Tissue engineering strategies for construction of artificial salivary glands based on decellularized scaffolds of...

Salivary gland (SG) hypofunction is a debilitating condition with no curative treatments. Tissue engineering based on decellularized scaffolds represents a promising regenerative strategy; however, its application to SGs has been limited by inadequate decellularization techniques and scarce cell sources. Here, we developed a novel perfusion-based decellularization method via the ductal and venous systems to generate rat submandibular gland scaffolds. This approach efficiently removed cellular components while optimally preserving the native three-dimensional architecture, key extracellular matrix (ECM) components, and the integrity of ductal and vascular networks. We then constructed artificial SGs by recellularizing these biomimetic scaffolds through the ductal route with salivary gland cells (SGCs), adipose-derived stromal cells (ADSCs), or SGC-pre-induced ADSCs. In vitro perfusion culture revealed that all cell sources, including transdifferentiated ADSCs, supported the formation of acinar-like structures and expressed the functional marker α-amylase (α-AMY), with peak performance observed at day 5. Upon in vivo transplantation, the tissue-engineered glands maintained partial α-AMY expression for up to 5 days, demonstrating short-term survival and function. However, graft longevity was limited beyond 7 days, primarily due to insufficient vascularization. Our study establishes a robust perfusion-decellularized scaffold as a superior biomaterial platform and validates ADSCs as a clinically viable cell source for SG regeneration, providing a foundational strategy for engineering functional salivary glands. This advancement is expected to accelerate the research and development in the field of tissue engineering for salivary gland regeneration.