ATLA 40.1, March 2012

//ATLA 40.1, March 2012

Volume 40 Issue 1

Editorial: China is Taking Steps Toward Alternatives to Animal Testing

Rodger Curren and Brian Jones

The rate at which China is addressing the problems associated with animal testing may be reasonable, when one considers the fact that the Three Rs concept of refinement, reduction, and replacement of animal tests to detect chemical hazards, which is relatively new to China, has already existed for over 50 years in most English-speaking countries. But for only about half that time has it been considered by a majority of scientists as a respectable goal to pursue.
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News & Views

ATLA Staff Writer

UK Biobank Opens
InterNICHE Launches New Website
Resignation of Director of Harvard Primate Center
A Model of Human Implantation
Empathically-motivated Behaviour in Rats
An Alternative to Rat Caries Testing
ICCVAM Recommends In Vitro Test Method for Endocrine-disruptors
Microfluidic Technology Used to Model Haematologic Diseases
Grant for Development of Replacement Methods
International Transport of Laboratory Animals
Debate on Transposition of Directive 2010/63EU
Ban on the Use of Great Apes
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CAAT News & Views

ATLA Staff Writer

Food Information Day
CAAT-Europe and ECOPA Co-operation
Refinement Working Group
The Human Toxicology Project Consortium
National Academy of Sciences Report on Animal Models for Assessing Countermeasures to Bioterrorism Agents
CAAT Activities in India
Evidence-based Toxicology Collaboration Programme
Recent publications by CAAT/CAAT-Europe Faculty
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IIVS News & Views

ATLA Staff Writer

New Director of Education and Outreach Programmes
Cosmetic Industry: Looking into the Future
International Outreach Programme: China
IIVS Practical Workshop for Alternative Methods in Brazil
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Testing the Biocompatibility of a Glutathione-containing Intra-ocular Irrigation Solution by Using an Isolated Perfused Bovine Retina Organ Culture Model — an Alternative to Animal Testing

Kai Januschowski, Ahmad Zhour, Albert Lee, Ramin Maddani, Sebastian Mueller, Martin S. Spitzer, Sven Schnichels, Maximilian Schultheiss, Deshka Doycheva, Karl-Ulrich Bartz-Schmidt and Peter Szurman

The effects of a glutathione-containing intra-ocular irrigation solution, BSS Plus®, on retinal function and on the survival of ganglion cells in whole-mount retinal explants were studied. Evidence is provided that the perfused ex vivo bovine retina can serve as an alternative to in vivo animal testing. Isolated bovine retinas were prepared and perfused with an oxygen-saturated standard irrigation solution, and an electroretinogram was recorded to assess retinal function. After stable b-waves were detected, the isolated retinas were perfused with BSS Plus for 45 minutes. To investigate the effects of BSS Plus on photoreceptor function, 1mM aspartate was added to the irrigation solution in order to obtain a-waves, and the ERG trace was monitored for 75 minutes. For histological analysis, isolated whole retinal mounts were stored for 24 hours at 4°C, in the dark. The percentages of cell death in the retinal ganglion cell layer and in the outer and inner nuclear layers were estimated by using an ethidium homodimer-1 stain and the TUNEL assay. General swelling of the retina was examined with high-resolution optical coherence tomography. During perfusion with BSS Plus, no significant changes in a-wave and b-wave amplitudes were recorded. Retinas stored for 24 hours in BSS Plus showed a statistically significant smaller percentage (52.6%, standard deviation [SD] = 16.1%) of cell death in the retinal ganglion cell layer compared to the control group (69.6%, SD = 3.9, p = 0.0031). BSS Plus did not seem to affect short-term retinal function, and had a beneficial effect on the survival of retinal ganglion cells. This method for analysing the isolated perfused retina represents a valuable alternative for testing substances for their retinal biocompatibility and toxicity.
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Validation Study of the In Vitro Skin Irritation Test with the LabCyte EPI-MODEL24

Hajime Kojima, Yoko Ando, Kenji Idehara, Masakazu Katoh, Tadashi Kosaka, Etsuyoshi Miyaoka, Shinsuke Shinoda, Tamie Suzuki, Yoshihiro Yamaguchi, Isao Yoshimura, Atsuko Yuasa, Yukihiko Watanabe11 and Takashi Omori

A validation study on an in vitro skin irritation assay was performed with the reconstructed human epidermis (RhE) LabCyte EPI-MODEL24, developed by Japan Tissue Engineering Co. Ltd (Gamagori, Japan). The protocol that was followed in the current study was an optimised version of the EpiSkin protocol (LabCyte assay). According to the United Nations Globally Harmonised System (UN GHS) of classification for assessing the skin irritation potential of a chemical, 12 irritants and 13 non-irritants were validated by a minimum of six laboratories from the Japanese Society for Alternatives to Animal Experiments (JSAAE) skin irritation assay validation study management team (VMT). The 25 chemicals were listed in the European Centre for the Validation of Alternative Methods (ECVAM) performance standards. The reconstructed tissues were exposed to the chemicals for 15 minutes and incubated for 42 hours in fresh culture medium. Subsequently, the level of interleukin-1 alpha (IL-1α) present in the conditioned medium was measured, and tissue viability was assessed by using the MTT assay. The results of the MTT assay
obtained with the LabCyte EPI-MODEL24 (LabCyte MTT assay) demonstrated high within-laboratory and between-laboratory reproducibility, as well as high accuracy for use as a stand-alone assay to distinguish skin irritants from non-irritants. In addition, the IL-1α release measurements in the LabCyte assay were clearly unnecessary for the success of this model in the classification of chemicals for skin irritation potential.
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Ethical Issues in the Production of Human Cell Lines and Stem Cell Lines

Michael Balls

Response to question whether FRAME’s “alternative testing methods to animals on cosmetics include testing on stem cells from aborted babies/fetuses or any material whatsoever from aborted babies/fetuses”.
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The In Vitro Use of the Hair Follicle Closure Technique to Study the Follicular and Percutaneous Permeation of Topically Applied Drugs

Jessica Stahl, Frank Niedorf, Mareike Wohlert and Manfred Kietzmann

Recent studies on follicular permeation emphasise the importance of hair follicles as diffusion pathways, but only a limited amount of data are available about the follicular permeation of topically applied drugs. This study examines the use of a hair follicle closure technique in vitro, to determine the participation of hair follicles in transdermal drug penetration. Various substances, with different lipophilicities, were tested: caffeine, diclofenac, flufenamic acid, ibuprofen, paracetamol, salicylic acid and testosterone. Diffusion experiments were conducted with porcine skin, the most common replacement material for human skin, in Franz-type diffusion cells over 28 hours. Different experimental settings allowed the differentiation between interfollicular and follicular permeation after topical application of the test compounds. A comparison of the apparent permeability coefficients of the drugs demonstrates that the percutaneous permeations of caffeine and flufenamic acid were significantly higher along the hair follicles. In the cases of paracetamol and testosterone, the follicular pathway appears to be of importance, while no difference was found between interfollicular and follicular permeation for diclofenac, ibuprofen and salicylic acid. Thus, the hair follicle closure technique represents an adequate in vitro method for gaining information about follicular or percutaneous permeation, and can replace in vivo testing in animals or humans.
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