The Chick Embryo Chorioallantoic Membrane (CAM) Assay: A Novel Experimental Model in Dental Research

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.