Abstract: Objective: To explore the method for preparation of decellularized scaffold (DS) from submandibular glands (SMGs) of rats and provide scaffolds for tissue engineering regeneration of salivary glands. Methods: Perfusion system of SMGs was built by ductal and venous cannula. Decellularized SMGs scaffolds were produced by freeze-thaw cycle and sequence perfusion with SDS, Triton X-100, and DNase I through the cannula of vein. HE staining and SEM were performed to observe the general and microstructure of the DS, while Masson staining and immunofluorescence staining were performed to assess extracellular matrix retention. At the same time, the casts of duct and vein for SMGs and DS were compared to analyze the retention of ductal and venous system. Finally, the retention of collagen and DNA was detected by quantitative analysis. Results: Perfusion system was constructed by ductal and venous cannula and DS was produced successfully by perfusion method. DS held the original structure and kept retention of collagen type Ⅰ, Ⅳ, fibronectin, and laminin in extracellular matrix. DNA quantification showed that less than 50 μg/g DNA remained on dry tissue. The casts suggested relatively intact ductal and venous system after decellularization. Conclusion: Decellularized salivary glands scaffolds can be successfully prepared by perfusion method and it will provide scaffolds for tissue engineering construction of salivary glands.

Key words: submandibular glands, decellularized scaffolds, tissue engineering regeneration