science, technology & beauty 2017 -...

AN EYE ON RESEARCH science, technology & beauty 2017

Exposome

Human Microbiome

SyntheticBiology

Personalised Cosmetics

New scientific fields, rich with the promise of

technology, are emerging, offering avenues

for innovation in a number of sectors: beauty

is not least among them.

What impact does the environment have on

our skin health? Why is our body colonized

by billions of microorganisms? What

purpose do they serve? How can progress

in molecular biology and IT be used to

recreate molecules identical to those that

exist naturally? Is it possible to make tailor-

made cosmetics?

The Scientific Directorate can help to

answer everything you have always

wanted to know but didn’t dare ask…:

exposome, microbiome, synthetic biology

and customised cosmetics and their

applications.

page 4

ExposomeThe environmental impact on the human body

page 14

Human MicrobiomeA new view of the physiology of health and sickness

Synthetic BiologyThe 21st century revolution in technologies and innovation

page 32

Personalised CosmeticsFrom off-the-peg to made-to-measure

page 32page 24

page 14

page 4

Today, genetics is thought to explain less than 25% of the body’s main disorders, with the majority caused by environmental, i.e. non-genetic factors.

Several types of exposure

The types of exposure the human body faces can be placed in one of three broad categories: internal, general external and specific external.

Internal exposure: metabolism, hormonal status, body shape, physical activity, intestinal microflora, inflammatory status, oxidative stress and ageing.

General external exposure: education, social status, mental stress, urban or rural environment and climate.

Specific external exposure: chemical contaminants, pollution, infectious agents, radioactivity, tobacco use, alcoholism, work, diet and sleep.It is important not just to consider the type of exposure but also how it varies over time.

In 2014, G.W. Miller and D.P. Jones2 explained the concept of the exposome, including the

body’s response to environmental factors and endogenous metabolic processes that can impair or alter the chemical substances to which the human body is exposed.

Our genetic heritage programs the course of our life but the influence of

external factors means we rarely realise our full genetic potential.

Many environmental influences are beneficial to our health and well-

being, such as healthy food, education and social relationships, while

others, such as malnutrition, pollution and poverty are harmful.

t

THE ENVIRONMENTAL IMPACT ON THE HUMAN BODY

t

EXPOSOME

In 2005, Dr Christopher P. Wild, Director of the International Agency for Research on Cancer (a WHO agency), put forward a new concept: the exposome. This refers to all the exposures an individual is subjected to from conception via their development in the womb to death and thus supplements the effect of the genome1.

The exposome becomes the cumulative measure of environmental factors and the biological responses associated with them throughout our lives, including exposures linked to the environment, diet, behaviour and endogenous processes.

4L’Oréal 2017 - Research & Innovation / Exposome

Identifying environmental factors and

measuring their impact on the integrity

of the skin in order to prevent and repair

the signs of skin ageing are key aims of

L’Oréal researchers.

5

From understanding the exposome to preventing ageing in healthy skin

Skin constitutes the interface between the outside world and the inside of our bodies. Because it interacts directly with the environment, it is the first target for exposure to environmental stress factors. Its integrity is also linked to our bodies’ good health. As a result, numerous environmental and lifestyle factors have a significant impact on the skin’s appearance and contribute to its ageing.

Sun exposure is the main damage caused to the skin. Pollution, extreme temperatures and ozone are other external factors that contribute to the extrinsic ageing of our skin surface. Lifestyle – sleep, tobacco and alcohol

consumption, physical exercise, etc. – also influences the appearance of the skin.

These various exposure factors generally act in combination and are likely to result in cumulative negative effects. They contribute to the “silent ageing” process of the skin.

The issues involved in studying the exposome are mainly health-related (public health, general toxicology and occupational

medicine), ecological (eco-epidemiology, etc.) or concerned with scientific knowledge (the internal chemical environment).

L’Oréal 2017 - Research & Innovation / Exposome

EXPOSOME HUMAN MICROBIOME

SYNTHETIC BIOLOGY

PERSONNALISED COSMETIC

SO

MM

AIR

E

For decades, L’Oréal’s Research team has been studying skin ageing and in particular, the role of extrinsic, environmental factors3, which act in addition to intrinsic ageing. UV radiation is the most important environmental factor affecting the physiology of the skin. The skin’s exposure to UV radiation has both short and long-term consequences, including erythema (or sunburn) to photoageing, photo-immunosuppression and skin cancers.

Ultraviolet radiation

The solar radiation that reaches the Earth is made up of 50% visible light, 40% infrared and 10% ultraviolet (UV). Each type of radiation is associated with a particular wavelength. The shorter the wavelength, the higher the energy level of the radiation, which is also expressed in terms of power, where the most powerful rays are those with the shortest wavelength. UVB accounts for 0.5% of solar radiation (wavelength 280-320 nm) and UVA for around 9.5% (wavelength 320-400 nm). UVC does not reach the earth because it is absorbed by the ozone layer.

UVBUVB radiation consists of very high-energy rays that penetrate into the deep layers of the epidermis. A small proportion reaches the upper part of the dermis. UVB rays are responsible for sunburn, cell lesions in the epidermis called “sunburn” cells – where the cells enter into a process of programmed cell death, tanning – by stimulating the production of melanin by melanocytes, but also benign and malignant skin lesions (cancers).UVB rays also have beneficial effects. They lead to the production of several small antimicrobial molecules and previtamin D and produce reactions that protect the skin from future exposures. Indeed, stimulating melanin synthesis by melanocytes produces a protective effect, since melanin absorbs a proportion of radiation. This natural photoprotection, which blocks and filters UVB

rays, is still only partial: some UVB radiation still reaches the upper dermis. Moreover, it varies between individuals. People with pale skin, whose epidermis contains melanin that is less effective for dealing with UV radiation, are more sensitive to sunburn and more likely to develop malign skin lesions.

UVA The proportion of UVA radiation produced by the sun and which reaches the surface of the earth is 20 times higher than for UVB. Made up of short (320-340 nm) and long (340-400 nm) UVA rays, it is not stopped by the ozone layer and passes through clouds and glass. UVA rays are produced all day and all year long, regardless of latitude or season, and can penetrate deep into the skin as far as the lower dermis. 80% of UVA rays reach the upper part of the dermis, with 20% penetrating more deeply. Natural photoprotection is inadequate for blocking and filtering them.

Impact of daily exposure to non-extreme solar radiation

A limited proportion of the world’s population is exposed daily to intense solar radiation, the clinical, photoageing and skin cancer-related consequences of which are now known and indisputable. The issue is finding out the consequences of daily exposure to sunlight, which clearly does not cause any clinical symptoms in the short term.

Long UVA rays are the most significant part of the UVA spectrum and they pen-etrate the skin most deeply4. They play a decisive role in many aspects: photoag-eing (their biological effects are barely visible in the short term). DNA damaging through the production of free radicals, which harm all cell types), immune system responses and various photodermatoses (sun-related intolerances and allergies).

t

THE IMPACT OF ENVIRONMENTAL FACTORS ON HEALTHY SKIN


L’Oréal researchers have been investigating this for several years now and have recently produced a summary of the results of their own research and the findings of other teams working in this area in an international scientific journal5.

The standard UV spectrum of daylight has been defined6: it represents non-extreme exposure to the sun, not at the zenith, with a UVA/UVB ratio of 27. This type of exposure does not produce any visible effects (erythema) in people who spend time outdoors, but it can have harmful effects in skin tissue.Daily exposure of this kind was simulated in the laboratory using a device that mimics precisely the spectrum of daily UV radiation.

Clinical effects were observed after repeated daily exposure for one month. These consist of changes to the skin surface, with an increase in pigmentation, and deterioration in the hydration and microtopography of the skin.

At the biological level, there is a reduction in the thickness of the epidermis and an impact on the Langerhans cells responsible for the skin’s immunity, which decline in number, and keratinocytes, which become more prolific. The size of melanocytes increases. The extracellular matrix of the dermis is also affected, with a reduction in the quantity of fibrillin, procollagen I and glycosaminoglycans. There is also an increase in oxidative stress, an accumulation of p53 – the protein responsible for programmed cell death, the generation of inflammatory molecules and damage to cell DNA. In vitro studies have shown that daily radiation leads to an increase in the expression of genes involved in oxidative stress, inflammation and the deterioration of collagen fibres in both compartments of the skin, the epidermis and dermis7.

Most effects appear only after nine exposures, without causing any erythema, at a dose corresponding to 5% of the UV received in Paris during a day in April.

“We don’t age all year round”: two-photon microscopy based evidence

Two-photon microscopy is a high-performance, non-invasive imaging tool. It can be used to study skin tissue in vivo.

L’Oréal Research has carried out a study in real-life conditions designed to observe changes over a year in the density of elastic and collagen fibres in the upper dermis on the dorsal face of the forearm, an area that is protected by clothing in autumn and winter and photo-exposed during the spring and summer. These parameters were correlated to the thickness of the epidermis and density of melanin, and to measurements of skin elasticity and thickness.

The results of the year-long study on female volunteers aged 55 to 65 showed a seasonal effect on the various parameters studied:

The average thickness of the skin (epidermis and dermis) varies over the year, decreasing after the spring (June) and increasing after the summer (September).

The average density of melanin measured changes with the seasons, peaking at the end of the summer (September) and declining in winter. The increase in melanin density is a consequence of photo-induced pigmentation.

Finally, the density of collagen and elastic fibres reduces by 10% between March and June, remains stable between June and September and then increases by 25% for collagen fibres and 17% for elastic fibres between September and March of the following year.

This in vivo study shows that the skin is capable of restoring key parts of the dermis that have deteriorated if it is protected daily, in the case of this study by wearing clothing.

UV and show that even without a visible reaction on the surface of the skin, this exposure leads to clinical and biological impairments of the epidermis and dermis. UVA radiation contributes in large part to the biological effects observed.

These results highlight the impact of chronic exposure to non-extreme doses of



SYNTHETIC BIOLOGY


SO

MM

AIR

E


The catalytic effect of pollution

More and more epidemiological studies show that exposure to air particles contributes to premature skin ageing and that exposure to high concentrations of ozone aggravates pre-existing dermatological conditions.Pollution aggravates skin damage related to UV exposure. Chronic exposure to the particles emitted by vehicle traffic is significantly associated with premature skin ageing, particularly the formation of skin blemishes8. While pigmentation disorders are mainly caused by cumulative UV radiation, their formation is also the result of chronic exposure to particles and gaseous elements in atmospheric pollution.

L’Oréal researchers have conducted two multi-centre clinical studies, one in Mexico and the other in China, in order to evaluate the impact of urban pollution on the skin. They compared the exposure to pollution of individuals, men and women living in areas considered to be polluted compared with less polluted areas (Mexico City/Cuernavaca9 and Shanghai/Chong Ming10).

The results of both studies led to the same conclusions: exposure to atmospheric pollution

causes biological and clinical effects, with no difference between men and women. These effects contribute to skin ageing.

The production of sebum is increased and its composition is modified when the environment is polluted. Parameters associated with the oxidation process are altered, in particular a depletion of vitamin E, a reduction in squalene (a lipid produced by the sebaceous gland) and an increase in oxidized proteins. The barrier function is altered, with a reduction in hydration and chymotrypsine-like activity. Clinical evaluation by dermatologists shows that the skin of individuals in a polluted atmosphere is more susceptible to dryness, eczema and erythema.

Moreover, L’Oréal researchers have shown, for the first time, that oxidized squalene is a marker of environmental pollution11. Squalene represents 15% of the components of sebum. In tests, ozone, long UVA rays and cigarette smoke were shown to be powerful oxidation factors for squalene. The formation of blackheads, the contribution to the appearance of inflammatory acne and changes in skin topography (wrinkles) may be consequences of oxidation.

The exposome therefore underlies the silent ageing of the skin, which can be tackled by protecting the skin from daily attack from UV radiation. This protection enables the skin, which no longer has to defend itself from aggression, to trigger the biological processes necessary for remodelling the fibres in the dermis.

9

Melanin Fibers

3D-Density on

forearm (%)

10

20

30

40

50Sept

(M6)

March

(M12)

March MarchJune September

L’Oréal 2017 - Research & Innovation / Exposome


SYNTHETIC BIOLOGY


SO

MM

AIR

E

Evolution of the density of collagen and elastic fibers

Collagen Fibers 3D-Density on forearm (%) Collagen Fibers 3D-Density on forearm (%)

Sept

(M6)

March

(M6)March

(M12)

March

(M12)

20

40

60

80

100

20

40

60

80

100

The role of tobacco

Each time someone inhales cigarette smoke, over 4,000 dangerous substances enter their lungs. The ageing of their body, face, gums and skin accelerates faster as a result.The harmful effects of tobacco are attributed to the substances found in smoke during combustion: nicotine, carbon monoxide, tar, formaldehyde, hydrogen cyanide, ammonia, mercury, lead and cadmium. These chemical substances limit blood flow to the skin, which leads to a deterioration of support structures such as collagen and elastin. At the biological level, tobacco works in a similar way to UV rays, by activating MMP1 metalloproteins. This produces an imbalance between oxidants and antioxidants, which modifies the activity of melanocytes. Like the particles found in atmospheric pollution, the components of cigarette smoke activate the aryl hydrocarbon receptor (AhR).

These biological effects are manifested at the clinical level by a negative effect on healing due to the reduction in blood flow, higher melanin indices and skin that appears greyer and more wrinkled than that of non-smokers.

Influence of the climate and seasons

The skin acts as a biosensor and adapts to changes in the environment to maintain its internal balance. Its responses can, however, impair its integrity and damage its barrier function. Changes in the atmospheric humidity level can, for example, play a role in exacerbating skin disorders such as psoriasis or atopic dermatitis (skin dryness).

The skin’s normal average temperature is around 33°C. When skin is exposed to cold, repair of its barrier function slows down, the intracellular lipids content of the horny layer reduces and atopic dermatitis is exacerbated. Heat is an environmental factor that contributes


to skin ageing, a phenomenon known as “thermal ageing of the skin”. Exposure to heat contributes to an accumulation of deteriorated elastic fibres in photo-aged skin. Significant ageing of the forearms, which are in contact with the heat from ovens, can be observed in bakers, as well as on the faces of glassblowers.

L’Oréal researchers have carried out two clinical studies in China to determine the influence of the seasons on the appearance of skin. In one, they showed a seasonal darkening of the skin of Chinese women12. This was observed between the months of January and July and can be avoided by applying an SPF 50+ PPD 18 sun care product. In the long term, use of a sunscreen is thought to help reduce pigmentation disorders linked to exposure to UVA and UVB.In the second study13, they showed that signs of skin ageing such as wrinkles and sagging skin are not affected by the season, winter or summer, but that certain functional parameters were modified in the summer, with an increase in the secretion of sebum, the colour and hydration of the skin, and the melanin content of areas of pigmentation.

The impact of nutrition

Skin symptoms such as dermatitis, stomatitis (inflammation at the corner of the lips), alopecia and loss of pigmentation observed as a result of certain nutritional deficiencies reveal a link between nutrition and skin condition14. In addition, dietary factors and nutritional supplements are likely to influence skin ageing. A diet that is high in antioxidants can slow the effects of ageing and twins who avoid excessive alcohol consumption are perceived as younger. Other studies show that consuming a high level of vegetables and olive oil protects the skin from sun damage, while consuming meat and dairy products is thought to have the opposite effect. It should be noted, however, that some epidemiological studies designed to demonstrate the role of dietary supplements on health have shown negative effects from these supplements. In particular, the SU.VI.MAX study showed that taking an antioxidant gel capsule (120 mg vitamin C, 30 mg

vitamin E, 6 mg beta-carotene, 100 mg selenium and 20 mg zinc) caused a higher incidence of melanoma in women15. The best strategy for neutralising pro-oxidant agents is therefore to eat fruit and vegetables, with supplements only being used in cases of deficiency.

Impact of sleep

The impact of a lack of sleep is associated with an increased risk of numerous chronic diseases, including hypertension, diabetes, obesity and cardiovascular diseases. Its effects on the skin are physical, affecting the aesthetic appearance of the face. Subjects who are lacking in sleep look less attractive and more tired, with changes to several parameters relating to skin colour. Another study shows that lack of sleep affects the eyes (which become red and swollen, with dark circles), eyelids (which sag) and skin, which appears pale and more wrinkled16.

Cosmetics and the exposome

Cosmetics participate in the exposome in the same ways as food and pharmaceutical products. They differ from the environmental factors listed above because the ingredients used in them have demonstrated their ability to reduce or prevent skin ageing in clinical studies and because the safety of the raw materials and finished product has been assessed.

Targeting the exposome and slowing ageing: towards personalised approaches

Our lifestyle increases our environmental exposure and as a consequence, premature ageing. Daily exposure to long UVA radiation and pollution are key factors in premature skin ageing. The immediately visible sign of pigmentation is followed by changes in the texture of the skin and finally, the appearance of wrinkles. Research results are paving the way to routines and devices that adapt to our personal lifestyle, skin type and habits. Sensors measure our environmental stress (via a UV patch), devices produce biological markers to record how our skin reacts and algorithms tell us how to keep our skin healthier for longer and slow down the visible clinical signs of ageing.


March MarchJune SeptemberMarch MarchJune September


SYNTHETIC BIOLOGY


SO

MM

AIR

E


1 Wild, CP. “Complementing the genome with an “exposome”: the outstanding challenge of environmental exposure measurement in molecular epidemiology”. Cancer Epidemiol Biomarkers Prev. 2005 Aug;14(8):1847-50

2 Miller G W, Jones DP. (January 2014). The Nature of Nurture: Refining the Definition of the Exposome. Toxicological Sciences 137 (1): 1–2.

3 Krutmann J, Bouloc A, Sore G, Bernard BA, Passeron T. The skin aging exposome. J Dermatol Sci. 2016 Sep 28. pii: S0923-1811(16)30816-7. doi: 10.1016/j.jdermsci.2016.09.015. Review.

4 Bernerd F, Asselineau D. UVA exposure of human skin reconstructed in vitro induces apoptosis of dermal fibroblasts: subsequent connective tissue repair and implications in photoaging. Cell. Death Differ. 1998, 5, 792–802.

5 Marionnet C, Tricaud C, Bernerd F. Exposure to Non-Extreme Solar UV Daylight: Spectral Characterization, Effects on Skin and Photoprotection. Int. J. Mol. Sci. 2015, 16, 68-90; doi:10.3390/ijms16010068

6 Christiaens FJ, Chardon A, Fourtanier A, Frederick J.E. Standard ultraviolet daylight for non-extreme exposure conditions. Photochem. Photobiol. 2005, 81, 874–878.

7 Marionnet C, Pierrard C, Lejeune F, Sok J, Thomas M, Bernerd F. Different oxidative stress response in keratinocytes and fibroblasts of reconstructed skin exposed to non-extreme daily-ultraviolet radiation. PLoS One 2010, 5, e12059.

8 Krutmann J, Liu W, Li L, Pan X, Crawford M, Sore G, Seite S. Pollution and skin: From epidemiological and mechanistic studies to clinical implications. J Dermatol Sci. 2014 Sep 16. pii: S0923- 1811(14)00193-5. doi: 10.1016/j.jdermsci.2014.08.008

9 Lefebvre MA, Pham DM, Boussouira B, Bernard D, Camus C, Nguyen QL. Evaluation of the impact of urban pollution on the quality of skin: a multicentre study in Mexico. Int J Cosmet Sci. 2015 Jun;37(3):329-38. doi: 10.1111/ics.12203.

10 Lefebvre MA, Pham DM, Boussouira B, Qiu H3 Ye C, Long X, Chen R, Gu W, Laurent A, Nguyen Q. Consequences of urban pollution upon skin status. A controlled study in Shanghai area. Int J Cosmet Sci. 2016 Jun;38(3):217-23. doi: 10.1111/ics.12270.

11 Pham DM, Boussouira B, Moyal D, Nguyen Q. Oxidization of squalene, a human skin lipid: a new and reliable marker of environmental pollution studies. Int J Cosmet Sci. 2015 Aug;37(4):357-65. doi: 10.1111/ics.12208.

12 Qiu H, Flament F, Long X, Wu J, Xu M, Leger DS, Meaudre H, Senee J, Piot B, Bazin R. Seasonal skin darkening in Chinese women: the Shanghaiese experience of daily sun protection. Clin Cosmet Investig Dermatol. 2013 May 31;6:151-8. doi: 10.2147/CCID.S41578.

13 Qiu H, Long X, Ye JC, Hou J, Senee J, Laurent A, Bazin R, Flament F, Adam A, Coutet J, Piot B. Influence of season on some skin properties: winter vs. summer, as experienced by 354 Shanghainese women of various ages. Int J Cosmet Sci. 2011 Aug;33(4):377-83. doi: 10.1111/j.1468- 2494.2011.00639.x.

14 Pezdirc K, Hutchesson M, Whitehead R, Ozakinci G, Perrett D, Collins CE. Can dietary intake influence perception of and measured appearance? A systematic review. Nutr. Res. 35 (3) (2015) 175– 197.

15 Hercberg S, Ezzedine K, Guinot C, Preziosi P, Galan P, Bertrais S, Estaquio S, Briançon S, Favier A, Latreille J, Malvy D. Antioxidant supplementation increases the risk of skin cancers in women but not in men. J. Nutr. 137 (9) (2007) 2098–2105.

16 Sundelin T, Lekander M, Kecklund G, Van Someren EJ, Olsson A, AxelssonJ. Cues of fatigue: effects of sleep deprivation on facial appearance. Sleep 36 (9) (2013) 1355–1360.

REFERENCES


t


SYNTHETIC BIOLOGY


SO

MM

AIR

E

According to the view of an increasing number of biologists, a human being is no longer sim-ply an individual derived from a fertilised egg containing the genes of their father and mother. They are part of a complex ecosystem, which also consists of the billions of microbial species that live in areas of their bodies as diverse as

the nose, mouth, skin, intestines and lungs, all of which are in contact with the outside world.

These microorganisms are bacteria, fun-gi, viruses and archae bacteria (organisms formed of a single cell with no nucleus). They live in communities which either cooperate or

t

In recent years, the human microbiome – the billions of microorganisms

that live on and inside our bodies – has become a huge field of research

and development for the agrofood, pharmaceutical and biotech

industries, as well as the cosmetics sector.

HUMAN MICROBIOME

t

A NEW VIEW OF THE PHYSIOLOGY OF HEALTH AND SICKNESS

14L’Oréal 2017 - Research & Innovation / Human Microbiome


Maintaining or caring for your body is

about acting on these communities

of microorganisms by rebalancing

a particular flora or modulating the

interactions between microbes, or

between microbes and the host.

compete with each other. Keeping them in bal-ance is necessary to maintain our health and well-being. These communities perform several functions that are essential to health: they train our im-mune system to recognise foreign organisms and protect our body from invasion by patho-genic agents. They produce essential vitamins and enzymes needed for digestion, and syn-thesise anti-inflammatory molecules.

Our microbiome is personal to us, although most of the microbial genes are widely shared

in a given population. The diversity of these communities between individuals is high in re-lation to the skin and intestines but low in saliva and the vagina.

The microbiome adapts in response to chang-es such as diet, taking antibiotics, the envi-ronment and age, i.e. throughout an individu-al’s lifetime. These changes are likely to lead to an imbalance that causes disease. Some skin disorders, obesity and type-2 diabetes are thought to be caused by an unbalanced mi-crobiome.

The challenge researchers face today is under-standing how microbiomes change, and iden-tifying and evaluating the extent to which ge-netic and environmental factors are involved in microbiome variations. The aim is to find ways of preventing variations and/or rebalancing communities of microorganisms.

The prerequisite for these studies is knowing what constitutes a healthy microbiome. The wide diversity of microbial communities, with 1,000 different species together accounting for 300 times more genes than all our human cells, can now be understood thanks to the considerable progress made in high-speed genetic sequencing. These technologies have also paved the way to an understanding of the interactions between microorganisms and the human body.

In future, we will see the development of new

diagnostic approaches based on the charac-teristics of the microbiome, along with new diets and treatment strategies to correct imbalances. Microbiome balance and diversity will be checked regularly using diagnostic tests. Treatments will be personalised.


SYNTHETIC BIOLOGY


SO

MM

AIR

E

The microorganisms that live in our intestine perform a number of functions: for example, they are involved in digesting food, produce certain vitamins (B and K) and anti-inflammatory molecules, and train the immune system to distinguish between benign and harmful elements.

The intestinal microbiome also has an effect on the development and operation of the brain. Repeated exposure to antibiotics, inappropriate diets and age can all cause an imbalance in the microbiome. Its influence on human physiology begins even before birth and continues through to old age. A vaginal delivery

The intestine, with a surface area of approximately 250 m2 (the size of

a tennis court) is the largest area of interaction between the inside of

our body and the environment. The 100 million neurones it contains have

earned it the name of ‘second brain’. Because it is constantly exposed

to foreign organisms, its microbiome contains by far the largest number

of cells in our body, to the extent that it has been described as an organ

in its own right, which looks after our health and well-being throughout

our lifetime.

10,000 billion bacteria, 10 times more than all our cells combined, 1,000 different species and a weight of 2 kg.

t

THE INTESTINAL MICROBIOME: AN ORGAN IN ITS OWN RIGHT


provides the newborn with seed bacteria. These help it to digest breast milk, which contains complex sugars to feed the bacteria in the newborn’s intestine, which are essential for the development of its immune system and brain. In elderly people, inadequate chewing of food and/or minimal saliva production can result in pathogenic agents getting into the intestinal tract and create an imbalance in the intestinal microbiome.

Genetic traits are no longer solely responsible for disease. It would even seem that environmental factors and an imbalance between microbial communities are more important than genetics. Crohn’s disease,

ulcerative colitis, obesity, type-2 diabetes and colorectal cancer but also allergies, autism and depression are chronic conditions that are now being linked to an imbalance in the intestinal ecosystem.

The intestine and skin are major sites for immunological surveillance and studies show a link between certain intestinal disorders and skin infections: for example, a third of patients suffering from Crohn’s disease also have skin lesions caused by psoriasis and intestinal infections caused by the bacterium Helicobacter pylori are associated with rosacea.

Numerous questions arise as a result: how do these microorganisms determine the quality, appearance and beauty of our skin? What is their role in the ageing process, and how do they react with the environment? What is the role of the chemical substances produced by these microorganisms on the surface of the skin (in relation to the environment, UV radiation and pollution)? Moreover, what role do these microorganisms play in the appearance of odours associated with heat, age, etc.?

Figures

The skin, with a surface area of around 1.8 m2, acts as a barrier between our body and the environment. The epidermis renews itself constantly, from the base layer up to the outermost surface layer, which is formed of totally different cells called corneocytes. Each cm2 of skin is home to 1 million bacteria.

Over 500 species of bacteria, potentially expressing over 2 million genes, have been identified on healthy human skin to date.

The diversity of skin microbiomes

The various habitats of skin microorganisms are determined by the thickness of the skin, folds, and the density of hair follicles and sweat and sebaceous glands.

Age, gender and the environment also contribute to the variability of the microflora. In utero, foetal skin is sterile, but it begins to be colonised by microorganisms within a few minutes of birth. A newborn’s microbiome is similar all over the skin surface but diversifies as the various regions of the skin develop their own characteristics in terms of humidity, temperature, pH or sebum.

Following the discovery of the microbiome, it is no longer the stratum

corneum, the outermost layer of the epidermis, that forms the interface

between the inside and outside of the body but a living ecosystem made

up of billions of microorganisms.

t

THE SKIN MICROBIOME



SYNTHETIC BIOLOGY


SO

MM

AIR

E

Impact of the environment and beauty routines on the microbiome

Researchers have produced maps of bacteria and chemical molecules present on the skin of individuals to evaluate the impact of the environment on the microbiome2. Environmental factors that are specific to the individual, such as where they live, their exposure to UV radiation and pollution, their profession, clothing, diet and any medical treatment they may be taking (particularly antibiotics) can affect how their skin is colonised by bacteria. Cleansers, moisturisers and other products applied to the skin are potential factors that contribute to variations in the skin microbiome.

More diversity between skin sites than individuals

There is less diversity between individuals from different ethnic groups for a given skin site than between two sites on the same individual. A study carried out by L’Oréal compared the microbiomes of three skin sites – the forearm, armpit and scalp – of men from six ethnic groups living in New York3. The results showed that ethnicity is a secondary factor in determining the composition of the skin microbiome compared with the area of the body studied.

(Gb) Glabella

(Ai) Alar crease

(Ea) External auditory canal

(Tw) Toe web space Front Back

(Um) Umbilicus

(Ic) Inguinal crease

(Hp) Hypothenar palm

(Id) Interdigital web space

(Vf) Volar foream

(Ac) Antecubital fossa

(Ax) Axilary vault

(Mb) Manubrium

(Na) Nadre

Plantar heel (Ph)

Poplitea fossa (Pc)l

Gluteal crease (Gc)

Buttock (Bt)

Back (Ba)

Occiput (Oc)

Retroauricular crease (Ra)

ActinobacteriaCorynebacterineaePropionibacterineaeMicrococcineaeOther actinobacteria

BacteroidetesCyanobacteria

FirmicutesOther firmicutesStaphylococcaceae

ProteobacteriaDivisions contributing < 1%Unclassified

Three bacterial communities have been identified on the skin: those that colonise damp areas (in green), those that prefer dry areas (in red) and those that inhabit areas that are high in sebum (in blue). Source: Grice et al, 20091


Maintaining the resident microflora is essential for keeping skin healthy

Two kinds of flora coexist on the skin: the resident flora, viewed as the commensal flora for the skin microbiome, which are relatively stable in terms of quantity and distribution, and the transient flora, made up of microorganisms that come from the environment. Commensal bacteria are closely linked to skin health. Symbiotic interactions – close, specific associations, which are beneficial to all parties – between these bacteria and the skin organise and modulate the host’s innate immune response.

Skin infections can occur when the diversity of bacterial communities diminishes and there is a change in the interactions between microorganisms and the host

Characterising the microbiome produces a microbiological signature for healthy and unhealthy skin. Used during or after treatment, it provides a means of evaluating its effectiveness in terms of rebalancing microbial

communities. Dermatological infections are often distinguished by less diverse microbial communities around the damaged areas.

There is an increasing number of publications on characterising the microbiome in skin infections. Most of the examples selected below have come from work carried out in L’Oréal’s Research laboratories.

It has recently been shown that the microbiome is altered in affected areas of skin among people suffering from vitiligo or atopic dermatitis, that the imbalance in the proportions of microorganisms that dominate the scalp is correlated with the presence of dandruff and that the unpleasant smell of armpits is due to an increased quantity of bacteria in this area. Clinical studies have proven that using an emollient can modify the microbiome of atopic skin and move it closer to the microbiome of healthy skin, and that extracts of the bacterium Vitreoscilla filiformis found in thermal waters or some probiotics have a therapeutic effect on some dermatological infections.



SYNTHETIC BIOLOGY


SO

MM

AIR

E

Alteration of microbial networks in skin affected by vitiligo

Vitiligo is a chronic skin infection characterised by a localised loss of skin pigmentation due to the death of melanocytes. The loss of melanocytes could be caused by several factors, in particular an autoimmune attack by the pigment cells themselves.

Studying the microbiome of subjects affected by vitiligo showed that the diversity of the microbial community diminishes in affected (depigmented) areas compared with unaffected areas and that the interactions between the different members of the bacterial community in the unaffected areas are different and more significant than in the affected areas. We do not know, however, whether the change in the microbiome is a cause or a consequence

of an alteration in physiology of the skin in the depigmented areas4.

Scalp and dandruff microbiome

L’Oréal researchers have shown that dandruff is associated with an imbalance in the proportion of the main bacterial and fungal populations on the scalp.Comparing the microbiome of the scalp of individuals suffering from dandruff and that of a control population reveals that the presence of dandruff is correlated with a higher presence of the yeast Malassezia restricta and Staphylococcus epidermidis and a lower presence of Propionibacterium acnes. These differences are also observed between dandruff and non-dandruff areas in individuals affected by dandruff. The number of Malassezia restricta is around 10 times higher in the affected areas

“Good Guys”

Staphylococcus epidermidis Staphylococcus aureus

HEALTHY SKIN SKIN INFECTION

Immune Response Immunosuppression

“Bad Guys”

Resident commensal microorganisms such as Staphylococcus epidermis, which act as the so-called

‘good guys’, protect the skin by activating an immune response, while pathogenic microorganisms such

as Staphylococcus aureus (golden staph), the ‘bad guys’, cause immunosuppression and result in skin

infections.


compared with dandruff-free areas. The results of the first study carried out in France5 have been confirmed in China6 and Brazil7.

Until now, anti-dandruff treatment consisted of targeting the yeast Malassezia restricta using antifungal agents. Since the imbalance in the proportion of microbial populations that dominate the scalp was discovered, strategies have been developed to re-establish the composition of the microbiome on scalps not affected by dandruff in areas that are affected.

The microbiome and skin infections: atopic dermatitis or eczema

Atopic dermatitis (or atopic eczema) is a chronic, pruriginous and inflammatory dermatosis, with periodic outbreaks. It is the most common form of dermatosis among children, affecting 15 to 20% of children under the age of seven. The prescribed treatments – topical dermocorticoids – do not cure the condition but help patients to manage outbreaks. The lesions caused have long been associated with colonisation by an opportunistic pathogen, golden staph, which sometimes results in antimicrobial treatment.

An analysis of the skin microflora of subjects affected by atopy or not, showed that the skin microbiome of atopic patients is different from that of a healthy subject and that the microbiome of the affected areas is significantly different from adjacent unaffected areas8. Bacterial diversity in atopic patients is less significant than in healthy subjects and this diversity is lower in affected areas than in adjacent unaffected areas. Limited bacterial diversity on the skin is therefore a marker associated with atopy. Increasing microbial diversity would help to protect against the marked inflammatory outbreaks observed in atopic patients.

After applying an emollient (Lipikar Baume AP, La Roche-Posay) twice a day (morning and night) for three months, the bacterial community in the affected areas moved towards of the adjacent unaffected areas. This study shows that it is possible to use the skin’s microbiological signature to diagnose a patient’s atopic status. Early changes in the

composition of the skin microbiome provide a non-invasive means of predicting the risks of a relapse and preventing chronic lesions, which are very disabling for patients, by providing the most appropriate treatment solutions.

Vitreoscilla filiformis: a bacterium that contributes to the integrity of the skin

Several studies have been carried out by L’Oréal researchers to understand the effect of Vitreoscilla filiformis – a bacterium that lives in thermal water that is known for its dermatological benefits – on atopic dermatitis and its beneficial effects on the skin. The results show that compared with a placebo, applying a cream containing a 5% lysate of V. filiformis significantly reduced signs of atopic dermatitis and pruritus9. Another study has shown that V. filiformis improves seborrheic dermatitis10. More generally, treatments using bacteria or bacterial extracts are increasingly being considered as a way of rebalancing flora, to produce therapeutic molecules and to modulate interactions between microorganisms, and between microorganisms and the host, with biological peptides.

Probiotics strengthen the defence mechanisms of reactive skin

Reactive skins are highly sensitive to physical or chemical irritants. Their barrier function is damaged, increasing the penetration of numerous potentially irritating products. They are often associated with skin dryness. People with sensitive skin suffer from neurosensory symptoms such as feelings of heat, burning, tingling and itching, sometimes associated with redness. These skin disorders, coupled with disruptions to immune system and/or neurosensory mechanisms can be modulated or prevented by nutritional approaches and, in particular, through dietary use of certain strains of probiotics, including Lactobacillus paracasei NCC 246110,11.The mechanisms by which probiotics influence skin physiology are not yet entirely clear. As mentioned in reference to commensal bacteria, they may act on the immune system and induce a response in the host either by crossing the intestinal barrier, or by being captured, in the intestinal lumen, by immune cells that are able



SYNTHETIC BIOLOGY


SO

MM

AIR

E

to cross the gastrointestinal epithelium. The activated immune cells and cytokines released are thought to be transported to the skin, where they may modulate the inflammatory reactions caused by external physical and chemical irritants.

The underarm microbiome, axillary ecosystem and odours

The density of bacteria in the damp, occlusive and hairy area of the armpit is high, with an average of 2.5 million bacteria per cm2; the bacteria found there are mainly staphylococcus and corynebacteria (80% of the total bacterial population), anaerococcus and propionibacteria. For comparison, the density of bacteria on the forearm is only 4,500 per cm2.

Fresh sweat consists of a mixture of eccrine sweat (transpiration), apocrine sweat (pheromones) and a small amount of sebaceous secretion. Unpleasant smelling compounds are formed as a result of the enzymatic activities of bacteria and the influence of a number of other factors.

Understanding the axillary ecosystem of subjects whose underarm area is classed as unpleasant smelling or not, based on criteria that include the intensity of the odour, is necessary to identify imbalances in the components that make up this ecosystem. This understanding includes determining the biological signature or microbiome of armpits that produce an unpleasant odour and those that do not, or in other words, genetically identifying all the microorganisms (bacteria, yeasts and fungi) that proliferate in these ecosystems.

They differ in quantitative terms, with a higher level of bacteria present in armpits that smell unpleasant. These bacteria transform the fatty acids, amino acids and hormones present in the sweat released by apocrine glands into unpleasant-smelling compounds13.

The production of volatile compounds that have a smell varies between individuals and the disparity between odours and their intensity is based on numerous parameters. The first is gender: the precursors of volatile fatty acids and thiols are three times more prolific in men than women; secondly, age: adolescence is the period when the intensity of underarm odour is highest. This is due to the increase in the size and activity of the apocrine glands, accompanied by the hormonal changes that occur during puberty. Other factors are genetics, climate, dietary habits and hygiene.

Several strategies are used to reduce unpleasant odours. These include deodorants and antiperspirants, which change the microbiological signature of the armpit by destroying certain communities of bacteria. Masking or neutralising odours as they are produced is another approach. Grafting “good bacteria” is currently being examined to recreate an ecology in the underarm area that does not produce excessive body odour.

“The Smell of Us” project won an award at the final of the iGEM (International Genetically En-gineered Machines) synthetic biology competition for students held at MIT in 2014. The project presented by the Paris Interdisciplinary Research Centre outlined five different approaches to combating unpleasant body odours. Starting from the observation that it is not human sweat itself that smells unpleasant, but the substances produced when it is broken down by skin bacteria, the students worked on culturing “good” bacteria in the skin microbiome to mitigate or prevent the creation of unpleasant odours. L’Oréal researchers are most seriously involved in the approach called “Don’t sweat it”. It aims to change the microbiome by selecting naturally mutant skin bacteria, which could be incorpo-rated into a probiotic cream to reduce body odour.

The flora identified on the surface of

armpits with or without an unpleasant

smell are similar in qualitative terms.


1 Grice EA., Kong HH, Conlan S, Deming CB, Davis J, Young AC, Bouffard GG, Blakesley RW, Murray PR, Green ED, Turner ML, Segre JA. Topographical and temporal diversity of the human skin microbiome. Science 2009, 324 (5931), pp. 1190-1192).

2 Bouslimani A, et al. Molecular cartography of the human skin surface in 3D. PNAS, 30/03/2015.

3 Perez GI, Gao Z, Jourdain R, Ramirez J, Gany F, Clavaud C, et al. (2016) Body Site Is a More Determinant Factor than Human Population Diversity in the Healthy Skin Microbiome. PLoS ONE 11(4): e0151990. oi:10.1371/journal.pone.0151990

4 Ganju P, Nagpal S, Mohammed MH, Kumar PN, Pandey R, Natarajan VT, Mande SS, Gokhaleb RS. Microbial community profiling shows dysbiosis in the lesional skin of Vitiligo subjects. SciRep.2016;6: 18761. Published online 2016 Jan 13. doi: 10.1038/srep18761 PMCID: PMC4725359

5 Clavaud C, Jourdain R, Bar-Hen A, Tichit M, Bouchier C, Pouradier F, El Rawadi C, Guillot J, Ménard- Szczebara F, Breton L, Latgé JP, Mouyna I. Dandruff is associated with disequilibrium in the proportion of the major bacterial and fungal populations colonizing the scalp. PLoS One 2013;8(3):e58203.

6 Wang L, Clavaud C, Bar-Hen A, Cui M, Gao J, Liu Y, Liu C, Shibagaki N, Guéniche A, Jourdain R, Lan K, Zhang C, Altmeyer R, Breton L. Characterization of the major bacterial-fungal populations colonizing dandruff scalps in Shanghai, China, shows microbial disequilibrium. Exp Dermatol. 2015 ;24(5):398- 400

7 Soares RC, Camargo-Penna PH, de Moraes VC, De Vecchi R, Clavaud C, Breton L, Braz A1, Paulino LC Dysbiotic Bacterial and Fungal Communities Not Restricted to Clinically Affected Skin Sites in Dandruff. Cell Infect Microbiol. 2016 Nov 17;6:157. eCollection 2016.

8 Seite S, Flores GE, Henley JB, Martin R, Zelenkova H, Aguilar L, Fierer N. Microbiome of affected and unaffected skin of patients with atopic dermatitis before and after emollient treatment. J Drugs Dermatol. 2014 Nov;13(11):1365-72.

9 Gueniche, A., Knaudt, B., Schuck, E., Volz, T., Bastien, P., Martin, R., Röcken, M., Breton, L., Biedermann, T. Effects of nonpathogenic gram-negative bacterium Vitreoscilla filiformis lysate on atopic dermatitis: A prospective, randomized, double-blind, placebo-controlled clinical study (2008) British Journal of Dermatology, 159 (6), pp. 1357-1363

10 Guéniche A, Cathelineau AC, Bastien P, Esdaile J, Martin R, Queille Roussel C, Breton L. Vitreoscilla filiformis biomass improves seborrheic dermatitis. J Eur Acad Dermatol Venereol. 2008 Aug;22(8):1014-5. doi: 10.1111/j.1468-3083.2007.02508.x.

11 Gueniche A, Philippe D, Bastien P, Reuteler G, Blum S, Castiel-Higounenc I, Breton L, Benyacoub J. Randomised double-blind placebo-controlled study of the effect of Lactobacillus paracasei NCC 2461 on skin reactivity. Benef Microbes. 2014 Jun 1;5(2):137-45. doi: 10.3920/BM2013.0001

12 Benyacoub J, Bosco N, Blanchard C, Demont A, Philippe D, Castiel-Higounenc I, Guéniche A. Immune modulation property of Lactobacillus paracasei NCC2461 (ST11) strain and impact on skin defences. Benef Microbes. 2014 Jun 1;5(2):129-36. doi: 10.3920/BM2013.0014.

13 Callewaert C1, Kerckhof FM, Granitsiotis MS, Van Gele M, Van de Wiele T, Boon N. Characterization of Staphylococcus and Corynebacterium clusters in the human axillary region. PLoS One. 2013 Aug 12;8(8):e70538. doi: 10.1371/journal.pone.0070538


REFERENCES

t


SYNTHETIC BIOLOGY


SO

MM

AIR

E

tt

The objectives of synthetic biology are to optimise living material or even

produce it from scratch, using computers and microbial production

plants. As a result, applications in the fields of microbiome and exposome.

SYNTHETIC BIOLOGY

t

THE 21ST CENTURY REVOLUTION IN TECHNOLOGIES AND INNOVATION

Synthetic biology1, 40 years after the first genetic manipulations that allowed insulin to be produced by microorganisms, is no longer content with simply copying and pasting genes into bacterial cells. Its aim is to make new structures from fragments of DNA and assemble them like Lego bricks to create species that are not found in nature.

On the agenda are more effective treatments, less expensive drugs, renewable materials, biofuels, bacteria for pollution control, agriculture that can adapt to climate change and personalised foods for special diets.

All areas of activity have an interest in the field: the giants of the chemical, energy, agrobusiness and pharmaceutical industries are developing synthetic biology programmes alongside their counterparts in the IT sector, Microsoft and Google. The beauty industry, which is concerned about the environment and the sustainably produced raw materials, is also investing in the field, which makes it possible to produce high added-value materials such

as essential oils, fragrances and ingredients for cosmetics at a reduced cost.

Synthetic biology is taking advantage of both advances in molecular and cellular biology and increasingly efficient techniques for sequencing and manipulating DNA, while applying radically different methods from those used in biology until now, i.e. the techniques used by engineers, who view living organisms as a system and can therefore produce living machines, without being specialised in biology.

The possibilities for innovation offered by synthetic biology seem limitless and – similar to the situation that arose 40 years ago, with the advent of genetic engineering and genetically modified organisms – raise numerous safety, social and bioethical issues. Developing control systems (such as confining the new species created or generating a new genetic code that is incompatible with that found in nature) and appropriate regulations, as well as gaining public acceptance of the new field, are major challenges.

24L’Oréal 2017 - Research & Innovation / Synthetic Biology

Synthetic biology relies in large part on artificial synthesis of DNA

Synthetic biology programs or re-programs cells in the same way as programming a computer.The programme for all cells is contained in their genome (the DNA that contains all a cell’s genes). This tells them what they are for (their function) and what they have to do (produce a certain protein).

The fact that it is now possible to produce living material from scratch is due to the extraordinary progress made in sequencing technologies, which are now entirely automated. It is important to remember that DNA synthesis means first knowing its composition and using sequencing to define the number, type and order of the letters in the DNA (bases A, T, G and C) that make up genes. The efficiency of the techniques

used has doubled every year for the past ten years, with costs falling all the time. It took 13 years and €2 billion to sequence the human genome, which is made up of 20,000 genes and six billion ATGC bases. Today, an entire genome can be sequenced in a few weeks for a few thousand euros.

Giving the cell a new function means re-programming it by manipulating the genes that code the function. This in-volves creating virtual DNA sequences by using a computer program. The challenge of synthetic biology is isolating fragments of DNA, genes with specific functions, to make them into interchangeable parts like Lego bricks.

t

SYNTHETIC BIOLOGY: HOW DOES IT WORK?



SYNTHETIC BIOLOGY


SO

MM

AIR

E

Computer models

Mathematical models and computerised tools are needed not only to process the large volume of information created by sequencing operations but also to model and simulate the complex interactions between the components of living organisms, and design and predict the behaviour of biological systems before they are produced.

From the computer to biobricks

The information generated by sequencing, recorded in computerised databases and accessible on the internet, allows synthetic biologists to produce base units known as biobricks. Each of these standardised units has a specific function. Theoretically, it is possible to build anything you want from biobricks.

Biobrick synthesis is managed by a computer and the bricks are produced using a printer that extracts the various ATGC bases in the order indicated by the computer program. New bricks are created every year and made available to the entire community of synthetic biologists.

Assembling biobricks and inserting them into a frame to activate DNA

For DNA to be activated and become a living organism, DNA biobricks are joined together with enzymes, which act as biological activators. They are then inserted into a “biological frame” – a bacterium or yeast – so that they can function.

One of these frames is the bacterium Escherichia coli, which is a natural part of our intestinal flora. Like any procaryotic organism, it does not have a nucleus. This makes it easier to insert the new genetic material and read the new genetic programme carried in the biobricks.

In 2002, researchers at New York University recreated the genome of the polio virus from digitised sequences rather than from living material for the first time2. In 2010, following 15 years of research, the US biologist Craig Venter and his colleagues announced the creation of a self-replicating bacterial strain with DNA built entirely from computerised gene sequences.


Diagram of electronic analogs corresponding to biological components used in synthetic biology circuits. From https://lejournal.cnrs.fr/billets/labc-de-la-biologie-de-synthese

Synthetic DNA can be compared with software that is inserted into a computer (the frame). The genes and proteins that interact to fulfil cell functions are to the cell what electronic components (such as tran-sistors, condensers and resistors) are to the computer.

Components NetworksComputersModulesGates

Genes OrgansCellsPathwaysRéactions

t

PRODUCING A MOLECULE FROM A MICROORGANISM

27

Synthetic biology makes it possible to produce molecules using cellular production plants either by modifying existing living material, or by using a metabolic pathway that does not exist in nature.Both processes are illustrated below by the production of artemisinin, an antimalarial drug, and isobutene, a gas extracted from oil.

By reusing living material

Some chemical molecules can only be extracted from nature, for example from plants, fungi, insects or jellyfish and cannot be produced using synthetic chemistry methods. Synthetic biology provides a means of overcoming the difficulties or impossibilities associated with synthetic chemistry and reducing the number of steps required to

obtain a molecule using synthetic chemistry.

This is the case with artemisinin, extracted from the plant Artemisia annua (annual wormwood), which is mainly cultivated in Asia and used to treat malaria. Producing artemisinin using synthetic chemistry was dismissed because of the complexity of the process, which involved a dozen reactions, several of which are low in yield.

Jay Keasling, a professor at the University of Berkeley, modified a number of genes in baker’s yeast (Saccharomyces cerevisiae) by transferring genes from the plant that produced artemisinin into it. Following these changes, the yeast produced a chemical precursor of the drug3,4 using synthetic biology. The process was then industrialised. As a result between

L’Oréal 2017 - Research & Innovation / Synthetic Biology


SYNTHETIC BIOLOGY


SO

MM

AIR

E

60 and 100 tonnes of artemisinin have been produced annually in a production plant since 2013, representing between a third and a fifth of global needs.

By transforming biological systems that do not exist in nature

Today, it is no longer just genes that are transferred into a bacterium but biochemical circuits that connect proteins and DNA5. These can be metabolic circuits: these produce a chain of biological reactions that transform a natural component into a usable product, such as a drug, fuel or other product. They can also be regulatory circuits: in this case, they can trigger a particular type of production at the appropriate time, regulate it and improve its efficiency. Biochemical circuits can then be combined and integrated into tissues or living cells.

Isobutene, a gas that is currently extracted from oil, is used to produce plastics (PET used in bottles), rubber (for inner tubes) or fuel (diesel or kerosene).

Global Bioenergies, a company that grew out of the Génopole in Evry, has developed a process for producing isobutene from sugars of plant origin. Bacteria6 or yeasts are used to transform agricultural resources (such as sugar, cereals or agricultural wastes) into fuels and other materials. But since these microorganisms do not naturally produce hydrocarbons, they have to undergo major transformations to produce compounds that are identical to those derived from oil.

The synthesis process uses a metabolic pathway that does not exist in nature and has been introduced into a bacterium. In order to achieve this, researchers began by defining the chain of reactions that produces isobutene. Each chemical reaction triggers the action of an enzyme or biological catalyst. The code to produce each enzyme is contained in a specific gene identified by the researchers. The tool used for the design process was a computer. Thanks to this, the biologists were able to search databases for the DNA fragments that code enzymes, select them and carry out simulations.

After two years of research, the molecule was produced using fermentation. Isobutene produced in the form of a gas can be extracted continuously from the bacterial culture. This prevents any toxicity and makes it simpler to purify. Today, isobutene is being produced on an industrial scale.

In 2016, L’Oréal joined an industrial and commercial programme involving Global Bioenergies and the agro-industrial firm Cristal Union (a sugar-beet producer), focusing on the first biological production plant for isobutene. Derivatives of isobutene are widely used in the cosmetics sector.

Accessing sustainable bio-sourced products is one of the company’s environmental commitments. In fact, once it is used on a large scale, this innovative process will help to reduce carbon dioxide emissions into the atmosphere.

The applications of synthetic biology are considerable

In 2015, 116 products or industrial ap-plications based on synthetic biology were either already on the market or close to launch. These cover all biotechnological sectors, including drugs (artemisinin), diagnostic tools (AIDS and hepatitis), biofuels (from alga genes introduced into the bacterium E. Coli), biomaterials (isoprene used in manufacturing synthetic rubber and isobutene), pollution control (by deactivating certain genes that are not essential to their metabolism, Pseudomonas putida bacteria are forced only to consume and decompose toxic products) and detectors (E. Coli modified by synthetic biology to detect arsenic).

Industrial-scale production of the products derived from synthetic biology is a significant challenge, since it means obtaining the high yields compatible with commercial production.This means increasing the conversion yield of raw materials into the product of interest, improving productivity and optimising the product’s final concentration. The purification stages can represent up to 50% of the total production cost.


Some examples for the future...

Health: synthetic biology will be used to produce medicines that can currently only be obtained by extracting substances from plants, and will present reduced side effects.It will be used to develop personalised therapies with drugs that are adapted to each patient’s genome, treat metabolic imbalances (for example, excess urate, which circulates in the blood and is deposited in the joints, causing gout by establishing a biochemical circuit in the patient that converts the toxic product into a harmless substance), sensors for early detection of cancer cells, pathogenic agents that cause infections and regenerative medicine.

Energy: synthetic biology will be used to improve the fermentation of sugars by bacteria to produce ethanol, and improve microorganisms that can be used to synthesise hydrogen directly using photosynthesis.

The materials produced will be polymers and biodegradable plastics.

Agriculture: new plants that will be used as raw materials in the production of biofuels, plants that can adapt to climate change and plant protection treatments.


Genetically modifying bacteria so that they produce a cosmetic effect and making them into “improved probiotics” is an idea that is beginning to gain ground in the scientific community.

Reducing odours using synthetic biology

In the long term, current research could be used to develop products intended to control body odour by modifying the skin’s microbiome without a negative effect on beneficial bacteria. L’Oréal researchers from the Paris Interdisciplinary Research Centre collaborate with a team on “The Smell of Us” project presented in 2014. Starting from the observation that it is not human sweat itself that smells unpleasant, but the substances produced when it is broken down by skin bacteria, the students worked on culturing “good” bacteria to mitigate (or reduce) the creation of unpleasant odours by changing the skin’s microbiome.

Projects entered into the iGEM global student competition in synthetic biology (a competition created in 2005) in 2015, for example, propose modifying bacteria to target beauty problems.

A team from Technion (Israel) has developed a comb that dispenses two types of modified bacteria to combat androgenetic alopecia by breaking down dihydrotestosterone (a precursor of testosterone) directly at the hair root.

A team from the Californian hackerspace Biocurious, led by a former post-doc student from George Church’s laboratory, has taken its inspiration from the natural anti-UV protection observed in certain bacteria based on myscosporine-like compounds. They are working on a synthetic pathway that can be easily incorporated into any type of bacterium, for example to develop a filtering system as an alternative to “toxic” synthetic sun filters.

Chilean students are proposing a UV-sensor bracelet, which contains bacteria that produce a coloured compound when UV intensity is high, alerting the wearer that they need to protect themselves from the sun.Finally, a Canadian team has developed a bacterium that produces keratinase, mainly with the aim of helping to break down the human hair that blocks water treatment plants.

t

THE BEAUTY INDUSTRY AND SYNTHETIC BIOLOGY


SYNTHETIC BIOLOGY


SO

MM

AIR

E


1 D. E Cameron, C J. Bashor & J J. Collins. A brief history of synthetic biology. Nature Reviews Microbiology 12, 381–390

2 C J, Paul AV, Wimmer E. Chemical synthesis of poliovirus cDNA: generation of infectious virus in the absence of natural template. Science. 2002 Aug 9; 297(5583):1016-8.

3 C. J. Paddon et al., High-level semi-synthetic production of the potent antimalarial artemisinin. Nature 496, 528–532 (25 April 2013)

4 CJ. Paddon & JD. Keasling Semi-synthetic artemisinin: a model for the use of synthetic biology in pharmaceutical development. Nature Reviews Microbiology 12, 355–367 (2014)

5 N. Nandagopal, M B. Elowitz. Synthetic Biology: Integrated Gene Circuits. Science 02 Sep 2011: Vol. 333, Issue 6047, pp.1244-1248

6 B.N.M. van Leeuwen, A.M. van der Wulp, I. Duijnstee et al. Fermentative production of isobutene. Appl Microbiol Biotechnol (2012) 93: 1377. doi:10.1007/s00253-011-3853-7

REFERENCES


t


SYNTHETIC BIOLOGY


SO

MM

AIR

E

tt

Today, all over the world, the consumer can become an individual at the

heart of a system that combines science and technology, for whom

personalised cosmetics offer a response – in terms of products, skin and

hair care – which is suited to their needs and desires.

PERSONALISED COSMETICS

t

FROM OFF-THE-PEG TO MADE-TO-MEASURE

The research on skin and hair carried out at L’Oréal for over 100 years, the sequencing of the human genome, the discovery of the microbiome and revealing the importance of the impact of lifestyle on the individual’s health and well-being show that the established classifications used for criteria such as skin colour, hair texture, age and gender are inadequate when it comes to offering personalised cosmetic care.

Combining scientific knowledge with new data acquisition technologies, data-processing algorithms and industrial microproduction processes, to offer breakthrough products to every individual, are the issues in personalised cosmetics that L’Oréal is facing today.

From the consumer to the product designed specifically for them, the personalised cosmetics pathway consists of a series of steps, the complexity of which can be quite different depending on whether the product

is being offered following a simple online questionnaire or a biological or biochemical profile produced at a brand’s point of sale using a high-tech diagnostic tool.

L’Oréal has created a dedicated R&I unit to respond to the issues of personalised cosmetics, which operates as a technology incubator. The multidisciplinary team of around 15 people relies on L’Oréal’s 4,000 R&I researchers and collaborative ventures with start-ups all over the world to tackle every stage in the personalised cosmetics process.

Launching a personalised cosmetics product on the market is subject to the same regulations as any other cosmetics product, but with some much more significant constraints. In practice, the safety tests applied to every cosmetic formula also have to be applied to combinations of formulas. The dispensing mechanisms for personalised cosmetics are also subject to strict regulations.

32L’Oréal 2017 - Research & Innovation / Personalised Cosmetics


The consumer: Always at the heart of the system, the consumer is characterised by their biological identity: age, gender, phenotype (e.g. skin colour) and genotype. They live in a particular environment, in terms of geographical location, environmental pollution and UV index. They have a particular lifestyle, in terms of sleep, food and transport. Their specific physiological needs, expectations and changing desires are other factors that are taken into account in establishing their profile.

Analytical tools: These are used to produce an identity card for skin and hair. It could be a simple questionnaire, or a tool that analyses proteins in the epidermis, which combines science and technology.Originally developed for medical use, these tools have been adapted to meet the needs of the cosmetics sector. For example, a reader that measures glycaemia has been adapted to analyse biological markers, DNA or proteins in the hair or skin.

Miniaturised items of laboratory equipment often become connected portable objects, such as scanners, biosensors, imaging and biometric analysis devices based on recognising characteristics that are specific to the individual. The measurements obtained can be stored and analysed directly using dedicated software.

As the technology develops, these analytical tools are providing increasingly accurate, reliable and fast measurements. Analysing physical parameters provides information on the apparent or underlying condition of skin or hair. Skin colour, wrinkles, redness, elasticity and areas of pigmentation, and hair colour, shine and surface condition are assessed using imaging tools and biosensors that provide objective information to complement clinical assessments.

Data processing algorithms: Algorithms are series of calculations that incorporate all the data available: biological and physical measurements carried out using analytical tools, environmental data, and in some cases consumer preferences and the active ingredients used in product formulas. Ultimately, they provide a personalised or individualised product solution.Complex algorithms of this kind offer considerable added value in personalising cosmetics and patents have been filed to protect them.

They are used to find or create a colour which, when applied to the hair or skin, will reproduce the exact shade required in daylight; they might link a biological marker, for example a protein in the epidermis, to a skin ageing symptom, and propose the right level of active ingredients to reduce the sign of ageing; they might also link the value of a UVA and UVB measurement to the level of sun protection recommended.

Dispensing personalised care: The product or treatment designed to suit the consumer’s profile will be made by mixing pre-existing formulas either manually or using an automated process. Automation means designing high-tech dispensers. Personalising formulas that are produced individually requires very accurate measuring. Because pack volumes are small (10 to 100 ml), the quantity to be added is the total of some tiny amounts of certain compounds. These are previously mixed into bases that are then added to the product to overcome this difficulty and reduce the number of ingredients (preservatives, etc.).

Delivering advice on the amount of product to be used: for the UV patch, for example, a smartphone app indicates the amount of UV protection required to provide active protection from UV radiation.

THE FIVE CRITERIA FOR PERSONALISED COSMETICS


SYNTHETIC BIOLOGY


SO

MM

AIR

E

The five examples of personalised cosmetics below take us into a

connected, high-tech world, where the scientific knowledge of skin and

hair developed for decades in L’Oréal’s research laboratories is used to

establish the consumer’s personal profile.

t

FIVE EXAMPLES OF PERSONALISED COSMETICS

A foundation that matches their skin colour.

Personalised skin care adapted to the skin’s physiology in its environment

My UV Patch: a flexible sensor that is stuck to the skin to measure exposure to UVA and UVB.

Makeup Genius : a transformative tool that of-fers virtual testing of make-up products by vis-ualising the result in advance, with an extremely realistic colour rendering.

A smart connected hair brush

Personalised foundation Le Teint Particulier by Lancôme

A beauty adviser at the point of sale measures skin colour precisely using a scanner and defines the consumer’s preferences. The data are processed using an algorithm – which indicates the make-up shade that is best suited to the skin colour – and instruct a machine to produce the correct combination. Peristaltic pumps dispense the right combination of base and pigments into an airtight bottle. The client leaves with her personalised foundation, stamped with her reference number for her next refill.

Consumer studies show that women are not satisfied with the foundation colours that are predetermined by each manufacturer and want a colour that matches their skin tone exactly.

Formulating the most appropriate shades of foundation means it is essential to know exactly what skin colours there are. A study carried out by L’Oréal quantified skin colours around the world by measuring the skin tone of over 2,000 women in 10 different countries, and then cat-egorised them independently of any statement about belonging to a particular ethnic group1. Based on these results, it is possible to pro-duce shades that are increasingly close to skin colours and create a truly “nude” look.

The profiling tool used by the beauty adviser at the point of sale is a scanner that measures skin colour precisely at several points on the face: forehead, cheeks and jaw.

Measuring skin colour involves determining three parameters: shade (from red to yellow on the spectrum), brightness (from light to dark) and saturation (intensity)2.

A colour equation algorithm analyses the meas-urements to find the perfect shade of make-up for a particular skin colour. The data processed for the formulation are sent via blue tooth to a ma-chine that produces a personalised liquid foun-dation at the point of sale in just a few minutes.


25 30 35 40 45 50 55 60 65

20 000 -

30 000 -

40 000 -

50 000 -

60 000 -

70 000 -

80 000 -

CAUCASIANWOMAN

HISPANICWOMAN

ASIAN WOMAN

AFRICANWOMEN

INDIANWOMAN

Lightness

Dark

Light

Hue

Yellow Red

A beauty adviser produces an identity card for the consumer’s skin at the point of sale, following a comprehensive analysis of the skin surface carried out using connected portable tools, such as scanners or biological marker analysers. Algorithms analyse the data obtained and incorporate additional data on the environment and the consumer’s preferences. The result of processing and cross-referencing the data is a care formula that is suitable for the skin’s physiology in its specific environment, produced by a machine at the point of sale. The client leaves with her personalised skin care product, stamped with her reference number for her next purchase.

Personalised skin care


The machine does not replicate the entire com-plex process of formulation when it produces a foundation. The system mixes three basic pre-existing foundation colours and dispenses the resulting colour into a special type of pack-aging. The colour is an exact match for the one scanned. In order to avoid oxidation and en-sure colours stay accurate, the Le Teint Partic-ulier device has eight peristaltic pumps, which prevent air getting into the mix of formulas and dispense it. The formula can be enhanced by adding a moisturiser and a mattifier for oily skin. Skin colour chart2

Each individual’s skin is unique. Firmness, skin hydration and consistency of colour are some of the skin characteristics that vary not only with age but also the environment, seasons

and lifestyle. The type of care needs to be suit-able for the skin’s physiology in its environment. It also needs to align with the consumer’s own preferences.


SYNTHETIC BIOLOGY


SO

MM

AIR

E

L’Oréal 2017 - Research & Innovation / Personalised Cosmetics36

L’Oréal 2017 - Research & Innovation / Personalised Cosmetics

My UV Patch: the flexible UV sensor

The first flexible UV sensor designed to help consumers use the right kind of sun protection

for their skin type and exposure to UV radiation. This ultra-thin, self-adhesive patch includes

photosensitive colouring agents that change colour depending on the level of UV they receive

and the colour of the skin it is placed on. It measures instant and cumulative levels of UV over

several hours or for up to a few days, and tells the wearer if their UV protection is inadequate.

Every individual has their own particular phe-notype, which includes their skin colour. This determines the level of sun protection needed during exposure to UV radiation. Understand-ing the level of UV received daily or over a pe-riod, and the type of photoprotection needed, are part of consumers’ expectations. They also expect not to be able to feel a patch when it’s in contact with their skin and to be able to wear it in all circumstances, i.e. it needs to be resistant to water, sweat and sun cream, which can be applied directly on top of it.Unlike the rigid technologies currently availa-ble on the market, My UV Patch is a flexible,

transparent film that adheres to the skin. With a surface area of approximately 2.5 cm² and 50 micrometres thick, the heart-shaped patch, made up of squares, includes photosensitive colouring agents hat form a pattern and act as dosimeters: the blue in the colouring agents changes according to the level of UV received and the colour of the skin the patch is placed on. The colouring agents currently available on the market reach saturation point very quickly (they change colour after 20 minutes in sun-light, so the patches are no longer usable). As a result, L’Oréal chemists have developed spe-cific (patented) formulas for the eight different

37

The condition of facial skin is analysed using portable connected tools. Scanners that are still only accessible to professionals at the point of sale or already available for personal use by the consumer assess the skin’s surface. Thanks to non-invasive analytical methods us-ing light, and taking pictures of several areas of the face, scanners can accurately determine the colour of the skin, identify areas of redness, assess the depth of wrinkles and the cumu-lative damage caused by sun exposure over time, and detect the presence of bacterial flora.

Other devices measure biological markers, for example quantifying one or more proteins in the epidermis based on a non-invasive sam-pling of facial skin. These proteins reflect the level of skin hydration, firmness or oxidative stress. The consumer is given an identity card for their particular skin.

Algorithms are used to process the data and incorporate additional information about the environment, hygrometry, pollution, UV index and the level of active ingredients in the associ-ated pre-existing formulas.

The product best suited to the consumer’s needs, based on the analyses produced using portable tools and their preferences, is made at the point of sale using a machine that com-bines different bases containing antioxidants, moisturisers and active ingredients to increase the density of the dermis.

The consumer can also personalise her own skin care by adding boosters (such as antioxi-dants and moisturisers) to her products, which are offered on an ‘à la carte’ basis by some cosmetics brands.


SYNTHETIC BIOLOGY


SO

MM

AIR

E

Makeup Genius An augmented reality make-up simulator for personalised use

Makeup Genius is a make-up app that uses facial tracking technology to transform the screen on a smartphone or tablet into a virtual mirror. The user’s application of virtual make-up is rendered in real time using the smartphone’s front-facing camera; the user can see the result of the make-up on their face and buy the product they have tested directly online if they wish.

colouring agents, which can change colour at different intensities. One changes colour af-ter 20 minutes; the others change after eight hours, a day, three days and five days. Eight other squares act as reference colours for cali-brating the light. Moreover, all the other self-adhesive patches on the market rely on a visual warning. Thanks to a specific (patented) algorithm, My UV Patch interacts with a smartphone to quantify UVA and UVB values. An NFC (Near Field Communication) chip ac-tivates the dedicated “Love your skin” app on the user’s smartphone. By scanning their patch, the user has access to accurate data

on their degree of exposure to all types of UV radiation (UVB, UVA and long UVA) and is given advice. The app tells the consumer when they will have to reapply their sun screen according to dermatological recommendations.

The patch is the result of a partnership be-tween L’Oréal’s Connected Beauty incubator and the company MC10, Inc for the patch,, and the start-up PCH for the industrial scale-up. L’Oréal has contributed its expertise in al-gorithms and its knowledge of the cosmetics market.

It was launched under the dermatological brand La Roche-Posay.


There is a significant gap between women’s expectations of how make-up is presented and the results they get when the product they have bought is tested in real conditions. More-over, although sales outlets offer a large num-ber of products, they may not sell the full range and the product the consumer wants may not be available. The Makeup Genius app allows users to scan a L’Oréal Paris product to find a colour that suits them, try it virtually and pur-chase it online. It also allows them to try out the looks created by L’Oréal Paris professional make-up artists and share them on social me-dia.

Based on complex algorithms and a facial rec-ognition system, Makeup Genius captures 64 points on the face so that it is possible to dis-tinguish lip skin from the skin around the eyes or on any other part of the face. This allows the virtual make-up applied to move with the face, change your expression and test different looks from different angles and under different lighting conditions.

The research team tested 4,000 lighting con-ditions and collected 180,000 images taken using the app on several types of camera to “be sure that what you see is what you’re going to get” on all skin tones – based on five ethnic groups with skin colours ranging from very pale to very dark – in natural and artificial lighting conditions and from every angle.

L’Oréal researchers have worked in collabora-tion with a team of scientists from Image Met-rics – whose facial mapping technology was used in the film The Curious Case of Benjamin Button – to incorporate the facial tracking algo-rithm into the app. This was a first, since until then, the technology had only been used in Hollywood and the 3D video games industry.

The Makeup Genius app launched in 2015 was followed by other versions, for foundation (Shade Genius) and nail varnish (Nail Genius) in 2016.

A study published by scientists at L’Oréal showed that brushing too vigorously damages the hair, makes the fibre brittle and causes split ends. The connected brush minimizes these risks, by means of a series of sensors that col-lect data about the quality of the hair and how best it should be brushed.

A microphone records the sound generated by brushing, and captures models to provide information about the appearance of frizz, split ends and breaks. It also provides data about handling, suppleness or dryness.

Strength sensors measure the force applied to the hair and scalp during brushing.

An accelerometer and a gyroscope analyse in detail the brushing method and count the number of brush strokes. The user is alerted when these movements become too harsh.

Connectivity sensors analyse the environment and determine whether the hair is dry or damp.

Just brushing is enough to start the data col-lection process. The sensors transmit the data collected via Wi-Fi or Bluetooth automatically to a special mobile application. Weather con-ditions such as humidity, temperature, UV and wind that influence hair styling and the hair are included in the data.

A smart connected hair brush A technology that transform the hair beauty routine

Kerastase Hair Coach Powered by Withings, the first connected brush, evaluates the quality of

the hair and measures the effects of various hair-care routines by means of sensors and signal

analysis algorithms. It comes with a mobile application used to access additional data and

custom product recommendations in order to improve hair health.



SYNTHETIC BIOLOGY


SO

MM

AIR

E

40

The bathroom of the future will be connected

The mirror will be the essential feature in the bathroom of the future. It will be a connected

object, made up of LCD screens linked to a computer, and will act as a 3D scanner to examine

the person standing in from of it from a physiological and physical point of view, including their

lifestyle and in a particular environment. It will offer beauty advice, store the data it gathers

and be able to programme the machine to produce personalised care or place an order at the

chosen point of sale.

The morning weather forecast will provide in-formation about the day’s temperature, level of pollution and UV index and recommend the right level of sun protection needed.The user’s UV patch can provide this informa-tion on request all day long.

The scanner will measure skin colour, detect areas of pigmentation, redness, the depth of wrinkles and the degree of skin hydration... and send the information, which will be stored and used by the mixing device to create a day cream combining antioxidants, pro-collagen active ingredients, etc.

An app will provide advice on the right make-up to use for the day, based on the clothing and hairstyle the user has chosen.

A set of connected scales will provide data on body mass, store them for a monthly state-ment, and provide dietary advice.

A connected bracelet will send data about the user’s physical activity, sleep time, cardiac rhythm and stress level, and in return recom-mend physical activities for the day.

A connected brush will provide information on the condition of the user’s hair, read its colour, identify white or bleached hair and suggest shades for next time it is coloured or personal-ised care to repair hair that has been damaged by the sun.

The mobile application analyses how users brush their hair and takes account of their habits and the environment to provide data on the hair quality index, the efficacy of the hair brushing gesture and personalized advice and recommendations on Kérastase products and hair care.

Developed for Kérastase, L’Oréal’s expert hair-care brand, the connected brush is the fruit of a partnership between Withings, a French start-up company for the sensors used in the brush and L’Oréal’s research Tech Incubator for the signal analysis algorithms. It will be available from the middle of 2017.

L’Oréal 2017 - Research & Innovation / Personalised Cosmetics41



SYNTHETIC BIOLOGY


SO

MM

AIR

E

42

1 Caisey L, Grangeat F, Lemasson A, Talabot J, Voirin A. Skin color and makeup strategies of women from different ethnic groups. Int J Cosmet Sci. 2006 Dec;28(6):427-37. doi: 10.1111/j.1467- 2494.2006.00329.x.

2 de Rigal J, Abella ML, Giron F, Caisey L, Lefebvre MA. Development and validation of a new Skin Color Chart. Skin Res Technol. 2007 Feb;13(1):101-9.

3 Webb RC, Pielak R2, Bastien P3, Ayers J, Niittynen J, Kurniawan J, Manco M, Lin A, Cho NH, Malyrchuk V, Balooch G, Rogers JA. Thermal transport characteristics of human skin measured in vivo using ultrathin conformal arrays of thermal sensors and actuators. PLoS One. 2015 Feb 6;10(2):e0118131. doi: 10.1371/journal.pone.0118131. eCollection 2015

4 Webb RC, Bonifas AP, Behnaz A, Zhang Y, Yu KJ, Cheng H, Shi M, Bian Z, Liu Z, Kim YS, Yeo WH, Park JS, Song J, Li Y, Huang Y, Gorbach AM, Rogers JA. Ultrathin conformal devices for precise and continuous thermal characterization of human skin. Nat Mater. 2013 Oct;12(10):938-44. doi: 10.1038/nmat3755. Epub 2013 Sep 15.

5 Luengo G, Galliano A, Dubief C. (2014), Aqueous Lubrication in Cosmetics. Hackensack, New Jersey : World Scientific Publishing

REFERENCES

t


Author Catherine GERSTArt Direction Isabelle WALTERIllustrations Fiamma LUZZATI

Graphic Design WalkandCreate.com© L’Oréal Research & Innovation 2017


SYNTHETIC BIOLOGY


SO

MM

AIR

E

Scientific Directoratewww.loreal.com

http://www.loreal.com

science, technology & beauty 2017 -...

Documents