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Volume 14 Issue 1

 

96 well plate used in the KeratinoSens assay The KeratinoSens in vitro test method for skin sensitization testing, developed at Givaudan, has recently been evaluated by ECVAM. In summary, ECVAM found that the Keap1-Nrf2-ARE pathway is relevant for assessing skin sensitization. Having good predictivity and transferability, KeratinoSens is recommended to be used in combination with other assays (such as the Direct Peptide Reactivity Assay (DPRA)) in a Weight of Evidence (WOE) approach or as part of an Integrated Testing Strategy (ITS) for skin sensitization. Additional work is suggested by ECVAM to evaluate the assay's ability to predict potency categories relevant to humans. ECVAM states that "...data from the KeratinoSens test method should be considered in combination with complementary information in order to reduce and possibly avoid animal testing. As provided for in Annex XI (point 1.2) of the REACH Regulation (EC, 2006), data from non-standard testing methods, such as the KeratinoSens, may be used to adapt the standard information requirement in the context of WoE judgments."  

 

Late last year, ECVAM released their recommendation on the DPRA assay, developed at Procter & Gamble. They found that information from peptide reactivity assays, such as the DPRA, can be used for the assessment of the skin sensitization potential of chemicals, and that the DPRA is transferable to suitably equipped laboratories that are proficient in high performance liquid chromatography (HPLC) analysis. Although full review of the assay's predictive capacity was outside the scope of their review, its accuracy of distinguishing between sensitizers and non-sensitizers was 82%. The full text of the recommendations and summary data pdfs for both methods can be found by following the link above. 

SCIENCE
NociOcular Assay: A Novel In Vitro Assay to Assess Eye Stinging Potential        

   

The NociOcular Assay, developed by Dr. Anna Forsby at Stockholm University, is a novel neuronal model with high expression of functional TRPV1 channels.  The TRPV1 channel is a well characterized pain receptor that is expressed in sensory nociceptors, which can be activated by chemical stimuli.  Corneal and mucosal tissue in conjunctiva are rich in innervations which express TRPV1 channels. Therefore, TRPV1 channel activation is thought to be a general mediator of chemically induced pain on the surface of the eye.

 

In this cellular model, a TRPV1 expressing clone of the human SH-SY5Y neuroblastoma cell line was obtained by stable transfection. TRPV1 channel activation is measured by acute increases in the intracellular free Ca2+ using the fluorescent probe Fura-2AM.  Prior to Ca2+ measurements the TRPV1 expressing SH-SY5Y cells are cultured in 96-well plates.  The mean value (% increase of basal Ca2+   level) from triplicate wells is then monitored for each concentration from each independent experiment.  The TRPV1 antagonist capsazepine is added simultaneously with each concentration to confirm the specificity of TRPV1-mediated Ca2+  influx.

 

Although several in vitro eye irritation models exist, none of them has demonstrated the ability to predict the human sting potential of products which may come in contact with the eyes.  Therefore, an assay capable of detecting eye sting potential would be a beneficial pre-clinical screening tool. IIVS participated in a collaborative study with Johnson & Johnson Consumer and Personal Products Worldwide and Dr. Forsby's laboratory to test 19 baby cleanser formulations in the NociOcular Assay and compare the results to existing human clinical eye sting data. Our data, published in Toxicological Sciences, support that the TRPV1 channel is a principle mediator of eye stinging sensation induced by baby bath and shampoo formulations and that the NociOcular test may serve as a simple bioassay to ascertain this sensory response in the eye.

 

IIVS is pleased to introduce the NociOcular Assay as a new addition to our suite of in vitro assays. The current prediction model for the assay is based on studies with surfactant ingredients and formulations, but we seek to expand the applicability of the assay by performing additional investigation into other product types. If you would like more information about the NociOcular assay, please contact Dr. Kimberly Norman or Dr. Anna Forsby. 
EDUCATION
Practical Methods for In Vitro Toxicology Training Workshop and Custom Training Program for BTBU Scientists 
Click on the link below to register.
 
A key educational program at IIVS is our annual training course, Practical Methods for In  Vitro Toxicology. For over 17 years IIVS has instructed scientists from industry and government/regulatory and academic institutions on key laboratory techniques for conducting in vitro assays, and has assisted them in learning how to interpret the resulting data. During the multi-day program, participants are exposed to a variety of in vitro methods through lectures and hands-on activities with our highly trained biologists and Study Directors.
 
Held in January each year, the course features hands-on instruction on the Bovine Corneal Opacity and Permeability Assay (BCOP), 3T3 Phototoxicity assay, and use of 3D tissue constructs for dermal irritation testing. Participants are provided with demonstrations of a number of other assays during the 3 and a half day course including the KeratinoSens assay for skin sensitization testing, the Cytosensor Microphysiometer assay for eye irritation testing, and the Corrositex assay for determining DOT packing groups and hazard classification. Lectures on the assays include case studies to discuss data interpretation. Presentations on global acceptance of in vitro methods, the use of good laboratory practices, the US Tox 21 program, and new technologies for organ cultures round out the program. The success of this course has led to requests from individual companies and organizations for IIVS to create custom-designed workshops for their specific needs.

One such prog
ram was held in February when IIVS staff hosted two visitors from Bejing's Technology and Business University (BTBU) for an extended training on in vitro assays. BTBU is one of the few universities within China that has a cosmetics science academic program. One BTBU professor and a graduate student traveled to our facility to receive extended hands-on training in best practices for cell culture and non-animal toxicology assays such as the Bovine Corneal Opacity and Permeability assay (BCOP) and phototoxicity assay using 3T3 cells. During their time at our laboratory, they were also given training in the use of 3D tissue constructs for determining ocular and dermal irritation of chemicals and observed the performance of the KeratinoSens assay for determining skin sensitization potential. In addition to the hands-on performance of these assays in the laboratory in real time, they also received instruction on laboratory administration and spoke with our Study Directors about practical concerns such as equipment maintenance, ordering supplies, scheduling, and compliance with Good Laboratory Practices. IIVS gratefully acknowledges PETA for supporting this program through which a training laboratory at BTBU will be established for non-animal testing methods for cosmetics. IIVS staff will continue to provide remote support to the university as they launch their new program and open the laboratory.
  OUTREACH
International Cooperation - EPAA

 

In the fall of 2012, IIVS and the European Partnership for Alternative Approaches to Animal Testing (EPAA) signed a Memorandum of Understanding to coordinate efforts in progressing the international use of non-animal testing methods. The two organizations agreed to combine resources and collaborate to promote international awareness and education of these methods, and to provide science-based advocacy to key stakeholders. Since the signing of the MOU, IIVS has engaged in several activities in China which provided direct contact with scientists and key decision makers on the implementation of in vitro methods.

 

Dr. Quanshun Zhang, Manager of International Outreach at IIVS, attended the International Congress of Toxicology (ICT) in Seoul, Korea to interact with a delegation of scientists from China. These scientists came from major areas of government and academic institutions including the CFDA, CDC, Ministry of Environmental Protection, Chinese Academy of Science and Sun Yat-Sen University. Korea, with the formation of the Korean Center for the Validation of Alternative Methods (KoCVAM), serves as a model for the adoption of OECD Test Guideline methods and participation in international validation efforts. An "Alternatives Methods Section" of the Congress featured speakers from JaCVAM, ICCVAM and ECVAM who further reinforced advantages of international harmonization of alternative methods.

  

At the Chinese Society of Toxicology (CSOT) in Guangzhou, China, Dr. Zhang presented a poster on behalf of EPAA titled "International Sharing of Scientific Knowledge in Affecting Change in Regulatory Testing Approaches". Over 1,500 toxicologists attended the meeting from industry, government and academic/research institutions. As China considers changes to its regulatory testing requirements it is critical to discuss the advantages of in vitro testing approaches with key decision makers and thought leaders within China.

 

In addition to annual conferences such as the ICT and SOT, IIVS represented EPAA's commitment to international outreach at less formal venues designed to introduce industry scientists and students to in vitro methods. For example, Dr. Zhang was invited to speak at the Skin Biology Symposium organized by the Beijing Daily Chemical Association; an organization comprised of domestic cosmetic and ingredient manufacturers. The presentation focused on the application of 3D skin models in safety and efficacy assessment of cosmetics and ingredients. Dr. Zhang also provided a lecture to graduate students at the Beijing Technology and Business University (BTBU). See more about BTBU in the previous story. 

 

To broaden general awareness of alternatives in China, EPAA helped support the translation into Chinese and publication of The Three Rs and the Humanity Criterion by Prof. Michael Balls. This edition is an abridged version of The Principles of Humane Experimental Technique by William Russell and Rex Burch. This newly translated book will be distributed free of charge to many libraries and all major universities within China.

  

Recent Publications
New book published: Reducing, Refining and Replacing the Use of Animals in Toxicity

In the words of editors David Allen and Mike D. Waters, the text describes the ever-expanding "toolbox" of test methods available to toxicologists. By combing in silico, in vitro, and ex vivo methods toxicologists are moving closer to using mechanistically based alternatives without requiring animals.  

 

Chapters include the history of the 3Rs, regulatory testing, international harmonization, refinement, computational modeling, skin sensitization and more. The chapter titled "In Vitro Toxicology Models for Acute Eye and Skin Irritation Assessment" was authored by G. Costin and H. Raabe of IIVS. View portions of the book here.

Upcoming Events

March 25, 2014

Phoenix, AZ

 

March 24, 25, and 27, 2014

Phoenix, AZ

 

March 27, 2014

Phoenix, AZ

 

April 1-3, 2014

Hamburg, Germany

 

ToxCast Stakeholder Workshop 

April 02, 2014

Silver Spring, MD  

 

April 04, 2014

Cincinnati, OH

 

April 23, 2014

Languedoc, France

 

May 04, 2014

Quebec City, Canada

 

May 11, 2014

Basel, Switzerland

 

May13-14, 2014

Edison, NJ 

News
World Congress 9 Abstract Submission and Registration Opened


The 9th World Congress on Alternatives and Animal Use in the Life Sciences will be held in Prague, the Czech Republic, on August 24-28, 2014. The WC9 provides a forum to examine the importance of both the ethical issues of animal experimentation and the scientific approach to the life sciences in the 21st century. Participants are encouraged to contribute to the program by submitting abstracts, becoming a meeting sponsor, exhibiting, and registering as an attendee. Please follow this link to the WC9 website for more information.


Dr. Warren Casey Appointed Director of NICEATM


For the past year Dr. Warren Casey served as Acting Director of the NTP Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM) within the Division of the National Toxicology Program at the National Institute of Environmental Health Sciences. On January 7th Dr. John Bucher, Associate Director of the National Toxicology Program, announced that Dr. Casey has agreed to serve as the next permanent Director of NICEATM. IIVS wishes Dr. Casey well with his continued efforts at NICEATM, ICCVAM and ICATM. 
 

Visit Us at SOT
Booth#1027

March 24-27

Phoenix, AZ 

 

 
The Society of Toxicology provides an excellent forum to meet with IIVS staff to discuss in vitro testing strategies. Please visit us in the ToxExpo at booth 1027 or email us now to set up an appointment time. IIVS Study Directors will also be presenting several posters during the meeting. 

 




IIVS at In-Cosmetics
Stand 4H17 

April 1-3

Hamburg, Germany 

 

Join IIVS staff at In-Cosmetics, the leading global business platform for personal care ingredients. IIVS is pleased to sponsor the workshop "The Animal Testing Ban: What's Next". Please visit us in the Testing Zone at stand 4H17 to discuss the latest in non-animal testing strategies. 




NYSCC Supplier's Day 

May 13-14
New Jersey Convention Center
Edison, NJ

Please visit IIVS Study Directors at the upcoming Supplier's Day to discuss your programs and how our new technologies can be utilized to meet your testing needs.

     

Would you like to learn more about in vitro skin sensitization assays? Send us an email at
 +1 301 947 6523 
 

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ATLA Q&A with IIVS, 2012 LUSH Cosmetics Prize Winners for Training  
 
IIVS is pleased to be featured in a Question and Answer article in ATLA Journal 41. The focus of the article is our 2012 LUSH prize award for training activities. The article discusses the background of the Institute, why we were nominated for the prize, as well as the impact the prize had on our training work last year. To read the article, please follow this link to the FRAME website.



 


Bill to End Animal Testing of Cosmetics Proposed in the US  Congress




 The "Humane Cosmetics Act" (H.R. 4148) was introduced in the US Congress in March. According to the Humane Society of the United States, this act would 1) Make the conduct or commission of animal testing of cosmetics unlawful in the U.S., and 2) Prohibit sale or interstate transport of cosmetics if the final product or any component was developed or manufactured using animal testing. For more information on the bill, please follow this link. 
  


New Tutorials for the Use of the OECD QSAR Toolbox Version 3.2 are Available 

 

 

 Four new tutorials are available that provide illustrated examples for the use of the functionalities of the Query tool , the AOP workflow for Skin Sensitization, the information from the ECHA CHEM database and the capability to analyze tautomeric structures. The examples cover both human health (skin sensitization) and environmental (acute aquatic toxicity to fish) endpoints.   
  



So Paulo became the first Brazilian state to ban the use of animal testing to determine the safety of cosmetic products and ingredients. The bill, which was signed into law in January, includes specified fines for violations of this law. India has also resolved to end animal testing for household products. This recent ban comes after last year's move to end tests on animals for cosmetics and their ingredients by removing animal tests from the relevant Bureau of Indian Standards. 

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American Society for Cellular and

Computational Toxicology

 

 

   

ASCCT at the  

 

Society of Toxicology Annual Meeting
Stop by the Global Gallery of Toxicology on Monday, March 24,  between 11:45-12:15, in the ToxExpo Exhibit Hall. ASCCT will be presenting a poster on our mission and activities. It's a great opportunity to meet other members and show support for the society.

 

  

Don't forget to renew your membership for 2014 to continue to receive the informative ASCCT newsletter and free access to webinars!
 

www.ascctox.org