Addressing Nano in Your FoodA model for multistakeholder science

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Overview

I) Challenges to developing public health science for food safety

II) A project model that can help address these challenges

III) Case example: NanoRelease Food Additive project

IV) State of the Science on nano in food (Dr. Andrew Bartholomaeus)

V) Conclusions & specific next steps, open discussion (feedback on this approach)

Challenges in public health science regarding food safety

1) Diminishing public and private funds

2) More disruptive “politicization” of the science

– Polarization – Us vs Them– Immediate broad access to

information means that more of the uncertain hazard information makes it into general awareness

3) More technical complexity

• Explosion of new data types – For assessing biological effect– For monitoring

• Technologies in food supply– Nanotech, biotech– Personalized nutrition, functional foods

• Information technology– Sheer data volume and need for new decision approaches– Data integration (in silico, computational tox)

4) Failure of the top down or single source approaches to evaluation

Bisphenol-A example of how not to develop public policy from science• Health authorities evaluated risks and made announcements • Some members of the public agreed with health authorities,

some did not• Press attention to the debate affected perceptions • Decisions were made in public and in private arenas• In the end it seems all sides are asking whether the science

was used appropriately and whether public health is improved

How can we address these challenges and move forward effectively?

We need to publish or produce data in ways that are good science and do not have

– Single stakeholder bias– Labels as being from “one side” of the debate

The proposed approach is to• Assemble expert stakeholders from opposing views• Build trust between them & establish

continuous dialogue • Move forward toward a goal

Formulate the issue in a way that addresses public health from multiple perspectives

Gather senior experts to form the initial charge for the project (multi-stakeholder, multinational)

Make it clear that they control the project outcome, all proceedings are public, and that the outcomes (publications,

methods) will be public or the intellectual property of the experts of the project.

The core group invites more participants through their networks, becomes the Steering Committee with co-chairs rotating across

stakeholder groups and develops the plan of work.ILSI RF becomes the Secretariat supporting the Steering Committee.

Structure & Flow of project developmentStep 1: Intellectually invest experts from opposing perspectives in the outcome

The core group is made up of risk managers, not technical experts.

Steering Committee Invites Experts to Task Groups

Step 2: Build a transparent project run by the experts

Secretariat coordinates Task Groups to address their charges

Secretariat raises funds to support the project, convenes meetings, takes notes of meetings, records project activities and outcomes on a project web site, supports workshops, aids in development of documents as needed.

Step 3: Make the outputs independent of the funders

Steering Committee identifies expert subtopics to be evaluated by Task Groups

(to inform selection of a solution approach)

Steering Committee (risk managers) use the information of the Task Groups to inform “state of

the science” or other evaluations regarding the issue.

Task groups independently publish white papers or develop outputs (e.g., methods)

Task Group experts are invested in the outcome through publications they produce.

State of the Science or other evaluations are also independent publications by authors of the group.

Incentives for participants• Project participants own and run the project• Leveraged funding

– No single entity funds more than 10% of the project

• Publications• Extended science policy discussions between stakeholders

– Builds trust– Time to talk through the issues (in a trusted forum)

• Extended access to key experts in the field• Access to developing science • Publications that fill in gaps in understanding

Government54%

Industry28%

Academics15%

CSO3%

Effort by Stakeholder Entity

Industry 201141%

Government 20116%

Government 201236%

Industry 201216%

Funding 2011-2012

Steering Commit -tee 31%

Task Group White Papers37%

Task Group Partic-ipant7%

Workshop partic-ipation

32%

Functional Breakdown of Time Spent Public-Private PartnershipsIt’s more than the money

~20 experts on Steering Committee~50 on 4Task Groups

Lower bound estimate of time based on meeting minutes

Using a rough estimate of $120/hr, the total value is about $1M

NanoRelease Consumer Products

Example of agreement across stakeholders

We need to know what is in consumer hands so that we can design the best methods to assure safety

There is no good information about what products use nanomaterials/nanotechnology

The task group experts identified the most used materials based on their own experiences and from purchased market surveys

Because we have the experts who are making the materials in industry and who are reviewing the product authorizations at government, we have the best unpublished current information

Example of agreement across stakeholders

The steering committee is using the project to develop methods to measure what is coming off of real products

We need to know what is really coming off of consumer products and test it in toxicity studies

Existing hazard data is generally not related to consumer exposure

Example of agreement across stakeholders

We need to a) focus attention on where

there is higher release and b) have trusted methods for

proving the lack of release elsewhere

Most of the uses of nanomaterials are safe (because very little is released)

Project experts submitted a “life cycle release scenarios” evaluation that identifies where releases are likely and where proving lack of release is needed

The Steering Committee is using this knowledge to develop the best methods to measure what is coming off of real products

More detail on a case example…NanoRelease Food Additive project

FOOD SAFETY TOPIC: Nanotechnology in food

CHALLENGES (as stated previously): 1) Diminishing public and private funds

2) More disruptive “politicization” of the science

3) More technical complexity

4) Failure of the top down or single source approaches to evaluation

State of the science on nano in foodDr. Andrew Bartholomaeus

THE ORAL PHARMACOKINETICS OF PARTICLES

Dr Andrew BartholomaeusProfessor (Adj)

Therapeutic Research Unit, School of Medicine, University of Queensland

School of Pharmacy, University of CanberraBartCrofts Scientific Services

If you increase the magnification

another million times you can see the

safety regulations.

First we need to understand What are we talking about

Current definitions do not separate new technology from old

Provide no regulatory or research target Obscure pre-existing knowledge Largely unhelpful

What makes nanoparticles different?

They are small They might go where larger particles cannot Protein complexes can be of equivalent size Biological interactions bridge the chemical/physical

boundary Surface area: volume ratio is large

Surface reactivity is high Solubility increases

Curvature is high Influences intermolecular interactions

Potential for engineered novelty Most of the above really only apply at < 20 or 30

nm and significant around 5 nm

HOW NOVEL ARE THEY

What we can learn from past science

Forró L and Schönenberger C. Carbon nanotubes, materials for the future. Europhysics News 2001;32(3).

Old wine in a new bottle ? Those who cannot remember the past are

condemned to repeat it1. (1905) (Colloids)…..this fascinating twilight zone

between physics and chemistry2. (1919) Much of what is presented as new

technology is in fact an extension of long standing knowledge and practices, albeit considerably more sophisticated.

1. George Santayana, Life of Reason, Reason in Common Sense, Scribner's, 1905, page 2842. Jerome Alexander, Colloid Chemistry, D.Van Nostrand Co, NY, 1919

“…extending the sphere of interest in this fascinating world between physics and chemistry.”

Micelles and Emulsions1 The Pharmacists Engler and Dieckhoff discovered they

could produce clear solutions of cresol in concentrated aqueous solutions of soaps in in 1898.

.. the small molecules in dilute solutions tend to associate into aggregates (micelles) of equivalent diameters in the 30 to 100 Å (3-10 nm) range….they are called association colloids. Other names are surfactants or surface active agents.

1Remington’s Pharmaceutical Sciences. 15th Ed, Mack Publishing Company, Easton, Pennsylvania. 1975

MICELLES ARE DYNAMIC, SELF ASSEMBLING NANO-STRUCTURES

Material Food Product Size (nm)All polysaccharides Edible plant and muscle tissues,

milk, eggs, processed foods~50–1500

Glycogen Edible muscle tissue and liver 8–43Starch granules’ internal concentric rings

Edible plant tissues 100–400b

Starch granules’ amylopectin clusters Edible plant tissues 5–10

Unsaturated triglyceride Vegetable oils ~3Cholesterol Animal lipids ~1.5Myosin Edible muscle tissue 1.5–2 diameter, 100 in

lengthCollagen Edible muscle tissue 1.4- to 1.5-wide unitsWhey Milk 4–6Enzymes Naturally existing or added 1–10A, D, E, K, C, thiamin, riboflavin, niacin, B6, B12, biotin

Naturally existing or added <1–2

Lycopene Tomatoes ~3Beta-carotene Carrots, oranges, peaches, peppers ~3Capsaicin, gingerol, tumerone Capsicum peppers, ginger, turmeric ~1–2Casein micelle Raw milk 30–300

Many food components fit the nanoscale definition

Food related applications of nanotechnology

Parrots regularly eat seeds and unripe fruits whose content of alkaloids and other toxins renders them bitter and even lethal to humans and other animals. Because many of these chemicals are positively charged in the acidic conditions found in the stomach, they bind to clay minerals bearing negatively charged cation-exchange sites...“(Jared Diamond)

Food = Matter Food naturally & traditionally contains particles in the nanometre scale

Uses of nanoparticles in food Traditional

Silicon dioxide (E551) Homogenised milk (200 to 2000 nm)

New or proposed uses Nanoencapuslated nutraceuticals: vitamin E,

CoQ etc Nanoencapsulated preservatives Nanoliposomes in cheese manufacture Nanoclusters to enhance flavour of a chocolate

slimming drink Phytosterols in canola oil

Electron micrographs of human breast milk showing casein micelles following centrifugation. Bar = 1 µm in B and 0.2µm in C. [1]

Fat globules in ice cream. (A) entire globule with coating of casein subunits, (B) broken globule with collapsed coating of casein subunits, (C) crater left behind by a fat globule, showing casein subunits aligned around periphery. [2]

Extruded ice cream [2]

1From: Keenan & Patton. The Structure of milk: Implications for sampling and storage. In, Handbook of milk composition. R.G. Jensen ed. Academic Press. 1995

2. K.G. Berger. Ice Cream. In Food Emulsions. Stig Freiberg ed. Marcel Decker, NY. 1976, 141-210

Nanostructures in food

Aerosil – fumed silica dioxide, used in various food, cosmetics, paints and pharmaceutical applications for over half a century Technical Bulletin, Fine Particles, Basic Characteristics of AEROSIL® Fumed Silica Number 11

https://www.aerosil.com/www2/uploads_all/text/SR_11_AE_us_Basic_Characteristics_of_AEROSIL_2006-04.pdf

Key Science IssuesPharmacokinetics

Pharmacokinetics Key potential novelty of

nanomaterials Potential for transitional behaviour

between particles and chemicals Size alone is not a sufficient metric

to predict pharmacokinetics

Absorption of Particles – Is it novel ?

Direct “persorption” of µm size particulates (15-75 µm optimum) across the GI tract wall was first observed in 18441, but some scepticism remains

Particles of starch, charcoal, sulphur, rabbit hair, silica, etc were variously studied in rabbits, dogs or frogs and found to be taken up into blood, bile and urine.

Transport from site of persorbed particles is via chyle or portal blood.

Absorption pathways for nanoparticles may differ to that of microparticles but their absorption is not novel per se.

1. Volkheimer, G., (2001) The phenomenon of persorption: persorption, dissemination, and elimination of microparticles. In: Old Herborn University Seminar Monograph. 14. Intestinal Translocation. ISBN 3-923022-25-5

2. Volkheimer, G (1974) Passage of particles through the wall of the gastrointestinal tract. Environmental Health Perspectives. Vol 9, 215-225

Absorption of nano and micro particles is Normal

Particulate Persorption as a function of size

Florence, T. (1997) The oral absorption of micro and nanomaterials: Neither exceptional nor unusual. Pharmaceutical Research, Vol 14, No 3, 259-266

Size dependent routes of absorption

1. Persorption Very large particles (5-75 microns) – some contention

here Quantitatively small proportion but numerically

significant2. Uptake by M cells of the Peyer’s patches

Direct phagocytic immune sampling of gut contents Favours particles around 1µm Uptake of particles of smaller or larger dimensions

appears less specific3. Transcytosis through enterocytes

Generally only significant for nanoparticles with specific ligands promoting receptor mediated endocytosis

Absorption of 500 nm polystyrene beads in rats increased 50 fold by coating with tomato lectin

4. Paracellular transport Paracellular pores < 1% of luminal surface Pore size approx 1 nm in size

Uptake 10 into Lymphatic Sysem not Hepatic Portal Vein

Optimum size for lymphatic transport 10-100 nm (Swartz in Advanced Drug Delivery Reviews 50 (2001) 3–20)

Molecules that are smaller than 10 nm are preferentially reabsorbed into the blood capillaries

Effect of size and shape on endocytosis

Chithrani, B.D. and Chan, W.C. (2007) Nano.Lett 7(6):1542-1550.

Intracellular uptake by HeLa cells, as measured using ICP-AES (inductively coupled plasma atomic emission spectroscopy)

Tissue distribution

Zhang, G., Yang, Z., Lu, W., Zhang, R., Huang, Q., Tian, M., Li, L., Liang, D. and Li, C. (2009) Biomaterials 30(10):1928-1936.

Tissue distribution of 20 and 80 nm pegylated gold nanoparticles in nude mice with human squamous carcinoma A431 SC (3/group) 48 h post IV administration

Ogawara, K., Furumoto, K., Takakura, Y., Hashida, M., Higaki, K. and Kimura, T. (2001) J Control Release 77(3):191-198.

Driving issue: Disagreement on what is nano in foods and whether we are addressing risks

Focus: What is the state of the science on methods to measure the oral uptake of nanomaterials?

Goal: Establish a widely-accepted set of methods for measuring oral uptake of nanomaterials

NanoRelease Food Additive project

Early 2012: Steering Committee

– Clarified scope, developed task group charges, recruited experts.

– Created extensive list/database of relevant studies and projects (80+ references gathered)

– Identified nanomaterial characteristics of interest through extensive deliberation and expert input

Task Group 1: MATERIAL CHARACTERISTICS What do we need to know about the nanomaterials and the food matrices to predict absorption as particles into the body?

Task Group 2: ALIMENTARY CANAL ENVIRONMENT What do we need to know about alimentary tract conditions to understand whether and where a nanomaterial will be absorbed into the body?

Task Group 3: ALIMENTARY CANAL MODELSWhat kinds of models are useful in creating the conditions to measure and understand nanomaterial uptake by the body?

Task Group 4: MEASUREMENT METHODSWhat methods can be used to measure characteristics of materials to understand and predict nanomaterial uptake by the body?

Task Group 5: RISK MANAGEMENT CONTEXT Where in the decision process do we most need agreement to such measurement methods?

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NanoRelease Food Additive Sponsors• The Pew Charitable Trusts

• US Food and Drug Administration

• ILSI North America, Food and Chemical Safety Committee

• Illinois Institute of Technology’s Institute for Food Safety and Health

• Health Canada

• The Coca Cola Company

• Substantial in-kind support is provided by the Nanotechnology Industries Association

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Participants & Supporting OrganizationsILSI branches are participating actively (invited, consulted, included)

USACanadaBelgiumGermanyIrelandItaly

NetherlandsPortugalScotlandSpain

Switzerland

UKJapan

South Korea

Australia

Arizona State University Leitat Technological Center University of FloridaAs You Sow Louisiana State University University of GuelphCargill Michigan State University University of GuelphCentre For BioNano Interactions MRC Human Nutrition Research University of IllinoisColorCon, Inc. Nanotechnology Industries Assoc. University of MarylandConsumers Union Ohio State University University of Massachusetts

Cornell Food SciencePeople for the Ethical Treatment of Animals University of Michigan

DSM Nutritional Products PepsiCo University of MissouriEuropean Commission JRC PerkinElmer Company University of South AlabamaEvonik Industries Purdue University University of TennesseeFera UK RIKILT - Institute of Food Safety University of TorontoFriends of Earth Rutgers UniversityGE Global Research Saarland University US Department of Agriculture

Health Canada Southwest Research Institute US Food and Drug Administration

Heriot-Watt University Taiyo Kagaku US National Cancer Institute

Hoseo University The Pew Charitable TrustsUS National Institute of Standards and Technology

IIT Institute for Food Safety and Health TNO NetherlandsILSI Europe University of California Davis

ILSI North AmericaUniversity of Canberra & University of Queensland

Istituto di Chimica e Tecnologia dei Polimeri University of East Anglia

Materials in commerce

Regulatory definitionTargeted Nanomaterials

Unintended Nanomaterials

Regulators dilemma: a) we don’t want to miss anything

b) we don’t want to add new regulation to innocuous materials

Are we ready to re-regulate all the materials that will be roped in?

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Are there simple ways of reducing the infinite class of “between 1 and 100 nm” to the risks we are concerned about for oral exposure (through foods)?

Proposal: A good start to getting our arms around risks is to see if we can identify the nanomaterials that are likely to be absorbed as particles into the body.

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Copied from ILSI Europe guideline

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Hypothetical approach for food nanoparticle evaluation (to prioritize data needs or aid product development)

Is it soluble in gastric conditions in adult?

In infant? Disease states?

Do particles pass to systemic circulation in adult?

Age/disease variation?

Are particles found in tract lining cells in adult?

Age/disease variation?

If insoluble, does it aggregate/bind irreversibly to particles greater then 10

micron?

Decreasing relative

proportion of materials in commerce?

Increasing need to apply

nanoparticle specific toxicity

tests

Design products preferably in this range

Widely agreed to, robust methods allow sustainable product development and transparent evaluations

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As a product developer– Can I make it dissolve to non-toxic materials – If no, then can I make it agglomerate or bind

irreversibly to particles that pass without absorption?

– If no, then can I use a material that results in cell uptake below detection?

As a concerned stakeholder– Using a standard test, does the material dissolve?– Is uptake undetectable using standard tests?

Outcomes• Trusted dialogue of what is needed to inform safety

decisions

• Trusted, robust methods that all can use to develop comparable data

• Framework for applying methods that– Clarifies risk management and data development

decisions

– Enables safe product development

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Example of agreement across stakeholders

Some new processes intentionally create small particles for effect (nano) and we do not know if those particles are better absorbed by the body.

Lots of natural or traditional food components are nano (so much of the “nano” is what we have been doing for a long time).

Discussion and decision processes across groups has built trust and helped us systematically focus on what is important.

(diagram is from working draft document within NanoRelease Food Additive project)

Example of agreement across stakeholders

We don’t have good information about what is in foods.

Because we have the experts who are making the materials in industry and who are reviewing the product authorizations at government, we can access the best unpublished current information

Characteristics of InterestSC created a decision matrix to identify the nanomaterial characteristics of interest to the project.

Example of agreement across stakeholdersWe need to

a) focus attention on what makes it to the mouth as nano and

b) have trusted methods for identifying what stays as nano as absorbed

Nanotech may in fact be used at many stages in food processing and packaging, however the “nano” nature may not make it to the mouth or past the initial transformations in the gut (dissolution, aggregation, binding to food components).

Project experts are developing a decision tree approach to when measurement of nanoparticles will be useful to understanding whether they are absorbed by the body.

The Steering Committee is using this knowledge to select the best methods to develop to be able to measure…

What is the intended use and design intention? How is it engineered to be nano?

At what point of the food manufacturing process is it used?

What is the concentration and physical state of the nanomaterial at

ingestion?

Does the concentration of the nanomaterial in the food affect the

dissolution/agglomeration rates?

How does the nanomaterial change at each anatomical region of the alimentary canal (sequentially)?

Does its use change if from being a nanomaterial?

It is critical to ask when it is nano

Step 2: Build a transparent project run by the experts

Bright lights on all processes. Steering committee is the project manager.

Step 3: Make the outputs independent of the funders

Add a second step of distance from the funding.

Step 1: Intellectually invest experts from opposing perspectives in the outcome

Formulate issues in solvable, practical terms.Get all perspectives to the table early.

Approach to public private partnerships for divisive public health issues

Open discussion about the project structure (feedback on this approach)

1) Is this project approach useful?

2) How would you improve it?

3) What other topics could be applicable for addressing using this approach?