Abstract
Regenerative muscles are required for swallowing and mastication, and are important for functional recovery from diseases involving oral muscular defects. Therefore, we generated three-layer hybrid sheets, similar to oral mucosal structures containing submucosal muscles, using rabbit oral mucosa epithelial, mesenchymal, and myoblastic progenitor cells, and examined the structural proteins. Each cell type was obtained from rabbit oral mucosa using enzymatic digestion. Isolated mesenchymal and myoblastic cells were multi-differentiated into osteoblasts, adipocytes, and chondrocytes or myotubes. Isolated epithelial cells were cultured on collagen gels containing isolated mesenchymal cells for 2 weeks, and these epithelial–mesenchymal cell sheets were laminated onto myoblastic cell sheets. The engineered hybrid sheets were multi-stratified in the epithelial and myoblastic layers in a time-dependent manner, expressing intermediate cytoskeletal filament proteins of epithelium and muscle. Hybrid sheets also expressed extracellular matrix basement membrane proteins. Immature cell markers for epithelial and myoblastic cells were observed continuously in hybrid sheet cultures. We established engineered three-dimensional rabbit oral mucosa hybrid sheets containing each immature cell type in vitro.
Highlights
Mucous membranes line the oral cavity, pharynx, and esophagus, which are involved in mastication and swallowing
Isolation of mesenchymal cells and myoblasts from oral mucosa and their analysis Cells collected from rabbit oral mucosa mesenchymal tissues were successfully amplified from single cells by methylcellulose culture
Oil redpositive lipid droplets were observed in adipogenic-induced rabbit oral mucosa mesenchymal tissues (rOMMCs), and cartilage mucin staining Safranin O-positive cells were observed in chondrogenic-induced rOMMCs (Figures 2b and 2c)
Summary
Mucous membranes line the oral cavity, pharynx, and esophagus, which are involved in mastication and swallowing. Beneath the mucosal epithelium is a layer of muscles that includes the buccinator muscles and the pharyngeal constrictors, among others. This continuous muscle layer has a crucial role in oral cavity and pharynx function. Antebrachial flaps and rectus abdominis flaps have often been used for reconstruction following the removal of malignant tumors from the oral cavity, such as tongue cancers or pharyngeal cancer, and have enabled recovery of the minimally required shape and function of the oral cavity.[1,2,3] recovery of muscle function required for swallowing and chewing is difficult with conventional methods employing grafted tissue.
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