A Decellularized Matrix Hydrogel Derived from Human Dental Pulp Promotes Dental Pulp Stem Cells Proliferation, Migration, and Induced Multidirectional Differentiation in vitro.

ABSTRACT Introduction Dental pulp is a major composition in pulp-dentin complex, which serves as protective system against dental trauma/infection. A functional dental pulp regeneration is highly desirable after pulpitis or pulp necrosis. However, endodontic regeneration has remained challenging for decades, due to the deconstructive microenvironment and lack of functional cells within root canal system. The present study developed a decellularized matrix hydrogel derived from human dental pulp (hDDPM-G), which might serve as a growth-permissive microenvironment for dental pulp regeneration. Methods Human dental pulps extracted from healthy wisdom teeth were decellularized and digested, then underwent sol-gel transition to form hDDPM-G. The protein compositions were identified by proteomic analysis. Human dental pulp stem cells (hDPSCs) were seeded on hDDPM-G coated surfaces and evaluated by immunofluorescence staining, Transwell migration, and CCK-8 assays. Induced hDPSCs differentiation was examined in vitro and characterized by immunostaining, western blotting, and RT-PCR. Results Complete decellularization was implemented. Protein contents found in hDDPM were identified to contribute in promoting cell proliferation, migration, and regulation of stem cell differentiation. The hDDPM-G coated surfaces promoted hDPSCs adhesion, migration, and proliferation. Furthermore, hDDPM-G coatings facilitated odontoblast-like, neural-like, and angiogenic differentiation of the seeded hDPSCs after cultured in induction media for 14 days. Conclusions This study demonstrated that hDDPM-G effectively contributed in promoting hDPSCs proliferation, migration, and induced multidirectional differentiation. Considering the injectability and gelation at body temperature, hDDPM-G may hold translational potential for endodontic regeneration.