In vitro quantitative determination of ophthalmic irritancy by the chorioallantoic membrane test with trypan blue staining as alternative to eye irritation test

Lagarto, Alicia; Vega, Roser; Guerra, Idania Rodeiro; González, Rafael de Miguel

doi:10.1016/j.tiv.2005.10.003

Cited by 23 publications

(6 citation statements)

References 11 publications

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“…of Shiseido Co, developed an objective evaluation tool for the CAM assay by employing trypan blue staining (CAM-TB) (33). The CAM-TB assay adopts trypan blue uptake to indicate chorioallantoic membrane injury, which showed a good correlation with the in vivo eye test (30,33–35). However, since fertilized eggs eventually become chicken, HET-CAM test practically requires the sacrifice of animals.…”

Section: Alternatives To Draize Rabbit Eye Irritation Testmentioning

confidence: 97%

Alternatives toIn VivoDraize Rabbit Eye and Skin Irritation Tests with a Focus on 3D Reconstructed Human Cornea-Like Epithelium and Epidermis Models

2017

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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.

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Section: Alternatives To Draize Rabbit Eye Irritation Testmentioning

confidence: 97%

Alternatives toIn VivoDraize Rabbit Eye and Skin Irritation Tests with a Focus on 3D Reconstructed Human Cornea-Like Epithelium and Epidermis Models

2017

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“…It has also been expanded to include the chorio-allantoic membrane (CAM) test as a mucous membrane irritation test. 24 Only fresh fertile eggs weighing 50-60 g can be used in this test. The eggs were placed in an automatically rotating incubator and kept at a temperature of 37.5°C ± 0.5°C and relative humidity of 62.5% ± 7.5%.…”

Section: Enzymatic Hydrolysis Assaymentioning

confidence: 99%

In vivo and in vitro evaluation of octyl methoxycinnamate liposomes

Mota¹,

Freitas²,

Júnior³

et al. 2013

IJN

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Solar radiation causes damage to human skin, and photoprotection is the main way to prevent these harmful effects. The development of sunscreen formulations containing nanosystems is of great interest in the pharmaceutical and cosmetic industries because of the many potential benefits. This study aimed to develop and evaluate an octyl methoxycinnamate (OMC) liposomal nanosystem (liposome/OMC) to obtain a sunscreen formulation with improved safety and efficacy by retaining OMC for longer on the stratum corneum. Methods: The liposome/OMC nanostructure obtained was tested for enzymatic hydrolysis with lipase from Rhizomucor miehei and biodistribution with liposomes labeled with technetium-99m. The liposome/OMC formulation was then incorporated in a gel formulation and tested for ocular irritation using the hen's egg test-chorio-allantoic membrane (HET-CAM) assay, in vitro and in vivo sun protection factor, in vitro release profile, skin biometrics, and in vivo tape stripping. Results: The liposome/OMC nanosystem was not hydrolyzed from R. miehei by lipase. In the biodistribution assay, the liposome/OMC formulation labeled with technetium-99m had mainly deposited in the skin, while for OMC the main organ was the liver, showing that the liposome had higher affinity for the skin than OMC. The liposome/OMC formulation was classified as nonirritating in the HET-CAM test, indicating good histocompatibility. The formulation containing liposome/OMC had a higher in vivo solar photoprotection factor, but did not show increased water resistance. Inclusion in liposomes was able to slow down the release of OMC from the formulation, with a lower steady-state flux (3.9 ± 0.33 µg/cm 2 /hour) compared with the conventional formulation (6.3 ± 1.21 µg/cm 2 /hour). The stripping method showed increased uptake of OMC in the stratum corneum, giving an amount of 22.64 ± 7.55 µg/cm 2 of OMC, which was higher than the amount found for the conventional formulation (14.57 ± 2.30 µg/cm 2 ). Conclusion: These results indicate that liposomes are superior carriers for OMC, and confer greater safety and efficacy to sunscreen formulations.

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“…It has also been expanded to include the CAM test as a mucous-membrane irritation test. 33 On this test, only fresh, fertile eggs that weigh minimally 50 g and maximally 60 g can be used. They are put into an incubator, which automatically rotates and is maintained at an optimum temperature of 37.5 ± 0.5 C and a relative humidity of 62.5 ± 7.5%.…”

Section: Ophthalmic Irritancy By the Het-cam Testmentioning

confidence: 99%

“…The CAM-TBS test uses trypan blue absorption as an indicator of CAM injury and shows a good correlation with the in vivo Draize eye irritation test. 33 The procedure for this test was almost the same as the previous one, but in this case, a silicone ring with an 18-mm diameter was added to guarantee the accuracy of the test. After the substance was removed, 500 L of phosphate saline buffer (PBS) and 0.1% of TBS were put onto the CAM in the area limited by the silicone ring.…”

Section: Ophthalmic Irritancy By the Cam-tbs Testmentioning

confidence: 99%

“…The absorbance was measured spectrophotometrically at 595 nm by using the supernatant. 33 For the control, 4 eggs were used and followed the same procedure mentioned before, except for the fact that no substance was added to be analyzed. Each formulation was classified according to the mean value found with the analysis of the 4 eggs.…”

Section: Ophthalmic Irritancy By the Cam-tbs Testmentioning

confidence: 99%

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Characterization and Evaluation of Poly(ε-caprolactone) Nanoparticles Containing 2-Ethylhexyl-p-Methoxycinnamate, Octocrylene, and Benzophenone-3 in Anti-Solar Preparations

Nascimento

Silva²,

Mansur³

et al. 2012

j nanosci nanotechnol

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Ultraviolet radiation can bring both harm and benefits to human health. Among those harms are erythemas, photosensitivity, photoaging, and the most worrying, skin cancer. Nanoencapsulation of sunscreen agents (SA) by using a biocompatible and biodegradable polymer such as poly(epsilon-caprolactone) (PCL) is advantageous as it increases the retention of UV absorbers in the skin, avoids systemic absorption, and consequently, improves water resistance and stability of the preparation. The aim of this work is to develop, characterize, and study the encapsulation of 3 different SA: 2-ethylhexyl-p-methoxycinnamate, benzophenone-3, and octocrylene in PCL nanoparticles (Nps). Nps were prepared by the solvent emulsification and evaporation method. The process yield was calculated, and the Nps were characterized in terms of size, polydispersity index (PI), morphology, zeta potential (ZP), encapsulation efficiency (EE) (%), and sunscreen agent content (SAC). The final formulations were submitted to the hen's egg test-chorioallantoic membrane (HET-CAM), chorioallantoic membrane-trypan blue staining (CAM-TBS), red blood cell (RBC), Draize tests, in vitro release, in vitro sun protection factor (SPF), UVA protection factor (PF-UVA), and photostability. All the Nps were in the nanometric scale. PI showed monodisperse systems. ZP became more negative as the Np were lyophilized and were added to the formulations. EE varied from 84 to 90%. The SAC went from 44 to 65 microg of sunscreen agents by milligram of Np. The process yield went from 60 to 76%. Nps were predominantly spherical and elliptical forms. The addition of Np diminished the release of the SA. The SPF increased with Np presence and helped to maintain the PF-UVA after irradiation. The HET-CAM assay evaluated the formulation as slightly irritant, CAM-TBS and RBC tests as non irritant, and the Draize test as moderately irritant.

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In vitro quantitative determination of ophthalmic irritancy by the chorioallantoic membrane test with trypan blue staining as alternative to eye irritation test

Cited by 23 publications

References 11 publications

Alternatives toIn VivoDraize Rabbit Eye and Skin Irritation Tests with a Focus on 3D Reconstructed Human Cornea-Like Epithelium and Epidermis Models

Alternatives toIn VivoDraize Rabbit Eye and Skin Irritation Tests with a Focus on 3D Reconstructed Human Cornea-Like Epithelium and Epidermis Models

In vivo and in vitro evaluation of octyl methoxycinnamate liposomes

Characterization and Evaluation of Poly(ε-caprolactone) Nanoparticles Containing 2-Ethylhexyl-p-Methoxycinnamate, Octocrylene, and Benzophenone-3 in Anti-Solar Preparations

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