The studies of lymphatic architecture and distribution are increasing in importance in the investigation of the routes of tumor metastasis, treatment of edema and the development of lymphaticovenular anastomosis. In this study, lymphatic architecture and distribution beneath the buccal mucosa of mice were investigated using enzymatic histochemical staining and analysis of the three-dimensional lymphatic architecture using a new 3D reconstruction method. Using the obtained histological image, buccal mucosa was vertically divided into three areas including the upper area from corresponding buccal mucosa to occlusal plane (U), corresponding buccal mucosa to occlusal plane (O) and lower area from corresponding buccal mucosa to occlusal plane (L) areas. Anteroposteriorly, buccal mucosa was divided into four areas comprising the corresponding buccal mucosa to incisor (I), corresponding buccal mucosa to edentulous area (E), corresponding buccal mucosa to molar (M) and fauces area of buccal mucosa (F) areas. Network formation of the lymphatic vessels was observed subepithelially at the buccal mucosa. The networks of the lymphatic vessels were finer than the blood vessels. The precollecting lymphatics heading to the suprafascial collecting lymphatics right below the mucosa was observed on O area. Entering short connective tissue papillae, blind-ends were observed most on LM and OM areas followed by LE and OF areas. It is suggested that the thickness of cornified layers in mucosal epithelium and the entire thickness of the epithelial layers has an effect on drug penetration. In addition to that, UM area is considered to be an effective area to place drugs that require instantaneous effects in terms of penetration. Our results also suggested that OM area is an effective place for drugs with high molecular weight.
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