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Animal experimental models are ruled out by respecting the 3Rs (Replacement, Reduction, Refinement) rules which governed the experimental research for decades with an increased tendency to minimize as much as it is possible any pain suffering or distress that the animals might feel. The chick embryo chorioallantoic membrane (CAM) model is an alternative to other experimental models due to its superior properties compared to other animal models. The CAM is painless by itself due to the lack of innervation and has no immune cells till the 11th day of incubation. Thus, it is extensively used for implanting malignant tumors and assessing them in relation to their metastatic and angiogenic potential. Also, various biomaterials from collagen to hard scaffolds can be implanted on the CAM surface and analyzed mainly related to their property of inducing inflammation. Dental research often uses mouse or rabbit models for experimental purposes. Different surgical techniques from experimentally induced periodontal disease to experimental dental implants may cause pain and suffering to animals. Due to all these arguments, the CAM model is a quick, cheap, and reliable alternative to other animal experimental models used in dental research. Despite its usefulness as an experimental model for different applications, ranging from inflammation studies to cancer research, the CAM model is insufficiently used in dental research. Currently, about 135 studies pertaining to this issue are available in PubMed, the majority of which focus on the reactivity of CAM vessels to various materials employed in dentistry. Limited data exist about the capacity of the CAM to promote osteogenic differentiation of dental stem cells or to enhance biomaterial integration into novel tissue architectures. The present review critically analyzed the use of the CAM model as an experimental tool in dental research. We selected from PubMed all the papers having as topic the CAM in dentistry by searching based on the following keywords: " chorioallantoic membrane, dentistry" or "chorioallantoic membrane, dental ". We focused on discussing the benefits and limitations of the CAM model in dental studies and its prospective role as a preclinical instrument for the assessment of dental tissues, biomaterials, or different dentistry-related substances prior to their use for various purposes in dental clinical practice. ​​​​​​The impact of the CAM model-derived preclinical findings on clinical practice will be also stated by mentioning "pros and cons" arguments. The last part of the present paper reviewed the perspective of CAM assay used in combination with other experimental techniques such as tooth organoids and also the strengths and weaknesses of other species CAM assays recently developed in ostrich and Nile crocodile CAMs., Competing Interests: Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Caplar et al.)

Background/aim: Biomaterials are essential in modern medicine, both for patients and research. Their ability to acquire and maintain functional vascularization is currently debated. The aim of this study was to evaluate the vascularization induced by two collagen-based scaffolds (with 2D and 3D structures) and one non-collagen scaffold implanted on the chick embryo chorioallantoic membrane (CAM)., Materials and Methods: Classical stereomicroscopic image vascular assessment was enhanced with the IKOSA software by using two applications: the CAM assay and the Network Formation Assay, evaluating the vessel branching potential, vascular area, as well as tube length and thickness., Results: Both collagen-based scaffolds induced non-inflammatory angiogenesis, but the non-collagen scaffold induced a massive inflammation followed by inflammatory-related angiogenesis. Vessels branching points/Region of Interest (Px^2) and Vessel branching points/Vessel total area (Px^2), increased exponentially until day 5 of the experiment certifying a sustained and continuous angiogenic process induced by 3D collagen scaffolds., Conclusion: Collagen-based scaffolds may be more suitable for neovascularization compared to non-collagen scaffolds. The present study demonstrates the potential of the CAM model in combination with AI-based software for the evaluation of vascularization in biomaterials. This approach could help to reduce and replace animal experimentation in the pre-screening of biomaterials., (Copyright © 2024, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

A rare, uncommon disorder called PHACE(S) (P-posterior fossa anomalies, H-hemangioma, A-arterial anomalies, C-cardiac anomalies, E-eye anomalies, and S-sternal cleft) of unknown etiology was rarely reported. Children are susceptible to developing PHACE(S) syndrome from the moment they are born. It may be challenging for a physician to appropriately diagnose and treat children with PHACE due to the multifaceted nature of the disease and the extensive range of consequences that may be associated with it. A one-month-old newborn girl was admitted to hospital with extensive, multiple facial infantile hemangiomas, ulceration of the lower lip hemangioma-like lesion, cardiovascular, sternal, and neurological concomitant malformations. Five days following the initial application of the medication, systemic treatment with propranolol and topical treatment with silver sulfadiazine produced their first noticeable benefits. The lip ulceration was mostly healed and facial hemangioma started to regress. The regression continued under therapy and this effect persists for 6 months since Propranolol therapy ended. No cardiovascular or neurological clinical events have been registered during follow-up. The present case has three peculiarities: (1) high number of facial hemangiomas; (2) presence of subependymal cyst not yet reported in the literature associated with PHACE syndrome; and (3) lack of cardiovascular events during therapy knowing that these events frequently appear in PHACE syndrome patients.

Background: Overexpression of chloride intracellular channel protein 1 (CLIC1) in tumor cells has been confirmed, but it has received less attention in the tumor blood vessel endothelium. Aim: The assessment of CLIC1 expression in ccRCC tumor blood vessels and its relationship with TNM parameters and tumor cell CLIC1 expression. Methods: CLIC1 immunostaining in ccRCC was evaluated in 50 cases in both malignant cells and tumor blood vessels (CLIC1 microvessel density-CLIC1-MVD) and was correlated with TNM staging parameters. Results: CLIC1-MVD was observed in approximately 65% of cases, and CLIC1 co-localization in both tumor and endothelial cells was observed in 59% of cases. ccRCC was classified into four groups (Classes 0−3) based on the percentage of positive tumor cells, with each group including sub-groups defined by CLIC1 expression in the endothelium. Class 3 (60−100% positive tumor cells) had the highest CLIC1-MVD, with an impact on T and M parameters (p value = 0.007 for T, and p value = 0.006 for M). For cases with CLIC1 intracellular translocation, there was a strong correlation between CLIC1-MVD and M (p value < 0.001). Conclusions: Co-expression of ccRCC tumor and endothelial cells promotes tumor progression and metastasis and should be investigated further as a potential therapeutic target for ccRCC and other human malignancies.