Study of the functional relationships between the buccinator muscle and the connective tissue of the cheek in humans.

BACKGROUND: The buccinator muscle derives from the mesenchyme of the second pharyngeal arch. In adults, it has a quadrilateral shape, occupying the deepest part of the cheek region. Its function is complex, being active during swallowing, chewing, and sucking. To our knowledge, there are no studies that have specifically analyzed the relationship of the buccinator muscle fibers and neighboring connective tissue of the cheek in humans, neither during development nor in adults. Such relationships are fundamental to understand its function. Thus, in this study the relations of the buccinator muscle with associated connective tissue were investigated. METHODS: The buccinator muscle region was investigated bilaterally in 41 human specimens of 8-17 weeks of development. Moreover, four complete adult tissue blocks from human cadavers (including mucosa and skin) were obtained from the cheek region (between the anterior border of the masseter muscle and the nasolabial fold). All samples were processed with standard histological techniques. In addition, subsets of sections were stained with picrosirius red (PSR). Furthermore, immunoreactivity against type I and III collagen was also studied in adult tissues. RESULTS: The buccinator muscle showed direct relationships with its connective tissue from 8 to 17 weeks of development. Collagen fibers were arranged in septa from the submucosa to the skin through the muscle. These septa were positive for type I collagen and presented elastic fibers. Fibrous septa that were positive for type III collagen were arranged from the lateral side of the muscle to the skin. CONCLUSIONS: The intimate relationship between buccinator muscle fibers and cheek connective tissue may explain the complex functions of this muscle.

Interactions between TGF-beta1 and TGF-beta3 and their role in medial edge epithelium cell death and palatal fusion in vitro.

In recent decades, studies have shown that both TGF-beta(1) and TGF-beta(3) play an important role in the induction of medial edge epithelium (MEE) cell death and palatal fusion. Many of these experiments involved the addition or blockage of one of these growth factors in wild-type (WT) mouse palate cultures, where both TGF-beta(1) and TGF-beta(3) are present. Few studies have addressed the existence of interactions between TGF-beta(1) and TGF-beta(3), which could modify their individual roles in MEE cell death during palatal fusion. We carried out several experiments to test this possibility, and to investigate how this could influence TGF-beta(1) and TGF-beta(3) actions on MEE cell death and palatal shelf fusion. We double-immunolabelled developing mouse palates with anti-TGF-beta(1) or anti-TGF-beta(3) antibodies and TUNEL, added rhTGF-beta(1) or rhTGF-beta(3) or blocked the TGF-beta(1) and TGF-beta(3) action at different concentrations to WT or Tgf-beta(3) null mutant palate cultures, performed in situ hybridizations with Tgf-beta(1) or Tgf-beta(3) riboprobes, and measured the presence of TUNEL-positive midline epithelial seam (MES) cells and MES disappearance (palatal shelf fusion) in the different in vitro conditions. By combining all these experiments, we demonstrate great interaction between TGF-beta(1) and TGF-beta(3) in the developing palate and confirm that TGF-beta(3) has a more active role in MES cell death than TGF-beta(1), although both are major inductors of MES disappearance. Finally, the co-localization of TGF-beta(1), but not TGF-beta(3), with TUNEL in the MES allows us to suggest a possible role for TGF-beta(1) in MES apoptotic clearance.