Francis Quinn

L’Oréal Research

11 April 2008

NANOTECHNOLOGY IN COSMETICS

NANOTECHNOLOGY

Natural nanoparticles Synthetic nanoparticles

WHAT ARE THE NANOTECHNOLOGIES USED IN COSMETICS ?

The types of products developed on the nanometric scale by the cosmetic industry are different from nano materials used in other industries.

They differ by their shape and their molecular structure, their mode of use and their specific interactions with the living world and the environment.

In essence, they are nanoemulsions and nanopigments.

WHAT ARE NANOEMULSIONS ?

Nanoemulsions are found widespread in nature, such as milk. In cosmetics, they are in fact macroscopic preparations containing oil and water droplets reduced to nanometric size to increase the content of nutritious oils while preserving the transparency and the lightness of the formulas.

Sometimes fragile active ingredients, like vitamins, are protected from air inside vesicles called nanocapsules™ or liposomes that release the ingredient upon contact with the skin at the time ofapplication.

Nanoemulsions therefore do not cross the skin barrier1-3. It is widely acknowledged that nanoemulsions, that are liquid particles and not solid objects, do not expose users to health risks4.

1) Imbert D and Wickett R: Topical delivery with liposomes. Cosmetics and Toiletries magazine. 1995; 111:32-45.

2) Honeywell-Nguyen P et al.: Quantitative assessment of the transport of elastic and rigid vesicle components and a model drug from these vesicle formulations into human skin in vivo. Journal of Investigative Dermatology. 2004; 123(5):902-10.

3) Van den Bergh B et al.: Interactions of elastic and rigid vesicles with human skin in vitro: electron microscopy and two-photon excitation microscopy. Biochimica and Biophysica Acta. 1999; 1461:155-173.

4) United States Food and Drug Administration, Nanotechnology Report 2007 (see www.fda.gov/nanotechnology/taskforce/report2007.pdf); British Standards Institute, PAS 130:2007, Guidance on the labelling of manufactured nanoparticles and products containing manufactured nanoparticles.

Polymer

NANOEMULSIONS, LIPOSOMES & NANOCAPSULES

Nanocapsule™150-600 nm

(Vitamin A,E)

Nanoemulsion50-80 nm

(Nourishing oils, unique texture & transparency)

Water

Lipid

Liposome150-300 nm(Caffeine)

Water

Oil

TITANIUM DIOXIDE NANOPIGMENTS

Nanopigments such as titanium dioxide (TiO2) are minerals already present in our natural environment, in the form of clay for example.

Titanium dioxide is one of the best known. It is a totally insoluble, inert material. It is a reference of non toxicity which is why we find it used widely in foodstuffs (colouring agent E171) as well as dental hygiene products including toothpaste.

Titanium dioxide used in sunscreens because it is renowned for its capacity to neutralise UV radiation.

SUNSCREENS & SKIN CANCER PREVENTION

Many studies show that the use of sunscreens reduces the occurrence and the development of skin and lips cancers, and herpes labial1-3.1) Pogoda J, et al.: Solar radiation, lip protection, and lip cancer risk in Los Angeles County women (California, United States). Cancer Causes Control 1996, 7(4): 458-463.2) Rooney J, al.: Prevention of ultraviolet-light-induced herpes labialis by sunscreen. Lancet 1991, 338(8780): 1419-1422.3) Nohynek G, Schaefer H.: Benefit and risk of organic ultraviolet filters. Regul. Toxicol. Pharmacol. 2001, 33: 285-291.

Sunscreens protect DNA from deteriorations induced by ultra-violet radiations, reduce the appearance of modified squamous cells and the development of certain indices of the melanoma4-8.4) Young A, et al.: Protection by Ultraviolet A and B sunscreens against in situ dipyrimidine photolesions in human epidermis is comparable to protection against sunburn. J. Invest. Dermatol. 2000, 115: 37-41.5) Gallagher R, et al.: Broad-spectrum sunscreen use and the development of new nevi in white children. A randomized controlled trial. JAMA 2000, 283: 2955-60.6) Gallagher R.: Sunscreens in melanoma and skin cancer prevention. CMAJ 2005, 173(3): 244-245.7) Mahroos M, et al.: Effect of sunscreen application on UV-induced thymine dimmers. Arch. Dermatol. 2002, 138(11): 1480-1485.8) Lee T, et al.: Site-specific protective effect of broad-spectrum sunscreen on nevus development among white schoolchildren in a randomized trial. J. Am. Acad. Dermatol. 2005, 52: 786-792.

SUNSCREENS & SKIN CANCER PREVENTION

Today, 1 cancer in 3 is skin cancer.

Each year 2 to 3 million skin cancers (carcinoma type) and over 130,000 melanomas are diagnosed.

For example, 1 in 5 Americans will develop a skin cancer in his life.

The incidence of skin cancer has increased considerably over the last decade. The popularity of outdoor activities, including “sun bathing” is the leading cause of this increase.

The reduction of the level of protection from the ozone layer, if continued, will worsen this tendency: it is estimated that a decrease of 10% of stratospheric ozone would be responsible for 300,000 carcinoma and 4,500 additional melanomas each year. See World Health Organisation at http://www.who.int/uv/health/uv_health2/en/index1.html

SUNSCREENS & SKIN CANCER PREVENTION

In many countries dermatologists' associations and national sanitary authorities regularly launch information campaigns inviting people to apply sun products before exposing themselves to the sun.See The Cancer Council Australia at http://www.cancer.org.au/content.cfm?randid=906824

For example, in 2006 this public health strategy was re-confirmed by The American Academy of Dermatology. See the American Academy of Dermatology at http://www.cancer.org/docroot/PED/content/ped_7_1_Skin_Cancer_Detection_What_You_Can_Do.asp

According to the IARC (International Agency for Research on Cancer), the regular use of sun creams is essential in the prevention of the skin cancers and those that contain nano titanium dioxide or zinc oxide are the most effective.See IARC Handbooks of Cancer Prevention: Sunscreens. International Agency for Research on Cancer, World Health Organization, Lyon 2001.

NANOSIZE ENHANCES UV SCATTERING / REFLECTION OFTITANIUM DIOXIDE DISPERSIONS

All products diluted in Cyclopentasiloxane (0.001% TiO2). Spectrophotometer Hitachi U-3310 - Integrating Sphere.Delrieu P, et al.: Perspectives on Supplying Attenuation Grades of Titanium Dioxide and Zinc Oxide for Sunscreen Applications FDA Public Meeting on Nanotechnology, 10 October 2006, Washington DC.Data made available by KOBO, www.koboproducts.com

CONSUMER USE & RE-USE

The increased transparency of sunscreen products containing nanopigments facilitates their acceptance and (re-)application by consumers, and consequently improves protection against UVA and UVB for all skin types.See IARC Handbooks of Cancer Prevention: Sunscreens. International Agency for Research on Cancer, World Health Organization, Lyon 2001.

195 60 35 15 10 nm

195 60 35 15 10 nm

195 60 35 15 10 nm

195 60 35 15 10 nm

The titanium dioxide nanopigments used in sunscreen invariably form clumps of size much greater than 100 nm.

A large number of studies have demonstrated the titanium dioxide nanopigments do not cross the skin barrier after topical application1-9.

Studies, including those carried out within the framework of the European research program NANODERM, have clearly shown that nanopigments do not cross the skin barrier of either healthy skin or compromised skin (acne, psoriasis)10-14.

PROTECTING THE SKIN SURFACE

STUDIES SHOWING THAT TITANIUM DIOXIDE NANOPIGMENTS DO NOT CROSS THE SKIN BARRIER

1) Lademann J, et al.: Penetration of titanium dioxide microparticles in a sunscreen formulation into the horny layer and the follicular orifice. Skin Pharmacol. Appl. Skin Pyysiol. 1999, 12: 247-256.

2) Dussert A, et al.: Characterization of the mineral content of a physical sunscreen and its distribution onto human stratum corneum. Int. J. Cosm. Sci.. 1997, 19: 119-129

3) Pflucker F, et al.: The outermost stratum corneum layer is an effective barrier against dermal uptake of topically applied micronized titanium dioxide. Int. J. Cosm. Sci. 1999, 21: 399-411.

4) Gamer A, et al.: The in vitro absorption of microfine zinc oxide and titanium dioxide through porcine skin. Toxicology in Vitro. 20, 301-307, 2006.

5) Roberts M.: Nanoparticles In Topical Products – A Consumer Health Risk? FDA Public Meeting on Nanotechnology, 10 October 2006, Washington DC.

6) Nohynek G, et al.: Grey goo on the skin? Nanotechnology, cosmetic and Sunscreen Safety. Crit. Rev. Tox. 2007, 37:1-27.7) Mavon A, et al.: In vitro Percutaneous Absorption and in vivo Stratum Corneum Distribution of an Organic and a Mineral

Sunscreen. Skin Pharmacol. Physiol. 2007, 20:10-20.8) Stern S, et al: Nanotechnology safety concerns revisited. Tox. Sci. 2008, 101:4-21.9) N. Sadrieh N, et al.: Analysis of Dermal Penetration of Titanium Dioxide (TiO2) from Sunscreen Formulations Containing

Micron- and Nano-scale Particles of TiO2. Presented at the US Society of Toxicology meeting, Abstract n° 1023, Charlotte, 2008.

10) Butz T, et al.: No evidence for nanoparticle penetration into living skin. Preliminary data presented at ECETOC, Barcelona, 2005.

11) Pinheiro T, et al.: The influence of corneocyte structure on the interpretation of permeation profiles of nanoparticles across skin. Nucl. Instru. and Meth. in Phy. Res. B 2007, 260:119–123.

12) Borbala K, et al.: Investigation of micronized titanium dioxide penetration in human skin xenografts and its effect on cellular functions of human skin-derived cells. Exp. Derm. 2008, DOI:10.1111/j.1600-0625.2007.00683.x

13) Filipe P, et al: Nanotoxicity of TiO2 and ZnO containing sunscreens versus the stratum corneum barrier dogma. Expt. Tox. 2008 in press

14) Sugibayashi K, et al.: Safety evaluation of titanium dioxide nanoparticles by their absorption and elimination profiles. J. Tox. Sci. 2008, in press

TITANIUM DIOXIDE NANOPIGMENTS

Nano titanium dioxide clumps

Nano titanium dioxide clumps from a sun cream on the skin surface

SIZE DISTRIBUTION OF “15 nm”TITANIUM DIOXIDE NANOPIGMENT

500 nm

More importantly, a recent study carried out in the US by the FDA demonstrated that in the case where titanium dioxide nanopigments are injected subcutaneously or even directly into the blood stream, no adverse effects no signs of toxicity, no inflammation. Furthermore, no nanopigments are found in the brain. The nanopigments are eliminated normally by the phagocytic barrier1.

These results were confirmed by an independent European study2.

In fine, the question as to whether nanopigments could possiblycross the skin barrier may not be so central…

Indeed, if nanopigments do not induce unwanted effects when theyare injected intravenously, how could pose a health risk when there are simply put in contact with skin?

RECENT STUDIES BY THE US FOOD AND DRUG ADMINISTRATION (FDA)

1. Umbreit T, et al.: toxicology of titanium dioxide nanoparticles: 1 Characterization and tissue distribution in subcutaneously and intravenously injected mice. Presented at the US Society of Toxicology meeting, Abstract n° 1386, Charlotte, 2007.

2. Fabian E, et al: Tissue distribution and toxicity of intravenously administered titanium dioxide nanoparticles in rats. Arch. Toxicol. DOI 10.1007/s00204-007-0253-y

Applying the precautionary principle, L’Oréal does not market products containing either nanotubes or fullerenes.

Why? Because the available scientific studies cannot guarantee no unwanted side-effects. Furthermore, the relation cost / efficiency is questionable.

Furthermore, L’Oréal does not put nanomaterials into its aerosol products, because of secondary effects described in the scientific literature after inhalation of some nanoparticles.

L’OREAL AND THE PRECAUTIONARY PRINCIPLE

Clearly any new technology has to be developed and applied taking all the necessary precautions to control the potential risks.

However, we also have to allow the researchers to optimize the benefits of scientific innovation in interest of the public and the environment.

It is therefore vital to put into place an international framework, that includes civil society groups, to establish standards in the area of nanotechnology.

Furthermore, the standards should be pertinent and adaptedaccording to the various industries that use and apply the technology.

Through ICON and Afnor, L’Oréal is an committed partner is this process that will take some time to complete fully.

INTERNATIONAL STANDARDS AND NORMS

The absence of specific standards for nanotechnology is regrettable because the public without expert knowledge cannot distinguish between the different types of nanomaterials used by different industries for which the uses ― and the potential risks ― are very different.

This lack of transparency in the nomenclature of nanos leads to doubts, rumours, even fantasies, about both the benefits and therisks.

Furthermore, the excessive and sometimes deceitful use of the term "nano" penalises the real innovations coming from serious private companies and public laboratories.

WHY ARE STANDARDS IMPORTANT ?

L’Oréal has been using nanotechnology in its products safely for over 25 years ― nano titanium dioxide in sun creams and nanocapsules were first commercialized in early 1980’s.

L’Oréal applies the precautionary principle and believes that while any new technology must be developed and applied taking all the precautions necessary to minimize the possible risks, one must also allow researchers to optimize the benefit of scientific innovation in the interest of the public and the environment.

One cannot overstate the need and the importance of an international consensus that includes civil society participation for the development of standards for nanotechnologies that are relevant and adapted to the various industrial sectors.

CONCLUDING REMARKS