ALTEX最新文献

{"title":"Evaluation of EndoSens: A reliable skin sensitization assessment model.","authors":"Feiya Luo, Wenting Liao, Xinrong Pei, Shuxia Xing, Lei Sun, Lizhen Huang, Yong Lu","doi":"10.14573/altex.2602051","DOIUrl":"https://doi.org/10.14573/altex.2602051","url":null,"abstract":"<p><p>The transition toward non-animal safety assessment has driven the development of in vitro models for skin sensitization, particularly those targeting key event 2 (KE2) within the adverse outcome pathway (AOP). While conventional KE2 assays often rely on randomly integrated reporter systems, the EndoSens model utilizes CRISPR/Cas9-mediated knock-in to precisely insert a luciferase reporter into the endogenous HMOX1 locus in HaCaT keratinocytes, providing a genomically anchored and physiologically relevant system. To enable reproducible adoption across laboratories, we established a standardized protocol with defined quality control (QC) criteria, incorporating cinnamyl alcohol as a quantitative positive control and a cytotoxicity pre-screen to identify non-cytotoxic test concentrations. Evaluation using ten OECD TG 442D reference chemicals and ten additional compounds demonstrated the assay's robust performance in discriminating skin sensitizers with high inter-laboratory consistency, supporting its integration as a reliable in vitro component within defined approaches (DA) for animal-free skin sensitization assessment.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":""},"PeriodicalIF":5.8,"publicationDate":"2026-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147848879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Serum-free in vitro assessment of receptor-mediated endocrine activity including Phase-1 metabolism.","authors":"Inska Reichstein, Denise Horte, Rolf Marschalek, Eric Kowarz, Henner Hollert, Andreas Schiwy","doi":"10.14573/altex.2510171","DOIUrl":"https://doi.org/10.14573/altex.2510171","url":null,"abstract":"<p><p>Endocrine disruptors, which pose a risk to organisms and entire ecosystems even at low concentrations, can be detected by using standardized in vitro methods according to OECD test guidelines 455 and 458. However, these methods have disadvantages of (1) containing undefined animal supplements like fetal bovine serum (FBS) possibly leading to less reproducible results and ethical concerns and (2) lacking metabolic representation of whole organisms which could lead to an over- or underestimation of the samples' endocrine activity. In our study, we addressed both aspects by developing a simple, cost-effective FBS-free medium for culturing the OECD-relevant cell lines ERα-CALUX® and AR-CALUX®, and by integrating S9-homogenates (rat-S9, human S9 and their animal-free alternatives ewoS9R and ewoS9H) into OECD test procedures to simulate mammalian metabolism. We found that both OECD test procedures could be adapted to FBS-free conditions, yielding results similar to the FBS-containing assays, though some of the assessed model compounds were only metabolized in FBS-free or FBS-containing medium. Additionally, we found that rat and human S9 metabolized most of the compounds assessed, whereas ewoS9R and ewoS9H changed the endocrine activity of fewer substances. Maximum changes in the biologically equivalent concentrations of a 296-fold bioactivation and a 1,540-fold detoxification were observed, which can make a substantial difference for risk assessment. Overall, our study demonstrated that OECD 455 and 458 can be performed under FBS-free conditions and with S9-homogenates of different origins, enabling endocrine activity testing using less or no animal components while also enhancing the prediction to the in vivo situation.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":""},"PeriodicalIF":5.8,"publicationDate":"2026-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147731068","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tracing progress: The evolution of a protocol for relative metal release in surrogate gastric fluid.","authors":"Adriana R Oller, Pilar Prieto, Katherine Heim, João Barroso, Violaine Verougstraete","doi":"10.14573/altex.2512181","DOIUrl":"https://doi.org/10.14573/altex.2512181","url":null,"abstract":"<p><p>Grouping and read-across are applied to human health toxicity classification of metal substances under EU REACH to decrease animal testing. Metal ion release is responsible for the systemic (and often local) toxicity of metal-containing materials. Metal release in simulated gastric fluid is relevant to the oral route of exposure. In 2010, the metals' industry initiated the development of a method to assess relative metal release in HCl pH 1.5 from different substances of the same metal, including metal-containing materials such as massive and powder forms of alloys, metals, inorganic metal compounds, and other inorganic complex metal-containing materials. These data could also identify matrix effects in complex materials that increase or decrease metal release compared to what could be predicted from the content of the classified ingredients. The method underwent a round robin and multiple reviews over the past 15 years (e.g. by EURL ECVAM, ESAC and OECD), leading to revisions of the protocol. This paper provides a history of the protocol's evolution, and details supporting studies and revisions made to address the comments and concerns of its reviewers. The 2025 version of the protocol (publicly available via TSAR) significantly improved its clarity and the robustness of the method without changing the way in which data are generated. The method (and associated 2025 protocol) is relevant to the oral route and is suitable for supporting grouping and read-across of metal-containing materials, and refining the classification of metal materials showing a matrix effect, which might contribute to reducing animal testing.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":""},"PeriodicalIF":5.8,"publicationDate":"2026-04-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147731036","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Stakeholder input towards further refinement and consolidation of the alternative safety profiling algorithm (ASPA) for next generation risk assessment (NGRA).","authors":"Mirjam Luijten, Matthias Herzler, Femke Affourtit, Dave Allen, Muhammad Waqar Ashraf, Nicholas Ball, Elisabet Berggren, Sandra Berndt, Pierre-André Billat, Eike Cöllen, John Colbourne, Marco Marco, Richard Currie, Michael Guy Diemar, Hubert Dirven, Nadine Dreser, Sylvia E Escher, Iain Gardner, Rafael Gozalbes, Stefan Hahn, Astrid Heiland, Claudia Hempt, Giel Hendriks, Oliver Henschel, Trine Husøy, Barira Islam, Keld Alstrup Jensen, Hennicke Kamp, Spyros Karakitsios, George E N Kass, Vikas Kumar, Robert Landsiedel, Thomas Loret, Patrik Lundquist, Catherine Mahony, Philip Marx-Stoelting, Martijn J Moné, Ilse Ottenbros, Gladys Ouédraogo, Manuel Pastor, Magdalini Sachana, Sebastian Schmeisser, Stefan Scholz, Katrin Schütte, Kristie Sullivan, Silvia Tangianu, Adrian Tristram, Janette Turner, Mathieu Vinken, Jelle Vlaanderen, Bob van de Water, Andrew White, Marcel Leist","doi":"10.14573/altex.2603121","DOIUrl":"https://doi.org/10.14573/altex.2603121","url":null,"abstract":"<p><p>Next-generation risk assessment (NGRA) aims to enable transparent, reproducible chemical safety assessments based on human-relevant, animal-free new approach methodologies (NAMs). The Alternative Safety Profiling Algorithm (ASPA) was developed within the ASPIS cluster to provide an algorithmic workflow that structures problem formulation, evidence integration, and decision-making across three main pillars-hazard, ADME (toxicokinetics), and exposure. To refine ASPA, a stakeholder workshop was organized. Four breakout groups systematically reviewed corresponding workflow sections, identifying strengths, conceptual gaps, and opportunities for harmonization. Across groups, participants endorsed ASPA's modular, technology-neutral nature and its focus on standardizing processes rather than prescribing specific test batteries. The hazard pillar discussions emphasized a sensitive, hypothesis-generating Tier 1, complemented by a specific, mechanistic Tier 2, capable of deriving points of departure (PoDs). ADME experts supported a physiology-based kinetic (PBK) modelling strategy, advancing from generic towards more complex models, using mechanistic information and experimental data. The exposure group proposed refinements for transparent, tiered exposure modelling, with emphasis on realistic worst-case scenarios and explicit uncertainty communication. Cross-pillar discussions highlighted the importance of feedback loops between all pillars, and the documentation of decision points to achieve consistency and defensibility. The workshop outcomes informed three parallel developments: (i) algorithmic refinement and re-design toward the next ASPA version, (ii) the creation of detailed guidance for each building block, and (iii) the establishment of practical case studies to demonstrate workflow implementation. This report already contains a first case study (developmental neurotoxicity assessment of desnitro-imidacloprid). These advances increase the operability, transparency, and regulatory readiness of ASPA.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":""},"PeriodicalIF":5.8,"publicationDate":"2026-04-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147701826","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Investigation of the potential of bisphenol A substitutes to induce allergic contact sensitization using OECD defined approaches.","authors":"Victor J Johnson, Travis V Gulledge, Michael I Luster, Gary R Burleson, Florence G Burleson, Dori Germolec","doi":"10.14573/altex.2512011","DOIUrl":"https://doi.org/10.14573/altex.2512011","url":null,"abstract":"<p><p>Bisphenol A is a high production volume chemical used extensively in the manufacture of polycarbonate plastics, epoxy resins, and thermal printer paper with a high potential for occupational and post-production dermal exposure. Bisphenol A-containing plastics were commonly used in food packaging resulting in significant public exposure through leaching into foodstuff. The public is also at risk of dermal exposure due to environmental contamination. Due to public health concerns regarding the potential for endocrine disrupting effects, efforts have been applied to replace bisphenol A with safer alternatives. Bisphenol A has been shown to cause skin sensitization in humans; however, there is a paucity of information available on the sensitizing potential of structural analogues which are increasingly being employed as substitutes. We utilized new approach methodologies (NAMs) addressing key events 1-3 of the adverse outcome pathway for skin sensitization to address the potential of bisphenol A substitutes to induce dermal sensitization. Defined approaches (DA) were applied to further classify and categorize potency according to OECD TG 497. The NAMs and DAs confirmed that bisphenol A was a skin sensitizer in potency category UN GHS 1B. Bisphenol B, AP, and E were also classified as UN GHS 1B sensitizers, and bisphenol AF as UN GHS 1A/1B depending on DA, while 2,4-bisphenol S and F were borderline sensitizers, and bisphenol S was classified as a non-sensitizer. These data provide evidence of skin sensitization hazard for the bisphenol structural analogues tested, except for bisphenol S, suggesting that they present risks for dermal allergy.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":""},"PeriodicalIF":5.8,"publicationDate":"2026-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147663524","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Implementation of reduction and refinement-related parameters in repeated dose toxicity studies.","authors":"Katy Taylor, Laura Rego Alvarez, Emma Grange, David Andrew","doi":"10.14573/altex.2510071","DOIUrl":"https://doi.org/10.14573/altex.2510071","url":null,"abstract":"<p><p>The OECD's Test Guideline programme provides internationally recognised standard methods for non-clinical health and environmental safety testing of chemicals. They act as a harmonisation mechanism and can support the replacement, reduction and refinement of regulatory tests using animals. We reviewed the reports of 300 repeated dose tests using rats conducted for chemical safety purposes to determine the extent to which current reduction- and refinement-related parameters in TG 407 and 408 had been taken up. We found strong adherence to minimum group size and number of dose groups and strong adherence to the maximum dose level and dose volume. There was, however, very low uptake of the use of a limit test as a reduction method in both TGs, and some evidence of continued use of individual housing and larger than the minimum group sizes in the 28-day studies (TG 407). There are other opportunities to improve the TGs with respect to reduction and refinement, and we recommend that there is a systematic review by the OECD test guideline programme to ensure their guidelines comply with the aims of the 3Rs.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":""},"PeriodicalIF":5.8,"publicationDate":"2026-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147701814","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Validation study for the evaluation of MCTT HCE™ test method for eye safety assessment of medical devices.","authors":"Nam-Hee Kang, Seolyeong Kim, Subin Lee, Jungeun Park, Mi-Sook Jung, Hyo-Ju An, Min-Gyu Go, Joohwan Kim, Sun Wook Woo","doi":"10.14573/altex.2511041","DOIUrl":"https://doi.org/10.14573/altex.2511041","url":null,"abstract":"<p><p>According to ISO 10993-23 (Biological evaluation of medical devices - Part 23: Tests for irritation, Annex D2), no specific alternative test method has yet been established to identify substances that may cause eye irritation or serious eye damage in medical devices. Therefore, the in vivo animal test OECD TG 405 continues to be used. This study adapted the in vitro eye irritation test (EIT) using the MCTT HCE™ Reconstructed Human Corneal Epithelial (RHCE) model, which was originally developed for testing chemicals, to the testing of ophthalmic medical devices such as contact lenses and intraocular lenses. A validation study, carried out in accordance with OECD GD 34, aimed to confirm transferability (2 produced medical devices and 2 spiked extract vehicles), proficiency (4 produced medical devices and 3 spiked extract vehicles), and reproducibility (8 produced medical devices and 9 spiked extract vehicles). The results demonstrated a combined predictive capacity of 89.6% sensitivity, 87.3% specificity, and 88.4% accuracy based on 285 runs. Specifically, the test showed 85.7% sensitivity, 100% specificity, and 92.9% accuracy for classifying produced medical devices (140 runs) and 93.8% sensitivity, 76.3% specificity, and 84.1% accuracy for classifying solutions spiked with chemicals before extraction (145 runs). These findings confirm the reliability of the 'MCTT HCE™ EIT for medical devices' in assessing the irritation potential of medical device extracts, including the detection of low concentrations of potent irritants.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":""},"PeriodicalIF":5.8,"publicationDate":"2026-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147577543","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Multiparametric evaluation of different FBS-free replacement media for widely used human cancer cell lines.","authors":"Anne L Koelz, Claudia Pommerenke, Pascal Woitschewski, Yvonne Merkhoffer, Wilhelm G Dirks, Sonja Eberth","doi":"10.14573/altex.2512111","DOIUrl":"https://doi.org/10.14573/altex.2512111","url":null,"abstract":"<p><p>Despite ethical concerns and scientific drawbacks, fetal bovine serum (FBS) remains a common supplement of culture media for continuous cancer cell lines. Although FBS alternatives like human platelet lysate (hPL) and animal component-free, chemically defined media (CDM) are commercially available for many years, acceptance of alternative media is limited, as data verifying the stability of the phenotype and function of a cell line in the alternatives are often lacking. Here, we have adapted four widely used human cancer cell lines (HELA, HL-60, JIMT-1, K-562) to hPL supplemented media and different CDM. To evaluate the FBS-free replacements in comparison to FBS media, we systematically analyzed the cultures in respect to recovery after cryopreservation, short tandem repeat (STR) profile stability, proliferation, morphology and transcriptomic alterations. Neither changes in STR profiles nor difficulties after cryopreservation were detected. With the exception of K-562, FBS-free cultures showed a reduced proliferation rate and, in some conditions, slight morphological alterations in comparison to FBS cultures. In all cell lines, gene set enrichment analyses revealed that culture media mainly affected expression of cholesterol homeostasis genes. In HELA and JIMT-1, media also influenced genes of epithelial-mesenchymal transition, nevertheless their overall phenotypic hallmarks remained stable. Only a few differentiation markers were among the differentially expressed genes of HL-60 and K-562 cultures, while their main phenotypes remained unchanged. This was further confirmed by successful induction of differentiation of HL-60 in FBS-free media. In conclusion, our multiparametric approach provides strong evidence supporting the transition to FBS-free media for even long-established cancer models.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":" ","pages":""},"PeriodicalIF":5.8,"publicationDate":"2026-03-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147518217","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pyrogen testing at a turning point - On occasion of the 30th anniversary of the whole blood monocyte activation test.","authors":"Thomas Hartung","doi":"10.14573/altex.2603201","DOIUrl":"https://doi.org/10.14573/altex.2603201","url":null,"abstract":"<p><p>The whole blood pyrogen test was first described in this journal exactly thirty years ago. Its variant based on cryopreserved blood followed one year later. Together with other monocyte activation tests (MATs), it has fundamentally changed the landscape of pyrogen testing. In the five years since the 25th anniversary article in this series, progress has been remarkable: The European Pharmaco-poeia deleted the rabbit pyrogen test (RPT) effective January 2026, ending a 55-year era; ISO 10993-1:2025 removed material-mediated pyrogenicity from mandatory evaluation endpoints for medical devices; the U.S. FDA updated its guidance on pyrogen testing; and the MAT market grew to over $600 million. New validation studies have demonstrated MAT equivalence to the RPT for both endotoxin and non-endotoxin pyrogens, and the first product-specific MAT validations have been accepted in regulatory filings in Europe and the United States. Reporter cell lines and tran-scriptomic approaches are opening next-generation detection capabilities. Yet implementation gaps persist: The MAT is still underutilized for blood transfusions, cell therapies, and airborne pyrogens. Recombinant alternatives to the Limulus amebocyte lysate assay (LAL) have finally achieved phar-macopeial recognition, addressing horseshoe crab conservation concerns. This article reviews the developments of the last five years, updates the lessons learned, and reflects on three decades of bringing a human cell-based test from the laboratory bench to global regulatory acceptance.</p>","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":"43 2","pages":"199-214"},"PeriodicalIF":5.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147694640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advancing humane and human-relevant science: The 8th Annual Conference of the Society for Alternatives to Animal Experiments - India (SAAE-I 2025).","authors":"Bokara Kiran Kumar, Mohammad A Akbarsha, Kasturi Mahadik, Mohammed M Idris","doi":"10.14573/altex.2511291","DOIUrl":"10.14573/altex.2511291","url":null,"abstract":"","PeriodicalId":520550,"journal":{"name":"ALTEX","volume":"43 2","pages":"359-362"},"PeriodicalIF":5.8,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147694616","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}