An innovative method to achieve functional dental pulp regeneration with the complex scaffold of atelocollagen gelatin sponge and treated dentin matrix

牙髓疾病严重影响患者的生活质量，造成巨大的经济负担。功能性牙髓再生作为根管治疗的一种替代方法，已经引起了人们的极大关注。本研究旨在开发一种创新的方法，以实现新的血管和感觉神经的功能性牙髓再生，为未来的临床治疗。组织再生需要合适的生物材料支架、生物活性分子和牙髓干细胞等关键因素。由于去端胶原/明胶（ACG）海绵的主要成分与牙髓组织的胶原蛋白相同，因此其成功制备具有很大的潜力。ACG海绵也可以被修饰或掺入生物活性试剂作为药物递送系统。真空热处理没有引入任何有毒的交联剂，有效地提高了ACG海绵的结构稳定性。此外，还成功制备了ACG/β-TCP/Mg 2+复合海绵，并显示出相似的多孔结构。SEM和XPS分析表明，活性β-TCP和Mg 2+被完全引入到ACG海绵中。从4周龄SD大鼠切牙中分离培养牙髓干细胞。确定了牙髓干细胞体外增殖和分化的最适Mg 2+浓度。ACG/β-TCP/Mg 2+复合海绵不影响细胞在海绵表面的生长和增殖。这表明ACG海绵结合生物活性分子为组织工程提供了可能。

Dental pulp diseases severely compromise patients' quality of life and cause huge economic burden. Functional dental pulp regeneration has gained enormous attention as an alternative to root canal treatment. This study aimed to develop an innovative method to achieve functional dental pulp regeneration with new blood vessels and sensory nerves for future clinical treatments. Generally, key elements including suitable biomaterial scaffolds, bioactive molecules and dental pulp stem cells are needed for tissue regeneration. The successful fabrication of atelocollagen/gelatin (ACG) sponge holds great potential as it shares the main component of collagen with dental pulp tissue. ACG sponge could also be modified or incorporated with bioactive reagents as a drug delivery system. Heat treatment in the vacuum, which did not introduce any toxic crosslinking reagent, effectively improved the structural stability of ACG sponge. Moreover, ACG/β-TCP/Mg2+ composite sponge had been successfully fabricated and showed similar porous microstructures. Bioactive β-TCP and Mg2+ were shown to be fully incorporated into ACG sponge as revealed by SEM and XPS analyses. Dental pulp stem cells were isolated and cultured from incisors of 4-week-old SD rats. The optimal concentration of Mg2+ for dental pulp stem cells proliferation and differentiation was confirmed in vitro. ACG/β-TCP/Mg2+ composite sponge did not compromise cell growth and proliferation on the sponge surface. These indicate that ACG sponge combined with bioactive molecules may pose the possibility for tissue engineering.