Recent updates in cosmetic technology were discussed. Key points include: - Safety is the top priority in cosmetic formulation given skin's sensitivity. Animal testing is being phased out and alternatives like in vitro tests are being validated. - Nanotechnology shows promise for cosmetics through nanoemulsions, liposomes and other nanoparticles that can improve delivery and stability of active ingredients. Potential risks need consideration. - Stem cell cosmetics are an emerging area but current products do not contain actual stem cells. Regulations are still maturing around cell manipulation and product characterization for these types of cosmetics.

1. Recent Updates in Cosmetic
Technology
Dr. Khaled Al-Qaoud
R&D manager
Jordan Company For Antibody Production
(MONOJO)

2. Personal Care
• ((For the first time, the world largest personal
care market serving 500 million consumers
will be served by a single, harmonized piece of
legislations, directly applicable in all member
states of the EU))
• Bertil heerink; director general of cosmetic
Europe, July 2013.

3. Definition – Cosmetics
• Articles intended for:
– Cleansing
– Beautifying
– Promoting attractiveness
– Altering appearance

4. What are quantitative Ways for Its
Effect
• Protection against sun radiation
• Moisturization/hydration of the skin
• Reduction of skin roughness
• Reflection of light from the skin surface
• Water loss changes
• Effect on skin
– temperature
– pH
– Topography
– Roughness
– Friction
• Analysis of the cell layers of the stratum corneum and
spinosum

5. Aging is a matter of skin Protein
deformation
Elastin and
Collagen
proteins are
main players

6. Major Concerns in Cosmetic Field
• Irritation
• Allergic Reaction
• Mutagenicity, cancer development

7. Safety First
• Cosmetics are principally concerned with
FORM rather than FUNCTION.
• FORMULATION is a challenge
• Every COMPONENT should complies with the
relevant regulations.
• Deep understanding of problems linked to
release of active compounds on cutaneous
level.
• What if SKIN is problematic

8. Animal Models For Skin Irritation and
Topical Toxicity
• Guinea Pig Maximization Test (GPMT), The OECD
Guidelines for the Testing of Chemicals guideline No. 406 of 1992
• Local Lymph Node Assay (LLNA), OECD Guidelines for the
Testing of Chemicals guideline No. 429 of 23 July 2010
– first-choice method for in vivo testing
• Draize Rabbit Eye Test for predicting the ocular
irritation/toxicity
• Mouse Ear Swelling Test

9. Skin Irritation Studies

10. MONJO Rabbit Ear Irritation Model

11. European Ban on Animal-Tested Cosmetics
Finally Set for Implementation in March, 2013

12. What is banned in Europe related to
animal use
• Testing of cosmetics on animals
• sale of animal-tested cosmetics throughout the
EU from 2009
• cosmetics-related animal testing
• Imported cosmetics ingredients tested on animals
(2013)
2013, India announced a ban on testing cosmetics
on animals
2014, Brazil followed

13. Animal Testing & Cosmetics
FDA
• FDA is responsible for assuring that cosmetics are
safe and properly labeled (Federal Food, Drug,
and Cosmetic Act (FD&C Act))
• does not specifically require the use of animals
in testing cosmetics for safety
• does not subject cosmetics to FDA premarket
approval
• Agency advise cosmetic manufacturers to employ
whatever testing is appropriate and effective for
substantiating the safety of their products
Including animal use

14. Animal Testing & Cosmetics
• FDA supports and adheres to the provisions of
applicable laws, regulations, and policies governing
animal testing, including the Animal Welfare Act and
the Public Health Service Policy of Humane Care and
Use of Laboratory Animals
• Maximum useful scientific information from the
minimum number of animals
• consideration should be given to the use of
scientifically valid alternative methods to whole-
animal testing
• In 1997, FDA joined with thirteen other Federal
agencies in forming the Interagency Coordinating
Committee on the Validation of Alternative Methods
(ICCVAM).

15. In vitro tests for skin sensitization
• Keratinocyte cultures
• Langerhans cell cultures
• Peripheral blood derived dendritic cells
• Cell lines

16. Alternatives Non-Human-Animal
testing
• Human 3D skin models
• Monitoring DNA, RNA changes
• Interleukins determination
• Mutagenic testing on cell lines

17. Genotoxicity and Mutagenic Cosmetics
Chromosomal
damage due
to toxic agent

18. Stain to visualize

19. Skin Hypersensitization

20. Cascade reaction for skin sensitization

21. Allergen
study on
human
skin

22. Skin Hypersensitization molecule level
good correlation
between
activation of the
Nrf2 signaling
pathway and
chemical
sensitization was
first shown by
Natsch and
Emter (2008)

23. in vitro alternatives for safety testing
• European Centre for the Validation of Alternative
Methods (ECVAM)
• In 2013, ECVAM recommended the DPRA for use in
skin sensitization testing
• Direct Peptide Reactivity Assay (DPRA)
• Covalent binding of electrophilic chemicals to
nucleophilic centres in skin proteins is often a key
initiating event during the induction phase of skin
sensitisation
• DPRA mimic this step by quantifying the reactivity of
chemicals towards synthetic peptides containing either
cysteine or lysine as reactive core

24. DPRA evaluation

25. EpiDerm™ is 3-dimensional Human
Full Thickness Skin
• EpiDerm™ has been successfully utilized in
alternative in vitro toxicology tests for dermal
corrosivity, skin irritation, and phototoxicity
Stratum cornium
Epidermis
Dermis

27. A multi-endpoint, integrated testing approach that incorporates the
following end points:
• LDH to assess cytotoxicity
• GSH depletion assay to assess direct reactivity
• qPCR assays to detect up regulation of transcription of seven genes
involved in the Nrf2/Keap1/ARE or AhR/ARNT/XRE signalling
pathways
– NADPH-quinone oxidoreductase 1 (NQO1)
– Aldoketoreductase 1C2 (AKR1C2)
– Interleukin 8 (IL8)
– Cytochrome P450 1A1 (CYP1A1)
– Aldehyde dehydrogenase 3A1 (ALDH3A)
– Heme-oxygenase 1 (HMOX1)
– Glutamate cysteine ligase catalytic subunit C (GCLC)
SenCeeTox® Assay

28. NANOTECHNOLOGY
and
COSMETICS

29. Current Reported Uses of Nanotechnology
in Cosmetic Products
• Nanodispersed Systems
– Liposomes (nanosomes)
– Nanoemulsions
– Solid lipid nanoparticles
• Other Nanoparticles
– Polymer systems (nanocapsule, dendrimer)
– Metal Oxide Nanoparticles

30. Nanodispersed Systems
 Liposomes – lipid bilayer
 Nanoemulsion and lipid
nanoparticles – lipid monolayer
 Core can be hydrophilic or
hydrophobic depending on the
number of phospholipid layers
 Deliver water or lipid soluble
ingredients
Mufti, J.; Cernasov, D.; Macchio, R.; “New Technologies in Topical Delivery Systems” Happi.com, March 2002.

31. Cosmetic Nanoemulsions
• Nanoencapsulation of
ingredients
• May be found in skin and
hair care preparations
 Useful in typical
cosmetic formulations
 Light or oxygen sensitive
actives can be protected
 Vitamin A and Vitamin E
 Low biotoxicity of
phospholipid
US Patent 6335022; Simonnet; J._T., Sonneville; O., Legret; S.; L’Oreal, 1999.

32. Potential Lipid Nanoparticle
Advantages
• Improved stability of ingredients
• Controlled release of ingredients
• Improved skin hydration
• Incorporation of lipophilic ingredients
• No or low carrier biotoxicity

33. • Uptake-Mainly insoluble particles
• Should they become systemically
available, translocation/
transportation
• Accumulation in secondary target
organs may occur
Consideration on Use of nanoparticles
in Cosmetics
Scientific Committee on Consumer Products (SCCP-EU)-2007

34. Upper Hair Follicle

35. Image from: Toll
et al., J. Invest.
Dermatol. 123,
168-176, 2004.
Image shows penetration of microspheres into hair
follicle. Authors concluded that penetration was
between cuticula of hair shaft and inner root sheath
Laser scanning confocal microscopy (LSCM)

36. What Are Stem Cells
• Stem cells are
unspecialized cells
that have two defining
properties
– the ability to
differentiate into other
cells
– the ability to self-
regenerate
There are two types of stem cells, embryonic
and adult.

37. Role of Stem Cells in Skin regeneration
• human skin is the
largest
depository of
adult stem cells
in the body.
• in the basic layer
of the epidermis
• activated by
tissue injury or
disease.

38. Stem Cells in Cosmetics
U Autologous – self mesenchyme stem cell from abdomen fat---
growth factors
there are no actual stem cells in any cosmetic products

39. Plant Stem Cells
The source are the
meristematic tissues,
which are located in:
• root apical
meristem (RAM)
• shoot apical
meristem (SAM)
• and vascular system
((pro)cambium or
vascular meristem.

40. General Info
• Many stem cell cosmetics reside in a legal gray
area
• cosmetics do not require premarket approval
from the FDA- Is this applicable for Stem Cell
cosmetics?
• But stem cell cosmetics often satisfy the FDA's
definitions for both cosmetics and drugs
• What manipulation to cells in lab, what cell
product is used, what is the source of stem
cells???????

41. Regulatory

42. Summary
• Skin is Sensitive tissue
• Safety first
• Avoid using animal for testing
• Look for alternative techniques that can be
adopted by authorities
• Nanocosmetics have future in Market
• The notable market potential for stem cell –
based cosmetics obliges mature legislations