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一种替代根管治疗的方法已经引起了广泛的关注。本研究旨在发展一种具有新生血管和感觉神经功能的牙髓再生方法，为今后的临床治疗提供参考。一般来说，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要素。由于ACG海绵与牙髓组织共享胶原蛋白的主要成分，因此ACG海绵的成功制备具有很大的潜力。ACG海绵也可以修饰或与生物活性试剂结合作为药物输送系统。真空热处理，不引入任何有毒交联试剂，有效提高了ACG海绵的结构稳定性。成功制备了ACG/β-TCP/Mg2+复合海绵，并表现出相似的多孔微观结构。通过扫描电镜和XPS分析发现，具有生物活性的β-TCP和Mg2+完全融入到ACG海绵中。从4周龄SD大鼠门牙中分离培养牙髓干细胞。确定了Mg2+对牙髓干细胞体外增殖分化的最佳浓度。ACG/β-TCP/Mg2+复合海绵不影响海绵表面细胞的生长和增殖。这表明ACG海绵与生物活性分子的结合有可能用于组织工程。

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.