industry applications for 3d bio- printing - consumer products
TRANSCRIPT
INDUSTRY APPLICATIONS FOR 3D BIO-PRINTING - CONSUMER PRODUCTS
ANDREW SCOTT,SAFETY & ENVIRONMENTAL ASSURANCE CENTRE
www.TT21C.org
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WHAT I’LL COVER TODAY
• About Unilever
• New technology and lead identification
• Assuring consumer safety without new animal tests
• A perspective on 3D-Bioprinting and where the bioprinting community can help
• Case study – human relevant skin models
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“MAKE CLEANLINESS COMMONPLACE” 3
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EUROPE
● €13.2 BILLION TURNOVER
● 0.2% UNDERLYING VOLUME GROWTH
● 27% OF GROUP TURNOVER
ASIA, AFRICA, CENTRAL & EASTERN EUROPE
● €19.7 BILLION TURNOVER
● 2.0% UNDERLYING VOLUME GROWTH
● 41% OF GROUP TURNOVER
THE AMERICAS
● €15.5 BILLION TURNOVER
● 0.7% UNDERLYING VOLUME GROWTH
● 32% OF GROUP TURNOVER
2014 TURNOVER = €48.4 BN
UNILEVER ISA GLOBAL COMPANY
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WE MAKE MANY OF THE WORLD’S FAVOURITE BRANDS
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INGREDIENT DISCOVERY FOR PERSONAL CARE
Mechanistic Insight
Target ID & Validation
ScreeningIngredient
testing in vitro
Validation in human studies
Representative in vitro models needed
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MICROCIRCULATION AND THE SKIN
• Skin is highly vascularised:
• Superficial plexus - below the skin
surface
• Sub papillary plexus - at the dermal-
subcutaneous junction.
• A complex molecular dialogue exists
between vascular derived cells and
mesenchymal skin cells impacting on skin
function.
• 3D bio-printing will help the investigation of
complex signalling pathways and cross talk
mechanisms apparent in vivo.
Li et al 2006
65 years old,
hand
Hembold et al 2006
old
young
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THE HAIR FOLLICLE – A COMPLEX MINI-ORGAN
(Schneider et al., 2009 Current Biology)
The hair follicle undergoes regular
cycles regeneration throughout life.
Hair follicle is a complex mini-organ
of the skin.
The Philpott hair follicle organ
culture model relies on availability
of waste donor-consented scalp
tissue following surgery.
Currently the above or variations of
the Philpott model is the only whole
hair follicle in vitro model available
despite attempts to grow hair
follicles in co-culture systems.
3D bio-printing combined with iPSC
technology may provide a solution
for overcoming limited supply of
tissue and recapitulation of hair
follicles that mimic those in vivo
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DEVELOPMENT OF SCALP SKIN MODELS
Scalp Skin
Development of scalp skin modelsReconstructed Models
Epiderm – skin equivalent Scalp epidermal model
Measure barrier
integrity (TEER)+ ‘SEBUM’
+ MICROBES
(lab grown or ex-vivo)
Microbes removed
‘microbially challenged’epidermal model
T-cell priming
Measure protein and mRNA biomarkers of
immune response
Measure epidermal inflammatory
response (transcript)
Measure epidermal inflammatory
response (protein)Epiderm embeddedwith Dendritic cells
Measure protein and mRNA biomarkers of
immune response
+ immune components
epidermal
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Can we use x% of ingredient yin product z?
CAN WE USE A NEW INGREDIENT OR SAFELY?
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PERSONAL CARE CONSUMER PRODUCTS
• Chemical ingredients not generally intended to be pharmacologically active (compare Pharmaceutical Co.)
• Low bioavailability and often topical exposure
• Open regulatory environment
Making an exposure-led safety decision based on confidence that the safe level is within or below the adaptive homeostasis response, captured by appropriate in vitro systems and complemented with network computational models
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NEW APPROACHES TO RISK ASSESSMENT WITHOUT ANIMALS
• Focus on non-animal approaches for consumer safety risk assessment
• Data required for safety decision should be driver
• Dose response information is essential
• Understanding the underpinning human biology
• We are not looking for a way to do the animal test without the animal
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ASSURING CONSUMER SAFETY
Start with exposure rather than toxicity
‘ ……. Human safety depends on exposure and toxicity. Indeed, the 2012 European Commission report on addressing the NewChallenges for Risk Assessment states, “ A paradigm shift is likely from a hazard-driven process to one that is exposure-driven” ….’
Pastoor et al (2014), Crit Rev Toxicol, 44(S3): 1–5
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EXPOSURE – DRIVEN SAFETY ASSESSMENT
• Improving exposure estimates• Building on the current knowledge of consumer habits & practices• Focus on measurement and quantitative exposure science
Dermal kinetics
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WILL THESE EXPOSURES CAUSE ADVERSE OUTCOMES IN CONSUMERS?
BiologicInputs
NormalBiologicFunction
Adaptive StressResponses
Early CellularChanges
Exposure
Tissue Dose
Biologic Interaction
Perturbation
Low DoseHigher Dose
Morbidityand
Mortality
Cell Injury
Higher yet
(From Andersen & Krewski, 2009, Tox Sci, 107, 324)
Perturbations of Toxicity Pathways
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AN EXAMPLE PATHWAY
• Skin allergy
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Adapted from ‘Adverse Outcome Pathway (AOP) for Skin Sensitisation’, OECD
OPPORTUNITIES FOR BIO-PRINTING MAPPED TO SKIN SENSITISATION AOP
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OPPORTUNITIES FOR BIO-PRINTING MAPPED TO SKIN SENSITISATION AOP
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Replace ex vivo human skin for measurement of skin bioavailability (binding, metabolism,
penetration)
OPPORTUNITIES FOR BIO-PRINTING MAPPED TO SKIN SENSITISATION AOP
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Replace ex vivo human skin for measurement of skin bioavailability (binding, metabolism,
penetration)
Characterise impact of chemical/drug/product exposure on skin micro-environment
(e.g. induction of inflammatory response, interplay between skin microbiome and local immune cells)
OPPORTUNITIES FOR BIO-PRINTING MAPPED TO SKIN SENSITISATION AOP
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Replace ex vivo human skin for measurement of skin bioavailability (binding, metabolism,
penetration)
Characterise impact of chemical/drug/product exposure on skin micro-environment
(e.g. induction of inflammatory response, interplay between skin microbiome and local immune cells)
Develop skin models recreating different skin
disease pathologies to inform ‘sensitive’ sub-population
risk assessments
OPPORTUNITIES FOR BIO-PRINTING MAPPED TO SKIN SENSITISATION AOP
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TKTD MODEL OF HUMAN SKIN SENSITISATION: SCHEMATIC DIAGRAM
PANEL A = TOXICOKINETIC MODEL: DNCB kineticsin skin due to (1) diffusion and partitioning intothe stratum corneum and skin; (2) sensitiserclearance by dermal capillaries; (3) covalentmodification of protein nucleophiles by hapten.
PANEL B = TOXICODYNAMIC MODEL: biological response due to (4)proteasome processing of protein nucleophiles to form small peptides andtransport to the endoplasmic reticulum (ER); (5) binding of peptides andhapten-peptide complexes to MHCI and transport to plasma membrane; (6)binding of pMHC and hapten-pMHC to CD8+ T cell receptors and (7)activation and expansion of naïve specific CD8+ T cells.
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SUMMARY – OPPORTUNITIES FOR 3D BIO-PRINTED (SKIN) MODELS
• Improved speed and accuracy for identification of lead actives
• More predictive tissue models for investigate research and target identification
• More biologically relevant models for assessing effects on pathways
• Potential replacement for limited human tissue supply
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Unilever’s Safety & Environmental Assurance Centre (SEAC): helping to shape innovations that are safe for our consumers and workers, and better for the environment. SEAC was created 25 years ago by bringing together all relevant scientific expertise across Unilever in a single group.
1990 – 2015
AOP-BASED RISK ASSESSMENTSEXAMPLE: SKIN ALLERGY
Induction of skin allergy is a multi-stage process driven by toxicity pathways
- mechanistic understanding is captured in Adverse Outcome Pathway (AOP)
- non-animal test methods have been developed; each aims to predict impact of a chemical on one key event
- how can we make risk assessment decisions by integrating this scientific evidence?
Modified from ‘Adverse Outcome Pathway (AOP) for Skin Sensitisation’, OECD report
1. Skin Penetration
2. Electrophilicsubstance:
directly or via auto-oxidation or metabolism
3-4. Haptenation: covalent
modification of epidermal proteins
5-6. Activation of epidermal
keratinocytes & Dendritic cells
7. Presentation of haptenated protein by
Dendritic cell resulting in activation &
proliferation of specific T cells
8-11. Allergic Contact Dermatitis: Epidermal inflammation following
re-exposure to substance due to T cell-mediated cell
death
Key Event 1 Key Event 2 + 3 Key Event 4 Adverse Outcome