Rapid SDS/trypsin decellularization of rat submandibular gland yields an ECM scaffold supporting salivary gland tissue engineering

Introduction Salivary gland hypofunction caused by radiation, Sjögren’s syndrome, or other insults leads to xerostomia and currently lacks effective regenerative treatments. Decellularized organ-specific extracellular matrix (ECM) scaffolds offer a promising strategy toward gland restoration by recapitulating the native microenvironment. Methods In this study, we developed a rapid decellularization protocol for rat submandibular glands (SMGs) to create a biomimetic ECM scaffold for salivary gland tissue engineering. Following freeze–thaw pretreatment, detergent and enzyme incubations totaled 4 h and the overall detergent/enzyme processing time, including intermediate PBS washes, was approximately 5 h. The resulting scaffold was characterized by histology, immunostaining, biochemical analysis, and proteomic analysis. Recellularization of the decellularized SMG (dSMG) was performed via intraductal injection of human submandibular gland mesenchymal stem cells (hMSCs) or human submandibular gland stem cells (hSMG-SCs), followed by in vitro culture. In addition, hSMG-SC-seeded dSMG scaffolds were implanted subcutaneously in immunodeficient mice for in vivo evaluation. Results The protocol achieved effective decellularization, reducing residual DNA to