The treatment and care of the animals was done in accordance with the regulations set forth by the Cairo Agriculture for Experimental Animals’ Animal Ethics Committee, Cairo, Egypt, Approval No (113-10-22). According to the Declaration of Helsinki and its Guiding Principles for the Care and Use of Animals (NIH Publication No. 85-23, 1985 revision), researchers adhered to certain rules. The experimental rats were housed in cages in the laboratory with free access to food and water. In order to reduce suffering, proper care was given to the animals. Prior to the experiment, the animals were acclimated for at least 14 days under typical circumstances of 25 °C temperature and 55.5% relative humidity with a 12-hour cycle of light and darkness. The experiment involved 48 adult rats, which were divided into 16 groups of 3 rats each. Every group received treatment using a formulation chosen based on the study design. The following steps were taken to construct the in-vivo periodontitis model. Chloral hydrate 10% was injected intraperitoneally to anesthetize the rats. After that, they were secured in the supine posture, and a homemade mouth expander was used to open the mouth. A gingival separator was used to separate the gums and totally expose the bilateral maxillary first molars. Each rat in the group had both of their maxillary first teeth tied together for a period of 2 weeks using an orthodontic steel wire (0.2 mm in diameter). By placing the ligation wires in the gingival sulcus, the ligation did not interfere with the rats’ ability to feed or harm their buccal mucosa. In other groups, the periodontitis model was developed in a single day. The buccal, lingual, and mesial gingiva of the maxillary molars, as well as the adjacent tooth space between the maxillary molars, were then injected with variously produced CrO-Tur-SNEDDS (60 µl for each tooth).
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