Study protocol on the impact of postnatal maternal separation stress on dental enamel formation in a murine experimental model

Dental development is a complex process influenced by genetic and environmental factors. Dental enamel, primarily composed of hydroxyapatite, is formed through complex cellular and biochemical mechanisms. Although this is a stable process, genetic, nutritional, and environmental factores can lead to developmental defects such as hypomineralization and hypoplasia. Molar incisor hypomineralization is a type of hypomineralization that represents a public health challenge. Its etiology is not yet fully understood, but factors such as hypoxia, medication exposure, adverse events in early childhood, and genetic influences are considered. This study protocol aims to investigate whether postnatal adverse events can impact amelogenesis, exploring the role of stress in the etiology of dental enamel defects. Specific objectives include evaluating enamel structure and mechanical properties by comparing the offspring of rats exposed to postnatal maternal separation with control animals (non-exposed). Additionally, we will evaluate weight, length, survival assessment, and developmental milestones between the groups. Macrophotographic analysis, microtomography, microhardness testing, and electron microscopy will enable a detailed assessment of enamel morphology and its mechanical properties. Histological and molecular analyses-such as immunohistochemistry, indirect immunofluorescence, and in situ zymography-will be performed to evaluate possible changes in proteins and enzymes that are essential for proper enamel biomineralization.