Evolving applications of the egg: chorioallantoic membrane assay and<i>ex vivo</i>organotypic culture of materials for bone tissue engineering

Marshall, Karen; Kanczler, Janos M.; Oreffo, Richard O.C.

doi:10.1177/2041731420942734

Cited by 22 publications

(26 citation statements)

References 161 publications

(325 reference statements)

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“…The limiting membrane represents the innermost, very thin, dense structure (non-fibrous) of the ESM, which surrounds the egg white. 14,[20][21][22][23][24][25] ^Bellairs and Boyde 21 reported that after staining the eggshell membrane with fluorescein isothiocyanate (FITC), the limiting membrane appears as particles that fill the spaces between the inner membrane fibres, several microns outward from the level at which the inner membrane fibres first appears.…”

Section: Introductionmentioning

confidence: 99%

The eggshell membrane: A potential biomaterial for corneal wound healing

Mensah

Kim

et al. 2021

J Biomater Appl

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The eggshell membrane (ESM) is an abundant resource with innate complex structure and composition provided by nature. With at least 60 million tonnes of hen eggs produced globally per annum, utilisation of this waste resource is highly attractive in positively impacting sustainability worldwide. Given the morphology and mechanical properties of this membrane, it has great potential as a biomaterials for wound dressing. However, to date, no studies have demonstrated nor reported this application. As such, the objective of this investigation was to identify and optimise a reproducible extraction protocol of the ESM and to assess the physical, chemical, mechanical and biological properties of the substrate with a view to use as a wound dressing. ESM samples were isolated by either manual peeling (ESM-strip) or via extraction using acetic acid [ESM-A0.5] or ethylenediaminetetraacetic acid, EDTA [ESM-E0.9]. Energy dispersive X-ray spectroscopy (EDS) confirmed that there were no traces of calcium residues from the extraction process. Fourier transform infrared (FTIR) spectroscopy revealed that the extraction method (acetic acid and EDTA) did not alter the chemical structures of the ESM and also clarified the composition of the fibrous proteins of the ESM. Scanning electron microscopy (SEM) analyses revealed a three-layer composite structure of the ESM: an inner layer as continuous, dense and non-fibrous (limiting membrane), a middle layer with a network of fibres (inner shell membrane) and the outer layer (outer shell membrane) of larger fibres. Material properties including optical transparency, porosity, fluid absorption/uptake, thermal stability, mechanical profiling of the ESM samples were performed and demonstrated suitable profiles for translational applications. Biological in vitro studies using SV40 immortalised corneal epithelial cells (ihCEC) and corneal mesenchymal stromal cells (C-MSC) demonstrated excellent biocompatibility. Taken together, these results document the development of a novel sustainable biomaterial that may be used for ophthalmic wounds and/or other biomedical therapies.

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Section: Introductionmentioning

confidence: 99%

The eggshell membrane: A potential biomaterial for corneal wound healing

Mensah

Kim

et al. 2021

J Biomater Appl

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“…We investigated the antiangiogenic potential of the two compounds by means of CAM assay which was introduced in the early 20th century as an alternative to in vivo experiments due to ethical considerations [ 56 ]. The CAM assay reacts through an inflammatory response to external stimuli in a similar manner with mammalian models thus providing the possibility to extrapolate the results in terms of biocompatibility; in addition, due to native immunodeficiency, the CAM allows the growth of malignant xenografts [ 57 ], which makes it a valuable model for tumor development and testing of anticancer drugs. The procedure is simple, fast, relatively not expensive [ 58 ], and may provide the identification of early biological responses that might me overlooked in animal experiments [ 57 ]; in addition, the method allows treatment testing on patient-derived tumor models within the development of precision medicine [ 59 ].…”

Section: Resultsmentioning

confidence: 99%

3-Pyridinylidene Derivatives of Chemically Modified Lupane and Ursane Triterpenes as Promising Anticancer Agents by Targeting Apoptosis

Казакова

Èoica

Бабаев

et al. 2021

IJMS

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Cancer persists as a global challenge due to the extent to which conventional anticancer therapies pose high risks counterbalanced with their therapeutic benefit. Naturally occurring substances stand as an important safer alternative source for anticancer drug development. In the current study, a series of modified lupane and ursane derivatives was subjected to in vitro screening on the NCI-60 cancer cell line panel. Compounds 6 and 7 have been identified as highly active with GI50 values ranging from 0.03 µM to 5.9 µM (compound 6) and 0.18–1.53 µM (compound 7). Thus, these two compounds were further assessed in detail in order to identify a possible antiproliferative mechanism of action. DAPI (4′,6-diamidino-2-phenylindole) staining revealed that both compounds induced nuclei condensation and overall cell morphological changes consistent with apoptotic cell death. rtPCR analysis showed that both compounds induced upregulation of proapoptotic Bak and Bad genes while downregulating Bcl-XL and Bcl-2 antiapoptotic genes. Molecular docking analysis revealed that both compounds exhibited high scores for Bcl-XL inhibition, while compound 7 showed higher in silico Bcl-XL inhibition potential as compared to the native inhibitor ATB-737, suggesting that compounds may induce apoptotic cell death through targeted antiapoptotic protein inhibition, as well.

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“…The ex ovo variant of the classical in ovo CAM assay offers several unique advantages for drug testing. However, it has been reported by several authors that the ex ovo CAM assay is associated with worse survival rates ( 17 , 28 ). This is in line with our previous experiments during which the survival of embryos in ex ovo CAM assay is between 50% and 70%.…”

Section: Discussionmentioning

confidence: 99%

Extending the Applicability of In Ovo and Ex Ovo Chicken Chorioallantoic Membrane Assays to Study Cytostatic Activity in Neuroblastoma Cells

et al. 2021

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PurposeThe chick chorioallantoic membrane (CAM) assay can provide an alternative versatile, cost-effective, and ethically less controversial in vivo model for reliable screening of drugs. In the presented work, we demonstrate that CAM assay (in ovo and ex ovo) can be simply employed to delineate the effects of cisplatin (CDDP) and ellipticine (Elli) on neuroblastoma (Nbl) cells in terms of their growth and metastatic potential.MethodsThe Nbl UKF-NB-4 cell line was established from recurrent bone marrow metastases of high-risk Nbl (stage IV, MYCN amplification, 7q21 gain). Ex ovo and in ovo CAM assays were optimized to evaluate the antimetastatic activity of CDDP and Elli. Immunohistochemistry, qRT-PCR, and DNA isolation were performed.ResultsEx ovo CAM assay was employed to study whether CDDP and Elli exhibit any inhibitory effects on growth of Nbl xenograft in ex ovo CAM assay. Under the optimal conditions, Elli and CDDP exhibited significant inhibition of the size of the primary tumor. To study the efficiency of CDDP and Elli to inhibit primary Nbl tumor growth, intravasation, and extravasation in the organs, we adapted the in ovo CAM assay protocol. In in ovo CAM assay, both studied compounds (CDDP and Elli) exhibited significant (p < 0.001) inhibitory activity against extravasation to all investigated organs including distal CAM.ConclusionsTaken together, CAM assay could be a helpful and highly efficient in vivo approach for high-throughput screening of libraries of compounds with expected anticancer activities.

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Evolving applications of the egg: chorioallantoic membrane assay andex vivoorganotypic culture of materials for bone tissue engineering

Cited by 22 publications

References 161 publications

The eggshell membrane: A potential biomaterial for corneal wound healing

The eggshell membrane: A potential biomaterial for corneal wound healing

3-Pyridinylidene Derivatives of Chemically Modified Lupane and Ursane Triterpenes as Promising Anticancer Agents by Targeting Apoptosis

Extending the Applicability of In Ovo and Ex Ovo Chicken Chorioallantoic Membrane Assays to Study Cytostatic Activity in Neuroblastoma Cells

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