cellular ageing. Irreversible growth arrest at the G1/S phase of the cell cycle is namely due to the overexpression of cyclin-dependent kinase inhibitors such as p21waf1 and p16ink4a, leading to hypophosphorylation of the retinoblastoma protein. Human diploid fibroblasts (HDFs) in replicative senescence are characterized by a typically enlarged cell shape, senescence-associated b-galactosidase activity, short telomeres and changes in the expression level of many genes. All cells in the organism produce specific molecules - cytokines, which

carry signals, activate the processes of proliferation (dividing), differentiation, apoptosis etc. Through the secretion of various regulatory molecules senescent cells can influence other cells triggering their ageing processes. Recently new active ingredients of natural origin have been proposed to fight the skin cells entry in senescence. LVMH Recherche has performed these 3 skin-cell rejuvenation strategies based on experimental work (in LVMH laboratories or within academic collaborations) with innovative ingredients to develop cosmetic products for LVMH brands. These approaches enable us to have interesting in-vitro skin-cell rejuvenation experimental data and in vivo quantifiable benefits for skin following their cosmetic use.

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Oleg V. Kvitko
Leading scientist, Institute of Genetics and Cytology, National Academy of Sciences, Minsk, Belarus
Oleg V. Kvitko graduated from the biological faculty of the Belarus State University (Minsk) in 1974 and went to the Institute of Genetics and Cytology of the National Academy of Sciences of Belarus (IGC NANB). In 1980, he did a PhD in biology (specialization in genetics) within IGC NANB. In the last 20 years, his research increasingly concentrated on cellular ageing, cancer transformation and stem cells. In these studies he, with his research team, used the informative experimental method based on continuous computer videorecording of microscopic images (computerized videomicroscopy) of living cell cultures revealing unknown processes during aging.

Oleg Kvitko performs an analysis of data and theories in biology of ageing, aiming to explain the causes of ageing and mark ways to elaboration of effective means of slowing ageing and rejuvenation at the levels of a cell and an organism.This resulted in the two new theoretical concepts: participation theory of ageing and developmental theory of rejuvenation. His experimental and theoretical work has been published in over 140 publications. Kvitko is a member of the Belarussian Society of Geneticists and Breeders, a head of Minsk branch of the Gerontological Society of the Russian Academy of Sciences and an associate editor of the journal Theoretical biology and medical modelling.

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Signalling pathways and rejuvenation: risks and opportunities

Finding new effective rejuvenation means can be greatly helped by knowledge of the mechanisms of biological ageing and opposite processes of restoration (repair) of age-related damage. Significant experimental evidence supports the opinion that epigenetic changes underlying unfavourable deviations of gene expression are the major mechanism of ageing. In contrast to the short-termed metabolic alterations of gene expression epigenetic changes causing ageing are relatively stable and lead to disdifferentiation (the term proposed by Richard Cutler), that is the violation of normal physiologically optimal differentiated state of cells. In its turn, disbalance of gene expression leads to the damage in tissues due to accumulation of dysfunctional cells, anomalous proportions of different cell types, disturbances of intercellular communications and deviations from normal cell density. Two molecular mechanisms of epigenetic changes triggering ageing gained a special attention. These are DNA methylation and modifications of chromatin histones (acetylation, phosphorylation and others). Both mechanisms participate in chromatin remodeling and, thereby, regulate gene expression at a global level. In addition to these two processes telomere shortening may be mentioned among the important epigenetic mechanisms of ageing that are perspective targets for rejuvenation. One may object to that because telomere shortening, unlike DNA methylation and histone modifications, may be classified rather as a genetic, not epigenetic process, because it results in changes of a primary nucleotide sequence. However, similarly to conventional epigenetic processes (and in contract to genetic mutations), telomere shortening happens regularly (in every mitotic division of many cell types). In addition, telomere shortening is reversible due to a special enzyme telomerase or other (alternative) mechanisms of telomere elongation. For searching the means of retardation or reversal of aging the possibility of selective epigenetic reprogramming leading to rejuvenation is of special interest. Under this rejuvenating process restoration of telomeric repeats at the ends of chromosomes and physiologically optimal (for a particular cell type) pattern of differential gene activity (determined by DNA cytosine methylation and modifications of chromosomal histone proteins) should take place but dedifferentiation (that happens during somatic cell nuclear transfer into oocyte cytoplasm and construction of induced pluripotent stem cells) should not occur. According to the developmental theory of rejuvenation proposed by the author the natural mechanisms of epigenetic rejuvenation exist but do not always work effeciently enough to withstand the age-related accumulation of epigenetic damage.

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During embryogenesis and postnatal growth the special signalling mechanism reverses epigenetic mistakes and, thereby, rejuvenates cells. This development- and growth-coupled rejuvenating epigenetic repair insures the delay of functional decline to the point of an organisms ability to produce progeny. That is why species with extended growth periods have long lifespans.This rejuvenation mechanism also prevents aging in organisms with negligible senescence. These species have an indeterminate growth (for example, some fishes or mollusks grow throughout their life with little or no senescence, and only an excessive size, not ordinary ageing is a major reason of their death). It appears also that the well known life-extending effect of caloric restriction in rodents (McCay et al., 1935) may be explained by the prolongation of postnatal growth and extension of the work of a growth-coupled rejuvenation process. The natural process of rejuvenation is driven by the same extracellular morphogenetic molecules (morphogenes) that regulate embryogenesis. As a rule, morphogenes bind to specific cell membrane receptors activating signalling pathways inside cells, which stimulate or repress gene expression. There are several families of morphogenes (Wnt, Notch, Hedgehog, FGF, TGF-beta and others). Because of the regulatory pleiotropy (when one molecule takes part in different signalling pathways) morphogenes, simultaneously with performing their canonical morphogenetic functions, stimulate telomerase and trigger restoration of physiologically optimal gene expression by regulation of enzymes that modify DNA (methylation) and chromosomal histone proteins. After the end of development and growth fluctuations of morphogens in tissues decrease (flatten) and become insufficient for initiating the cascades of events resulting in rejuvenation of the whole body. Since human species has a genetically

restricted growth, for counteracting aging and extending human life intensive (embryo-like) production of morphogenes should be reestablished in many or all tissues of an adult organism. At the systemic level stimulation of epigenetic rejuvenation in different parts of the body may be achieved by neuroendocrine regulation. In particular, rejuvenation-promoting modifications of neuroendocrine processes can be reached by special psychological practices. This idea is supported by the recent research on influence of some forms of meditation on telomere shortening and telomerase activity in immune cells (Jacobs et al., 2011). In frames of the developmental theory of rejuvenation a new meditation technique has been proposed developmental meditation (Kvitko, 2009). In addition to systemic methods of rejuvenation, topical influences may give invaluable rejuvenation effects. For example, skin provides unique possibilities for the elaboration and practical application of topical rejuvenation means. In this context it seems reasonable to consider some additional aspects of a possible mechanism of rejuvenation signalling. It is known that Wnt (one of morphogens) can activate the Myc protooncogene which incodes a transcription factor that activates the production of telomerase and regulates expression of 15% of all genes through recruiting histone acetyltransferases that modify chromosomal histones. Thus, extracellular morphogenetic signals may stimulate rejuvenation pathways through protooncogenes. Possible involvement of protooncogenes in rejuvenation signalling should be taken into consideration in cell culture experiments aiming at finding effective and safe regimes of topical application of rejuvenation ingredients. In particular, optimal regimes of using rejuvenation means may be based on

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the concept of transient (reversible) immortalization. This proposition is supported by the experimental evidence on transient immortalization of mammalian cells. Irreversible immortalization is dangerous and inappropriate. Learned from nature safe and effective means of rejuvenation that works in developing and growing organism may exploit nonlinear, pulsed regimes of rejuvenation signalling. Improved methods of long-term (from days to months) uninterrupted computer videomicroscopy of living cell cultures provide unique experimental tools for elaboration of new effective formulas for topical rejuvenation.

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Laure Ritti
Research Investigator, Department of Dermatology Photoaging and Aging Research Program, University of Michigan, Ann Arbor, USA
Dr. Ritti received her Master of Science in Cell Biology in 1996 and her PhD in Biochemistry and Molecular Biology in 2001 from the Universit de Reims Champagne-Ardenne in France. She completed postdoctoral trainings at the Universit de Reims Champagne in the Department of Biochemistry and Molecular Biology, and in the United States, in the Department of Dermatology at the University of Michigan during four years. She has been recruited as Research Investigator to join the Dermatology Department Faculty at the University of Michigan in 2006. She studied specifically

the molecular and cellular basis for impaired wound healing in aged human skin and the effects of aging on follicular stem cell functions in human skin. Her current research focuses on elucidating the mechanisms of altered skin re-epithelialization in aging. Dr. Ritti received an American Skin Association Research Scholar Award in 2007, and a National Institute of Health Research Scientist Development Award four years later. She wrote four book chapters and published dozens of articles in peer-reviewed scientific journals. She also teaches the Basic Science Journal Club to Dermatology Residents at the University of Michigan.

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Controlled Wound Healing for Skin Rejuvenation: Recent progress and Future Challenges
Photoaging defines the premature aging of the skin that primarily results from chronic exposure to ultraviolet (UV) irradiation from the sun. Photoaged skin is characterized by the presence of damaged extracellular matrix components, including type I collagen, the major structural protein in the dermis. Damage to type I collagen is initiated by UV irradiation-induced metalloproteinase activity that degrades collagen fibers into fragments. UV irradiation concomitantly inhibits procollagen production by dermal fibroblasts, thus preventing the dermis to efficiently repair itself. With repeated exposure to UV light overtime, collagen fragmentation accumulates in the skin. The resulting decreased mechanical tension in the dermis is responsible for a self-sustained negative feedback decreasing collagen production in the skin. Thus, damaged collagen cannot be replaced. To reverse this vicious cycle, skin rejuvenation strategies must be aimed at stimulating collagen production in photodamaged skin. Cutaneous wound healing is a regulated process that involves an initial clearing of the wound through an inflammatory phase, followed by tissue restoration including intense deposition of type I collagen and other extracellular matrix proteins. In the past decades, various laser-based devices have been developed to generate a controlled wound healing reaction that ideally triggers minimal inflammation and maximal collagen production in photoaged skin. Providing a detailed analysis of the molecular and cellular alterations induced by several of these lasers, we will show that controlled wound healing is an effective strategy for skin rejuvenation, and will provide a rational for future optimization of treatment strategies and protocols.

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LVMH Recherche Perfumes & Cosmetics

Founded in 1980, LVMH Recherche is a Groupement dIntrt Economique (GIE), whose members are Parfums Christian Dior, Guerlain, Parfums Givenchy and Fresh. LVMH Recherche has about 260 researchers, located in Saint Jean de Braye, Paris, Tokyo and Shanghai. Its mission? To design and develop perfume, fragrance derivatives, skincare products and makeup for several brands of the LVMH Group (Mot Hennessy - LouisVuitton). Our research spans everything from knowledge of skin and cosmetics users to the creation of formulations suitable for strong perceived efficacy and for an international use. Throughout the process, innovation is expressed through the ingredients, formulations and processes that we develop, all united in a single objective: the creation of new beauty solutions inducing emotion of use to our customers.
LVMH Recherche by the numbers: 260 employees More than 1200 products developed per year More than 200 patents More than 50 scientific papers per year Research teams in St Jean de Braye, Paris, Tokyo and Shanghai 36

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