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MatTek Corporation has been working diligently for over 15 years to replace traditional animal-based toxicity and efficacy tests with alternative test methods based on human-cell derived, three-dimensional (3-D) tissue models. First discussed in detail by W.M.S. Russell and R.L. Burch 50 years ago in their book, The Principles of Humane Experimenta...
Citations
... However, the model is well-established for testing to radiation toxicity and alterations. Furthermore, applying skin models for radiotherapy experiments is also an option for avoiding/reducing the use of animal models [52]. ...
Background:
Inflammatory skin reactions and skin alterations are still a potential side effect in radiation therapy (RT), which also need attention for patients' health care.
Method:
In a pre-clinical study we consider alterations in irradiated in-vitro skin models of epidermal and dermal layers. Typical dose regimes in radiation therapy are applied for irradiation. For non-invasive imaging and characterization optical coherence tomography (OCT) is used. Histological staining method is additionally applied for comparison and discussion.
Results:
Structural features, such as keratinization, modifications in epidermal cell layer thickness and disorder in the layering-as indications for reactions to ionizing radiation and aging-could be observed by means of OCT and confirmed by histology. We were able to recognize known RT induced changes such as hyper-keratosis, acantholysis, and epidermal hyperplasia as well as disruption and/or demarcation of the dermo-epidermal junction.
Conclusion:
The results may pave the way for OCT to be considered as a possible adjunctive tool to detect and monitor early skin inflammation and side effects of radiotherapy, thus supporting patient healthcare in the future.
... 8 Fortunately, three-dimensional, organotypic tissue explants of human upper respiratory mucosa are now available. [9][10][11] These models more reliably mimic the function of living tissue than either animal models or cell culture monolayers and can greatly accelerate drug discovery. 9,11 Using these models, we were thus able to study the biochemical pathway of pharyngitis in great detail and design a scientifically based treatment. ...
Background
Often thought of as a minor health concern, sore throat or pharyngitis is an important public health issue. It is one of the most common symptoms of upper respiratory diseases including COVID-19 and is a leading cause of physician visits and antibiotic prescriptions. However, few over-the-counter medications are proven to heal sore throat inflammation.
Methods
Adenocarcinomic human alveolar basal epithelial cells (A549 cells) and three dimensional organotypic human respiratory tissues were used to study inflammation and various treatment effects on respiratory epithelia. The cells and tissues were studied both in the presence and absence of bradykinin, one of the first inflammatory mediators of pharyngitis. Inflammation was measured by analyzing the levels of prostaglandin E2 (PGE2), interleukin 8 (IL-8), and leukotriene B4 (LTB4), transepithelial electrical resistance (TEER), and lactate dehydrogenase (LDH) release. Tissue morphology was analyzed by immunohistochemistry.
Results
In studying pharyngitis using organotypic human respiratory tissue stimulated with bradykinin, we saw an increase in PGE2 and interleukin-8 (IL-8) in response to bradykinin. Acetyl salicylic acid (ASA), a nonspecific COX inhibitor, was able to mitigate a bradykinin-induced increase in PGE2 in our studies. However, ASA was inflammatory above its therapeutic window, increasing the levels of PGE2 and IL-8 above those seen with bradykinin stimulation alone. We describe a novel, scientifically validated treatment for sore throat, that contains a low dose of ASA and other anti-inflammatory ingredients.
Conclusion
This study elucidates the complex mechanisms involved in healing pharyngitis, an inflammatory condition of the upper respiratory epithelia. An ASA-based formula (Biovanta) mitigated bradykinin-induced inflammation more strongly than ASA alone in organotypic human respiratory tissues. Surprisingly, we found that many of the most common over the counter sore throat therapies exacerbate inflammation and IL-8 in organotypic human respiratory tissues, suggesting these common treatments may increase the likelihood of further respiratory complications.
... The needs of the cosmetic industry and the regulatory agencies for in vitro tissue analogs to replace animal testing for eye irritation tests continue to grow (Gruber and Hartung 2004;Barile 2010;Abdelkader et al. 2015;Ghezzi et al. 2015). Since 3D models allow direct topical application of the undiluted test materials onto the tissue surface, the models have been used widely by the cosmetic industry to evaluate the irritation potential of raw materials, compound mixtures, and surfactant-based formulations that are designed for use in the vicinity of the eye (Stern et al. 1998;Jones et al. 2001;Sheasgreen et al. 2009;Jirova et al. 2014). The 3D models are used by contract research laboratories, cosmetic, personal care, and household chemical companies in place of Draize testing for product development. ...
Testing of all manufactured products and their ingredients for eye irritation is a regulatory requirement. In the last two decades, the development of alternatives to the in vivo Draize eye irritation test method has substantially advanced due to the improvements in primary cell isolation, cell culture techniques, and media, which have led to improved in vitro corneal tissue models and test methods. Most in vitro models for ocular toxicology attempt to reproduce the corneal epithelial tissue which consists of 4–5 layers of non-keratinized corneal epithelial cells that form tight junctions, thereby limiting the penetration of chemicals, xenobiotics, and pharmaceuticals. Also, significant efforts have been directed toward the development of more complex three-dimensional (3D) equivalents to study wound healing, drug permeation, and bioavailability. This review focuses on in vitro reconstructed 3D corneal tissue models and their utilization in ocular toxicology as well as their application to pharmacology and ophthalmic research. Current human 3D corneal epithelial cell culture models have replaced in vivo animal eye irritation tests for many applications, and substantial validation efforts are in progress to verify and approve alternative eye irritation tests for widespread use. The validation of drug absorption models and further development of models and test methods for many ophthalmic and ocular disease applications is required.
... Since these medications have come to market, scientific research techniques in the field have greatly advanced. Although there are still arguably very few reliable animal models to study upper respiratory disease, three-dimensional, organotypic tissue explants of upper respiratory mucosa are now available (4)(5)(6). These models more reliably mimic the function of living tissue than either animal models or cell culture monolayers and can greatly accelerate drug discovery (4,6). ...
... Mattek Corporation (Ashland MA) (5,7). The tissue we use in our studies are generated either from nasal biopsy or trachea-bronchial tissue. ...
Often thought of as a minor health concern, sore throat or pharyngitis is an important public health issue. It is one of the most common symptoms of upper respiratory diseases including COVID-19 and is a leading cause of physician visits and antibiotic prescriptions. Despite being on the market for decades, few over the counter sore throat medications are proven to heal sore throat. In studying pharyngitis using organotypic human respiratory tissue stimulated with bradykinin, we saw an increase in prostaglandin E2 (PGE2) and interleukin-8 (IL-8) in response to bradykinin. Bradykinin is one of the first inflammatory signals for pharyngitis and it increases PGE2 in human subjects. If left unregulated, PGE2 may further increase inflammation via the COX pathway and via IL-8, a proinflammatory chemokine responsible for neutrophil infiltration and possibly thus, a cytokine storm. Acetyl salicylic acid (ASA), a non-specific COX inhibitor, was able to mitigate a bradykinin-induced increase in PGE2 in our studies. However, ASA was inflammatory above its small therapeutic window, greatly increasing levels of PGE2 and IL-8 above those seen with bradykinin stimulation alone. Similar to other systems, the respiratory epithelia maintains a delicate balance of pro-inflammatory and anti-inflammatory signals in order to keep the respiratory barrier intact. To our knowledge, this is the first study to try and elucidate the complex mechanisms involved in healing pharyngitis, an inflammatory condition of the respiratory epithelia. Biovanta™, a formula containing ASA mitigated bradykinin-induced inflammation more strongly than ASA alone in organotypic human respiratory tissues. Surprisingly, we found that many of the most common over the counter sore throat therapies exacerbate inflammation and IL-8 in organotypic human respiratory tissues, suggesting these common treatments may possibly increase the likelihood of further respiratory complications in people.
Competing interest statement
This study was funded entirely by Applied Biological Laboratories, a private company that owns the Biovanta™ product. Unless otherwise indicated all experiments were performed at Applied Biological Laboratories research facility located at the SUNY Downstate Biotechnology Incubator, a part of StartUP NY. All of the authors were employees of Applied Biological Laboratories at the time the experiments were performed.
... In addition, researchers must be aware of the local and international guidelines that regulate ethical experimental conduct with animals, and, whenever possible, the application of the principle of the 3R's. Advances in the science have resulted in the creation of several in vitro models (e.g., micro-chips with organs, 3D tissue models, blood derivatives, computer modelling) (Huh et al., 2013;Sheasgreen et al 2009;Sladowski et al., 2001;Martonen et al., 2003;Aguda et al., 2011) potentially replacing and reducing animal experimentation. Researchers are encouraged to incorporate alternative models when experimentally feasible. ...
Certain chronic conditions like HIV, diabetes and hypertension may lead to neurocognitive impairment (NCI) (Heaton et al., 2010; Iadecola et al., 2016; Zilliox, Chadrasekaran, Kwan, & Russell, 2016). These frequently co-occurring conditions can, by implication, affect vocational functioning. There is a need to understand how such impairment may affect society vis-à-vis the work place, particularly as the relationship between neurocognitive performance and real-world vocational functioning is a determinant of employability and job placement, and it affects safety and medical treatment options. As the remarkable effectiveness of modern medicine allows increasing numbers of people living with HIV (PLWH) to return to or remain in the workplace, it is becoming paramount to effectively manage HIV-associated neurocognitive disorders (HAND) in high risk workplace
settings, such as driving. Studies are needed to develop ways to identify impaired drivers while at the same time protecting the public and the individual. Working within existing research ethics frameworks (Belmont Report (Office for Human Research Protections, 2016) and Declaration of Helsinki (World Health Organization, 2001) we use our project as a case study to discuss ethical challenges associated with conducting research in professional drivers living with HIV in South Africa.
... Using Russel and Burch's principle of replacement (Russell and Burch 1959), several human-derived three-dimensional models have been synthesised, tested, validated (a few of the several), and used (Sheasgreen et al. 2009). These 3-D in vitro models have not only an "ethical edge", but also a more pathophysiologically relevant edge owing to the human origin of these tissues. ...
... Additionally, an animal model (like a commonly used inbred murine model) has far less biological and genetic differences compared to the complex human genetic/biological heterogeneity, making human tissue much more physiologically relevant. These three-dimensional human-derived tissue models include oral epithelia (Klausner et al. 2007), gastrointestinal epithelia (Sheasgreen et al. 2009), vaginal epithelia (Ayehunie et al. 2011), ocular tissue (Kaluzhny et al., 2011), gingival tissue (Hai et al. 2006), respiratory epithelia (Sexton et al. 2011), and dendritic antigen-presenting cells (Sheasgreen et al. 2009). ...
... Additionally, an animal model (like a commonly used inbred murine model) has far less biological and genetic differences compared to the complex human genetic/biological heterogeneity, making human tissue much more physiologically relevant. These three-dimensional human-derived tissue models include oral epithelia (Klausner et al. 2007), gastrointestinal epithelia (Sheasgreen et al. 2009), vaginal epithelia (Ayehunie et al. 2011), ocular tissue (Kaluzhny et al., 2011), gingival tissue (Hai et al. 2006), respiratory epithelia (Sexton et al. 2011), and dendritic antigen-presenting cells (Sheasgreen et al. 2009). ...
Animals have been used in research and teaching for a long time. However, clear ethical guidelines and pertinent legislation were instated only in the past few decades, even in developed countries with Judeo-Christian ethical roots. We compactly cover the basics of animal research ethics, ethical reviewing and compliance guidelines for animal experimentation across the developed world, “our” fundamentals of institutional animal research ethics teaching, and emerging alternatives to animal research. This treatise was meticulously constructed for scientists interested/involved in animal research. Herein, we discuss key animal ethics principles – Replacement/Reduction/Refinement. Despite similar undergirding principles across developed countries, ethical reviewing and compliance guidelines for animal experimentation vary. The chronology and evolution of mandatory institutional ethical reviewing of animal experimentation (in its pioneering nations) are summarised. This is followed by a concise rendition of the fundamentals of teaching animal research ethics in institutions. With the advent of newer methodologies in human cell-culturing, novel/emerging methods aim to minimise, if not avoid the usage of animals in experimentation. Relevant to this, we discuss key extant/emerging alternatives to animal use in research; including organs on chips, human-derived three-dimensional tissue models, human blood derivates, microdosing, and computer modelling of various hues.
... MatTek Corporation developed a commercially available 3D corneal epithelial model with primary human epidermal keratinocytes (62)(63)(64). The keratinocytes are grown on cell-culture inserts in serum-free media, to form a stratified, squamous epithelium, EpiOcular TM . ...
Human eyes and skin are frequently exposed to chemicals accidentally or on purpose due to their external location. Therefore, chemicals are required to undergo the evaluation of the ocular and dermal irritancy for their safe handling and use before release into the market. Draize rabbit eye and skin irritation test developed in 1944, has been a gold standard test which was enlisted as OECD TG 404 and OECD TG 405 but it has been criticized with respect to animal welfare due to invasive and cruel procedure. To replace it, diverse alternatives have been developed: (i) For Draize eye irritation test, organotypic assay, in vitro cytotoxicity-based method, in chemico tests, in silico prediction model, and 3D reconstructed human cornea-like epithelium (RhCE); (ii) For Draize skin irritation test, in vitro cytotoxicity-based cell model, and 3D reconstructed human epidermis models (RhE). Of these, RhCE and RhE models are getting spotlight as a promising alternative with a wide applicability domain covering cosmetics and personal care products. In this review, we overviewed the current alternatives to Draize test with a focus on 3D human epithelium models to provide an insight into advancing and widening their utility.
... The MatTek Corporation developed a commercially available 3D corneal epithelial model (OCL-200) based upon human derived epidermal keratinocytes from neonatal human foreskin (McLaughlin et al., 2009;Sheasgreen et al., 2009) grown on cell-culture inserts in serum-free media, to form a stratified, squamous epithelium, marketed as EpiOcular TM . Test substances are directly applied to the models, and cytotoxicity is measured using MTT. ...
... A test cannot proceed to peer review without a VMG recommendation. A formal regulatory validation can take more than five years to achieve (Sheasgreen et al., 2009) and may only then be considered for regulatory acceptance once achieved. ...
Given the hazardous nature of many materials and substances, ocular toxicity testing is required to evaluate the dangers associated with these substances after their exposure to the eye. Historically, animal tests such as the Draize test were exclusively used to determine the level of ocular toxicity by applying a test substance to a live rabbit’s eye and evaluating the biological response. In recent years, legislation in many developed countries has been introduced to try to reduce animal testing and promote alternative techniques. These techniques include ex vivo tests on deceased animal tissue, computational models that use algorithms to apply existing data to new chemicals and in vitro assays based on two dimensional (2D) and three dimensional (3D) cell culture models. Here we provide a comprehensive overview of the latest advances in ocular toxicity testing techniques, and discuss the regulatory framework used to evaluate their suitability.
... The MatTek Corporation has developed a commercially available 3D corneal epithelial model based upon human derived epidermal keratinocytes from human foreskin [28,255], marketed as EpiOcular TM . Although it is been used by numerous cosmetic companies in place of Draize testing, as of yet EpiOcular TM has not been formally validated [255] as it is unable to predict responses of chemicals that affect the lower layers of the cornea, or that are dependent upon epithelial-stromal cellular interactions [28]. ...
... The MatTek Corporation has developed a commercially available 3D corneal epithelial model based upon human derived epidermal keratinocytes from human foreskin [28,255], marketed as EpiOcular TM . Although it is been used by numerous cosmetic companies in place of Draize testing, as of yet EpiOcular TM has not been formally validated [255] as it is unable to predict responses of chemicals that affect the lower layers of the cornea, or that are dependent upon epithelial-stromal cellular interactions [28]. ...
The worldwide limited availability of suitable corneal donor tissue has led to the development of alternatives, including keratoprostheses (Kpros) and tissue engineered (TE) constructs. Despite advances in bioscaffold design, there is yet to be a corneal equivalent that effectively mimics both the native tissue ultrastructure and biomechanical properties. Human decellularized corneas (DCs) could offer a safe, sustainable source of corneal tissue, increasing the donor pool and potentially reducing the risk of immune rejection after corneal graft surgery. Appropriate, human-specific, decellularization techniques and high-resolution, non-destructive analysis systems are required to ensure reproducible outputs can be achieved. If robust treatment and characterization processes can be developed, DCs could offer a supplement to the donor corneal pool, alongside superior cell culture systems for pharmacology, toxicology and drug discovery studies.
... Therefore, the EpiOcular TM eye irritation assay [6], was used to determine the relative chemical irritation produced by lunar dust prior to in vivo testing. This human-derived in vitro model has been used worldwide by chemical, pharmaceutical and consumer product companies in testing approaches to reduce or replace animal testing [10,11]. Results reported by Stillmeadow, Inc. are shown in Table 1. ...
Background
Dust exposure is a well-known occupational hazard for terrestrial workers and astronauts alike and will continue to be a concern as humankind pursues exploration and habitation of objects beyond Earth. Humankind’s limited exploration experience with the Apollo Program indicates that exposure to dust will be unavoidable. Therefore, NASA must assess potential toxicity and recommend appropriate mitigation measures to ensure that explorers are adequately protected. Visual acuity is critical during exploration activities and operations aboard spacecraft. Therefore, the present research was performed to ascertain the ocular toxicity of authentic lunar dust.
Methods
Small (mean particle diameter = 2.9 ± 1.0 μm), reactive lunar dust particles were produced by grinding bulk dust under ultrapure nitrogen conditions. Chemical reactivity and cytotoxicity testing were performed using the commercially available EpiOcularTM assay. Subsequent in vivo Draize testing utilized a larger size fraction of unground lunar dust that is more relevant to ocular exposures (particles <120 μm; median particle diameter = 50.9 ± 19.8 μm).
Results
In vitro testing indicated minimal irritancy potential based on the time required to reduce cell viability by 50% (ET50). Follow-up testing using the Draize standard protocol confirmed that the lunar dust was minimally irritating. Minor irritation of the upper eyelids was noted at the 1-hour observation point, but these effects resolved within 24 hours. In addition, no corneal scratching was observed using fluorescein stain.
Conclusions
Low-titanium mare lunar dust is minimally irritating to the eyes and is considered a nuisance dust for ocular exposure. No special precautions are recommended to protect against ocular exposures, but fully shielded goggles may be used if dust becomes a nuisance.
